培土生金中药对脾虚哮喘大鼠肺、回肠组织神经肽水平的影响
|
摘要
目的:本实验通过研究脾虚哮喘及哮喘大鼠肺、回肠组织神经肽及受体表达水平的改变,探讨脾虚哮喘及哮喘动物机体内的神经内分泌免疫调节失衡状态;通过对脾虚哮喘及哮喘大鼠肺、回肠组织神经肽及受体表达水平改变的比较研究,探讨脾虚对哮喘的影响,为“脾虚为哮喘之关键病机”的论点提供新的依据,以进一步验证和深化哮喘的中医病机理论;通过研究培土生金中药对脾虚哮喘及哮喘大鼠肺、回肠组织神经肽及受体表达水平的影响,探讨培土生金法治疗哮喘的作用机制;通过对脾虚哮喘及哮喘大鼠肺、回肠组织神经肽及受体表达水平相关性的研究,探讨培土生金理论的现代科学内涵和肺脾二脏的密切关系。
材料与方法:以中医理论为指导,复合利用中医脾虚动物模型和西医哮喘动物模型的造模方法,建立大鼠脾虚哮喘病证结合的模型。脾虚动物造模采用饮食不节加疲劳过度的方法。造模结束后,比较两模型组动物造模前后体重、肛温变化情况,以脾虚模型评估标准评估两模型组动物症状。以动物定量指标的变化有统计学意义、症状符合脾虚模型评估标准来判定脾虚动物模型建立成功。哮喘动物造模采用以卵蛋白为致敏原,氢氧化铝、百日咳灭活杆菌为免疫佐剂,腹腔注射致敏后以卵蛋白液雾化吸入激发建立动物哮喘模型。以大鼠出现呼吸加深加快,点头运动明显,腹肌强烈收缩,偶可闻及痰鸣音等类似人类哮喘的症状为造模成功。造模成功后,采用培土生金中药进行干预,共12天。干预结束后,采用血液白细胞及嗜酸粒细胞绝对计数方法测定大鼠末梢血及支气管肺泡灌洗液(BALF)中白细胞(WBC)、嗜酸性粒细胞(EOS)计数;采用双抗体夹心酶联免疫法(ELISA)测定大鼠血清及BALF中自介素-4(IL-4)、干扰素-γ(IFN-γ)的浓度;采用免疫组化方法半定量分析肺及回肠组织中P物质(SP)、血管活性肠肽(VIP)含量;采用逆转录聚合酶链反应(RT-PCR)方法半定量分析肺组织中SP及其NK1受体mRNA的表达;并应用光镜对肺、回肠组织形态学进行观测。
结果:
1.同对照(C)组比较,疾病加证候(A)组和疾病(B)组大鼠均表现为血中及BALF中白细胞计数、嗜酸性粒细胞计数显著升高(P＜0.01);疾病加证候(A)组和疾病(B)组大鼠均表现为血中及BALF中IL-4的浓度显著升高(P＜0.01),IFN-γ的浓度显著降低(P＜0.01),IFN-γ/IL-4比值显著降低(P＜0.01);疾病加证候(A)组和疾病(B)组大鼠均表现为肺组织中SP及其NK1受体mRNA表达显著增强(P＜0.01);疾病加证候(A)组和疾病(B)组大鼠均表现为肺、回肠组织中SP含量显著增加(P＜0.01),VIP含量显著减少(P＜0.01)。
2.与疾病(B)组比较,疾病加证候(A)组外周血白细胞计数、BALF中白细胞计数、嗜酸性粒细胞计数、血清IL-4浓度、BALF中IL-4浓度、肺组织SP及其NK1受体mRNA表达、肺组织及回肠组织SP含量均显著高于疾病(B)组(P＜0.05);BALF中IFN-γ浓度及IFN-γ/IL-4比值、肺组织及回肠组织VIP含量均显著低于疾病(B)组(P＜0.01或P＜0.05)。
3.与相应的疾病组比较,两中药(A1,B1)组血中及BALF中嗜酸性粒细胞计数显著降低(P＜0.01或P＜0.05)。其中,疾病加证候中药(A1)组BALF中嗜酸性粒细胞计数与对照组比较,已无显著差异(P＞0.05);疾病加证候中药(A1)组血中及BALF中白细胞计数显著降低(P＜0.01),疾病中药(B1)组BALF中白细胞计数显著降低(P＜0.01)。与相应的疾病组比较,两中药组(A1,B1)血中及BALF中IL-4的浓度显著降低(P＜0.01),IFN-γ的浓度显著升高(P＜0.01或P＜0.05),IFN-γ/IL-4比值显著升高(P＜0.01或P＜0.05)。其中,疾病加证候中药(A1)组血中IFN-γ浓度、BALF中IL-4的浓度及BALF中IFN-γ/IL-4比值与对照(C)组比较,已无显著差异(P＞0.05)。与相应的疾病组比较,两中药(A1,B1)组肺、回肠组织中SP含量显著减少(P＜0.01),VIP含量显著增加(P＜0.01),其中,疾病加证候中药(A1)组肺及回肠组织SP含量、疾病中药(B1)组回肠组织SP含量与对照(C)组比较,已无显著差异(P＞0.05)。与相应的疾病组比较,两中药(A1,B1)组肺组织中SP及其NK1受体mRNA表达显著减少(P＜0.01或P＜0.05)。其中,疾病加证候中药(A1)组肺组织SPmRNA表达水平、疾病中药(B1)组肺组织NK1受体mRNA表达水平与对照(C)组比较,已无显著差异(P＞0.05)。
4.各组大鼠血中IFN-γ/IL-4比值与肺组织SP含量间存在着明显的负相关(r=-0.698,P＜0.01);各组大鼠血中IFN-γ/IL-4比值与肺组织VIP含量间存在着明显的正相关(r=0.741,P＜0.01)。各组大鼠血中IFN-γ/IL-4比值与回肠组织SP含量间存在着明显的负相关(r=-0.627,P＜0.01);各组大鼠血中IFN-γ/IL-4比值与回肠组织VIP含量间存在着明显的正相关(r=0.766,P＜0.01)。各组大鼠BALF中IFN-γ/IL-4比值与肺组织SP含量间存在着明显的负相关(r=-0.688,P＜0.01);各组大鼠BALF中IFN-γ/IL-4比值与肺组织VIP含量间存在着明显的正相关(r=0.324,P＜0.01)。各组大鼠BALF中IFN-γ/IL-4比值与回肠组织SP含量间存在着明显的负相关(r=-0.615,P＜0.01);各组大鼠BALF中IFN-γ/IL-4比值与回肠组织VIP含量间存在着明显的正相关(r=0.679,P＜0.01)。各组大鼠肺组织中SP与VIP含量之间存在着明显的负相关(r=-0.678,P＜0.01);各组大鼠回肠组织中SP与VIP含量之间存在着明显的负相关(r=-0.620,P＜0.01)。各组大鼠肺组织与回肠组织中SP含量之间存在着明显的正相关(r=0.602,P＜0.01);各组大鼠肺组织与回肠组织中VIP含量之间存在着明显的正相关(r=0.608,P＜0.01)。
5.病理结果显示:疾病(B)组与疾病加证候(A)组均有不同程度的支气管及肺组织炎性病变,两中药(A1,B1)组的支气管及肺组织的炎性病变均有所减轻;疾病(B)组与疾病加证候(A)组均有不同程度的回肠组织炎性病变,两中药(A1,B1)组的回肠组织炎性病变程度有所减轻,但以疾病加证候治疗(A1)组的减轻程度明显。
结论:
1.哮喘及脾虚哮喘大鼠均存在着明显的气道局部或/和全身炎症性改变;哮喘及脾虚哮喘大鼠均存在着全身及气道局部免疫细胞因子IFN-γ、IL-4表达失衡;哮喘及脾虚哮喘大鼠均存在着肺、回肠组织内神经肽VIP、SP表达水平失衡;哮喘及脾虚哮喘大鼠均存在着肺组织内神经肽SP及其NK1受体mRNA表达水平失衡。
2.脾虚可以加重哮喘机体气道内炎性细胞聚集及外周血炎性细胞聚集,加重哮喘机体肺、回肠组织炎性病变,从而加重哮喘机体的局部及全身炎症反应;脾虚可以加重哮喘机体血清及BALF中免疫细胞因子IFN-γ、IL-4表达失衡,加重哮喘机体肺、回肠组织内神经肽VIP、SP表达水平失衡,加重肺组织内神经肽SP及其NK1受体mRNA表达水平失衡,从而加重哮喘机体的神经内分泌免疫调节失衡,成为哮喘反复发作的内在原因之一。
3.培土生金中药可以减轻模型大鼠气道局部及全身的炎性细胞浸润,下调血中及BALF中白细胞及嗜酸性粒细胞计数;培土生金中药可以调整全身及气道局部免疫细胞因子IFN-γ、IL-4表达失衡状态,上调血清及BALF中IFN-γ浓度,下调IL-4浓度,上调IFN-γ/IL-4比值;培土生金中药可以减轻肺、回肠组织内神经肽VIP、SP及NK1受体表达水平的失衡状态,下调肺组织中SP及其NK1受体mRNA表达水平,下调肺、回肠组织中SP含量表达水平,上调肺、回肠组织中VIP含量表达水平;培土生金中药能够减轻模型大鼠的大、小气道及肺、回肠组织的炎性病变。
4.肺、回肠组织神经肽VIP、SP表达异常及肺组织SP及其NK1受体mRNA表达的异常均与哮喘慢性气道炎症的形成与持续存在明显相关性,与气道局部及全身的炎症状态呈明显相关性,与气道局部及全身的免疫细胞因子IFN-γ、IL-4表达失衡状态亦呈明显相关性。这进一步证实了神经肽VIP、SP在哮喘发病过程中的重要作用,特别是介导哮喘的神经内分泌免疫调节失衡。肺组织与回肠组织的神经肽VIP、SP的表达水平存在着明显的相关性,从神经肽VIP、SP及神经内分泌免疫调节的角度进一步证实了肺脾二脏的密切关系。
OBJECTIVE:We attempted to investigate the imbalanced states of neuro-endocrine-immuno-modulation through study of the changes of neurikins and their receptors of the tissues of the lung and ileum in experimental rates with spleen-deficiency asthma and simple asthma;to investigate the influence of spleen deficiency on asthma by comparison of the changes of expressive levels between the two kinds of rat models,and thus to provide a new evidence for the view points of "the spleen deficiency as the key patho-mechanism".We attempted the effective mechanism of the therapy of "strengthening the earth and promoting the metal(SEPM)" in treatment of asthma through study of the influence of Chinese herbal remedy of SEPM to the expressive levels of neurikins and their receptors of the tissues of the lung and ileum in experimental rates with spleen-deficiency asthma and simple asthma;to investigate the modern connotation of SEPM theory and the close relationship between the lung and the spleen by studying the correlation between the expressive levels of neurikins and their receptors of the tissues of the lung and ileum in experimental rates with spleen-deficiency asthma and simple asthma.
MATERIAL AND METHODS,Taking the theory of traditional Chinese medicine (TCM) as the guide,we set up compound rat models with asthma of spleen deficiency by combination of establishing spleen deficiency animal models in TCM with establishing asthmatic animal models in Western medicine.The spleen models were set up by using the method of improper diet plus physical overstrain.The body weight and temperature of the anus of the animals were compared before and after the model establishment;and the symptoms of the animals were assessed by the standards for evaluation of spleen deficiency models in TCM.The successful animal models were determined by the statistical significance in the changes of their quantitative indexes and their symptomatologic accordance with the standards for evaluation of spleen deficiency.Asthmatic animal models were set up by using ovalbumin as the antigen,and sodium hydroxide and pertusis vaccine as the adjuvant.The asthmatic animals were set up by nebulization following the intraperitoneal injection of valbumin.The successful animal models were evaluated by the appearance of similar symptoms of human asthma in rats such as tachypnea,remarkable motion of nodding,strong construction of abdominal muscle,and occasional pulmonary rale and grunting.The successfully established animal models were given Chinese herbal remedy of SEPM for 12 days.Then the following determinations were conducted:white blood cell count(WBC) and eosinophil count(EOS) in peripheral blood and bronchial alveolar lavage fluid (BALF) by absolute counting;the contents of interleuki-4(IL-4) and interferon-γ(IFN-γ) in BALF by enzyme-linked immunoabsorbent assay(ELISA); the contents of substance P(SP) vasoactive intestinal peptide(VIP) by semi-quantitative analysis of immune-chemical assay;the expressions of SPmRNA and neurokinin 1(NK1) receptor mRNA by reverse transcription-polymerase chain reaction(RT-PCR).
RESULTS:
1.Compared with the control(C) group,the number of WBC and EOS of the peripheral blood and BALF in the compound(A) group and disease(B) group increased(P＜0.01);the content of IL-4 of BALF in group A and B decreased (P＜0.01),and the content of IFN-γof BALF decreased(P＜0.01),and the ratio of IFN-γ/IL-4 was remarkably decreased(P＜0.01).The mRNA expressions of SPmRNA and NK1 receptor mRNA in group A and B enhanced(P＜0.01);the content of SP in group A and B increased(P＜0.01) and the content of VIP obviously decreased(P＜0.01).
2.Compared with group B,the WBC in peripheral blood;the WBC and EOS in BALF;serum IL-4 level,IL-4 level in BALF,the expressions of SPmRNA and NK1 receptor mRNA,and the contents of SP of the lung and ileum tissues in group A all increased remarkably(P＜0.01 or P＜0.05).The level of IFN-γand ratio of IFN-γ/IL-4 in BALF,and the content of VIP in the lung and ileum tissues in group A all decreased remarkably(P＜0.05).
3.Compared with the correspondent disease groups,the EOS in blood and BALF in the two Chinese herbal groups(A1,B1) remarkably decreased(P＜0.01 or P＜0.05).And the EOS of BALF in group A1 had showed no significant difference from that in the control group(P＞0.05).The WBC and EOS of blood and BALF in group A1 obviously decreased(P＜0.01);the WBC of BALF in group B1 obviously decreased(P＜0.01).The levels of IL-4 of blood and BALF in both group A1 and B1 remarkably decreased(P＜0.01),but the levels of IFN-γobviously increased (P＜0.01 or P＜0.05);and the ratio of IFN-γ/IL-4 increased(P＜0.01 or P＜0.05).And the level of IFN-γof blood,the level of IL-4 of BALF,and the ratio of IFN-γ/IL-4 of BALF in group A1 had showed no significant difference from those in group C(P＞0.05).The expressions of SPmRNA and NK1 receptor mRNA in group A1 and B1 were remarkably decreased(P＜0.01 or P＜0.05).And the expressive levels of SPmRNA in group A1 and NK1 receptor mRNA in group B1 had showed no significant difference from those in group C(P＞0.05).The contents of SP in the lung and ileum in both group A1 and B1 obviously decreased(P＜0.01),but the content of VIP remarkably increased(P＜0.01).And the contents of SP in the lung and ileum in group A1,and the content of SP in the ileum in group B1 had showed no significant difference from those in group C(P＞0.05).
4.There was an obviously negative correlation between the ratio of IFN-γ/IL-4 of blood and the content of SP of the lung tissue in each group(r =-0.698,P＜0.01);there was an obviously positive correlation between the ratio of IFN-γ/IL-4 of blood and the content of VIP of the lung tissue(r=0.741, P＜0.01).There was an obviously negative correlation between the ratio of IFN-γ/IL-4 of blood and the content of SP of the ileum tissue in each group (r=-0.627,P＜0.01);there was an obviously positive correlation between the ratio of IFN-γ/IL-4 of blood and the content of VIP of the ileum tissue(r =0.766,P＜0.01).There was an obviously negative correlation between the ratio of IFN-γ/IL-4 of BALF and the content of SP of the lung tissue in each group (r=-0.688,P＜0.01);there was an obviously positive correlation between the ratio of IFN-γ/IL-4 of BALF and the content of VIP of the lung tissue(r= 0.324,P＜0.01).There was an obviously negative correlation between the ratio of IFN-γ/IL-4 of BALF and the content of SP of the ileum tissue in each group (r=-0.615,P＜0.01);there was an obviously positive correlation between the ratio of IFN-γ/IL-4 of BALF and the content of VIP of the ileum tissue(r= 0.679,P＜0.01).There was an obviously negative correlation between the contents of SP and VIP of the lung tissue in each group(r=-0.678,P＜0.01);there was an obviously positive correlation between the contents of SP and VIP of the lung tissue(r=-0.620,P＜0.01).There was an obviously negative correlation between the contents of SP and VIP of the ileum tissue in each group(r=0.602,P＜0.01);there was an obviously positive correlation between the contents of SP and VIP of the ileum tissue(r=0.608,P＜0.01).
5.The pathological results showed that there were bronchial and pulmonary inflammation at different degrees in both group B and A,and the inflammation in both group A1 and B1 had been reduced;there were ileal inflammation at different degrees in both group B and A,and the inflammation in both group A1 and B1 had been reduced,with the inflammation in group A1 was more obvious.
CONCLUSION:
1.There are,in asthmatic and spleen deficiency asthmatic rats,evident airway or/and general inflammation,expressive imbalance of general and airway immune cytokin IFN-γand IL-4,expressive imbalance of neurikins VIP and SP, and expressive imbalance of the SPmRNA and NK1mRNA receptors of the tissues of the lung and ileum,and the imbalance of neuro-endocrine-immuno-modulation.
2.Spleen deficiency can deteriorate the aggregation of inflammatory cells in the peripheral blood and airway of asthmatic rat models,and aggravate the pulmonary and ileal inflammation,and thus exacerbate the general and local inflammatory reaction of the asthmatic body.It can also exacerbate the imbalance of immune cytokins IFN-γand IL-4 in serum and BALF,and aggravate the expressive imbalance of neurikins VIP and SP and the SPmRNA and NK1mRNA receptors of the tissues of the lung and ileum,and thus make the imbalance of neuro-endocrine-immuno-modulation worse,being one of the inner causes for repeated attack of asthma.
3.Chinese remedy of SEPM can reduce the airway and general infiltration of inflammatory cells in rat models,down-regulate WBC and EOS in peripheral blood and BALF.It can also regulate the imbalanced state of general and local airway immune cytokins IFN-γand IL-4,up-regulare the content of IFN-γ, down-regulate the content of IL-4,and up-regulate the ratio of IFN-γ/IL-4. It can relieve the expressive imbalance of neurikins VIP,SP and NK1 receptors of the tissues of the lung and ileum,and down-regulate the expressive levels of SPmRNA and NK1mRNA receptors;down-regulate the expressive levels of SP and up-regulate the expressive levels of VIP in both the lung and ileum tissues. And the Chinese remedy of SEPM can reduce the tissue inflammation of the airways, lungs and ileum of the rat models.
4.The expressive imbalance of neurikins VIP and SP,and the SPmRNA and NK1mRNA receptors of the tissues of the lung and ileum is obviously correlated with the development and persistence of chronic airway inflammation in asthma, with general and local airway inflammatory state,and with the imbalanced state of general and local airway immune cytokins IFN-γand IL-4.This further reveals the important role of neurikins VIP and SP in development of asthma,especially in inducing imbalance of neuro-endocrine-immuno-modulation.There is evident correlation between the expressive levels of VIP and SP in the tissues of the lung and ileum,which further expounds the close relationship between the lung and the spleen from the angle of neurikins VIP and SP and neuro-endocrine-immuno-modulation.
材料与方法:以中医理论为指导,复合利用中医脾虚动物模型和西医哮喘动物模型的造模方法,建立大鼠脾虚哮喘病证结合的模型。脾虚动物造模采用饮食不节加疲劳过度的方法。造模结束后,比较两模型组动物造模前后体重、肛温变化情况,以脾虚模型评估标准评估两模型组动物症状。以动物定量指标的变化有统计学意义、症状符合脾虚模型评估标准来判定脾虚动物模型建立成功。哮喘动物造模采用以卵蛋白为致敏原,氢氧化铝、百日咳灭活杆菌为免疫佐剂,腹腔注射致敏后以卵蛋白液雾化吸入激发建立动物哮喘模型。以大鼠出现呼吸加深加快,点头运动明显,腹肌强烈收缩,偶可闻及痰鸣音等类似人类哮喘的症状为造模成功。造模成功后,采用培土生金中药进行干预,共12天。干预结束后,采用血液白细胞及嗜酸粒细胞绝对计数方法测定大鼠末梢血及支气管肺泡灌洗液(BALF)中白细胞(WBC)、嗜酸性粒细胞(EOS)计数;采用双抗体夹心酶联免疫法(ELISA)测定大鼠血清及BALF中自介素-4(IL-4)、干扰素-γ(IFN-γ)的浓度;采用免疫组化方法半定量分析肺及回肠组织中P物质(SP)、血管活性肠肽(VIP)含量;采用逆转录聚合酶链反应(RT-PCR)方法半定量分析肺组织中SP及其NK1受体mRNA的表达;并应用光镜对肺、回肠组织形态学进行观测。
结果:
1.同对照(C)组比较,疾病加证候(A)组和疾病(B)组大鼠均表现为血中及BALF中白细胞计数、嗜酸性粒细胞计数显著升高(P＜0.01);疾病加证候(A)组和疾病(B)组大鼠均表现为血中及BALF中IL-4的浓度显著升高(P＜0.01),IFN-γ的浓度显著降低(P＜0.01),IFN-γ/IL-4比值显著降低(P＜0.01);疾病加证候(A)组和疾病(B)组大鼠均表现为肺组织中SP及其NK1受体mRNA表达显著增强(P＜0.01);疾病加证候(A)组和疾病(B)组大鼠均表现为肺、回肠组织中SP含量显著增加(P＜0.01),VIP含量显著减少(P＜0.01)。
2.与疾病(B)组比较,疾病加证候(A)组外周血白细胞计数、BALF中白细胞计数、嗜酸性粒细胞计数、血清IL-4浓度、BALF中IL-4浓度、肺组织SP及其NK1受体mRNA表达、肺组织及回肠组织SP含量均显著高于疾病(B)组(P＜0.05);BALF中IFN-γ浓度及IFN-γ/IL-4比值、肺组织及回肠组织VIP含量均显著低于疾病(B)组(P＜0.01或P＜0.05)。
3.与相应的疾病组比较,两中药(A1,B1)组血中及BALF中嗜酸性粒细胞计数显著降低(P＜0.01或P＜0.05)。其中,疾病加证候中药(A1)组BALF中嗜酸性粒细胞计数与对照组比较,已无显著差异(P＞0.05);疾病加证候中药(A1)组血中及BALF中白细胞计数显著降低(P＜0.01),疾病中药(B1)组BALF中白细胞计数显著降低(P＜0.01)。与相应的疾病组比较,两中药组(A1,B1)血中及BALF中IL-4的浓度显著降低(P＜0.01),IFN-γ的浓度显著升高(P＜0.01或P＜0.05),IFN-γ/IL-4比值显著升高(P＜0.01或P＜0.05)。其中,疾病加证候中药(A1)组血中IFN-γ浓度、BALF中IL-4的浓度及BALF中IFN-γ/IL-4比值与对照(C)组比较,已无显著差异(P＞0.05)。与相应的疾病组比较,两中药(A1,B1)组肺、回肠组织中SP含量显著减少(P＜0.01),VIP含量显著增加(P＜0.01),其中,疾病加证候中药(A1)组肺及回肠组织SP含量、疾病中药(B1)组回肠组织SP含量与对照(C)组比较,已无显著差异(P＞0.05)。与相应的疾病组比较,两中药(A1,B1)组肺组织中SP及其NK1受体mRNA表达显著减少(P＜0.01或P＜0.05)。其中,疾病加证候中药(A1)组肺组织SPmRNA表达水平、疾病中药(B1)组肺组织NK1受体mRNA表达水平与对照(C)组比较,已无显著差异(P＞0.05)。
4.各组大鼠血中IFN-γ/IL-4比值与肺组织SP含量间存在着明显的负相关(r=-0.698,P＜0.01);各组大鼠血中IFN-γ/IL-4比值与肺组织VIP含量间存在着明显的正相关(r=0.741,P＜0.01)。各组大鼠血中IFN-γ/IL-4比值与回肠组织SP含量间存在着明显的负相关(r=-0.627,P＜0.01);各组大鼠血中IFN-γ/IL-4比值与回肠组织VIP含量间存在着明显的正相关(r=0.766,P＜0.01)。各组大鼠BALF中IFN-γ/IL-4比值与肺组织SP含量间存在着明显的负相关(r=-0.688,P＜0.01);各组大鼠BALF中IFN-γ/IL-4比值与肺组织VIP含量间存在着明显的正相关(r=0.324,P＜0.01)。各组大鼠BALF中IFN-γ/IL-4比值与回肠组织SP含量间存在着明显的负相关(r=-0.615,P＜0.01);各组大鼠BALF中IFN-γ/IL-4比值与回肠组织VIP含量间存在着明显的正相关(r=0.679,P＜0.01)。各组大鼠肺组织中SP与VIP含量之间存在着明显的负相关(r=-0.678,P＜0.01);各组大鼠回肠组织中SP与VIP含量之间存在着明显的负相关(r=-0.620,P＜0.01)。各组大鼠肺组织与回肠组织中SP含量之间存在着明显的正相关(r=0.602,P＜0.01);各组大鼠肺组织与回肠组织中VIP含量之间存在着明显的正相关(r=0.608,P＜0.01)。
5.病理结果显示:疾病(B)组与疾病加证候(A)组均有不同程度的支气管及肺组织炎性病变,两中药(A1,B1)组的支气管及肺组织的炎性病变均有所减轻;疾病(B)组与疾病加证候(A)组均有不同程度的回肠组织炎性病变,两中药(A1,B1)组的回肠组织炎性病变程度有所减轻,但以疾病加证候治疗(A1)组的减轻程度明显。
结论:
1.哮喘及脾虚哮喘大鼠均存在着明显的气道局部或/和全身炎症性改变;哮喘及脾虚哮喘大鼠均存在着全身及气道局部免疫细胞因子IFN-γ、IL-4表达失衡;哮喘及脾虚哮喘大鼠均存在着肺、回肠组织内神经肽VIP、SP表达水平失衡;哮喘及脾虚哮喘大鼠均存在着肺组织内神经肽SP及其NK1受体mRNA表达水平失衡。
2.脾虚可以加重哮喘机体气道内炎性细胞聚集及外周血炎性细胞聚集,加重哮喘机体肺、回肠组织炎性病变,从而加重哮喘机体的局部及全身炎症反应;脾虚可以加重哮喘机体血清及BALF中免疫细胞因子IFN-γ、IL-4表达失衡,加重哮喘机体肺、回肠组织内神经肽VIP、SP表达水平失衡,加重肺组织内神经肽SP及其NK1受体mRNA表达水平失衡,从而加重哮喘机体的神经内分泌免疫调节失衡,成为哮喘反复发作的内在原因之一。
3.培土生金中药可以减轻模型大鼠气道局部及全身的炎性细胞浸润,下调血中及BALF中白细胞及嗜酸性粒细胞计数;培土生金中药可以调整全身及气道局部免疫细胞因子IFN-γ、IL-4表达失衡状态,上调血清及BALF中IFN-γ浓度,下调IL-4浓度,上调IFN-γ/IL-4比值;培土生金中药可以减轻肺、回肠组织内神经肽VIP、SP及NK1受体表达水平的失衡状态,下调肺组织中SP及其NK1受体mRNA表达水平,下调肺、回肠组织中SP含量表达水平,上调肺、回肠组织中VIP含量表达水平;培土生金中药能够减轻模型大鼠的大、小气道及肺、回肠组织的炎性病变。
4.肺、回肠组织神经肽VIP、SP表达异常及肺组织SP及其NK1受体mRNA表达的异常均与哮喘慢性气道炎症的形成与持续存在明显相关性,与气道局部及全身的炎症状态呈明显相关性,与气道局部及全身的免疫细胞因子IFN-γ、IL-4表达失衡状态亦呈明显相关性。这进一步证实了神经肽VIP、SP在哮喘发病过程中的重要作用,特别是介导哮喘的神经内分泌免疫调节失衡。肺组织与回肠组织的神经肽VIP、SP的表达水平存在着明显的相关性,从神经肽VIP、SP及神经内分泌免疫调节的角度进一步证实了肺脾二脏的密切关系。
OBJECTIVE:We attempted to investigate the imbalanced states of neuro-endocrine-immuno-modulation through study of the changes of neurikins and their receptors of the tissues of the lung and ileum in experimental rates with spleen-deficiency asthma and simple asthma;to investigate the influence of spleen deficiency on asthma by comparison of the changes of expressive levels between the two kinds of rat models,and thus to provide a new evidence for the view points of "the spleen deficiency as the key patho-mechanism".We attempted the effective mechanism of the therapy of "strengthening the earth and promoting the metal(SEPM)" in treatment of asthma through study of the influence of Chinese herbal remedy of SEPM to the expressive levels of neurikins and their receptors of the tissues of the lung and ileum in experimental rates with spleen-deficiency asthma and simple asthma;to investigate the modern connotation of SEPM theory and the close relationship between the lung and the spleen by studying the correlation between the expressive levels of neurikins and their receptors of the tissues of the lung and ileum in experimental rates with spleen-deficiency asthma and simple asthma.
MATERIAL AND METHODS,Taking the theory of traditional Chinese medicine (TCM) as the guide,we set up compound rat models with asthma of spleen deficiency by combination of establishing spleen deficiency animal models in TCM with establishing asthmatic animal models in Western medicine.The spleen models were set up by using the method of improper diet plus physical overstrain.The body weight and temperature of the anus of the animals were compared before and after the model establishment;and the symptoms of the animals were assessed by the standards for evaluation of spleen deficiency models in TCM.The successful animal models were determined by the statistical significance in the changes of their quantitative indexes and their symptomatologic accordance with the standards for evaluation of spleen deficiency.Asthmatic animal models were set up by using ovalbumin as the antigen,and sodium hydroxide and pertusis vaccine as the adjuvant.The asthmatic animals were set up by nebulization following the intraperitoneal injection of valbumin.The successful animal models were evaluated by the appearance of similar symptoms of human asthma in rats such as tachypnea,remarkable motion of nodding,strong construction of abdominal muscle,and occasional pulmonary rale and grunting.The successfully established animal models were given Chinese herbal remedy of SEPM for 12 days.Then the following determinations were conducted:white blood cell count(WBC) and eosinophil count(EOS) in peripheral blood and bronchial alveolar lavage fluid (BALF) by absolute counting;the contents of interleuki-4(IL-4) and interferon-γ(IFN-γ) in BALF by enzyme-linked immunoabsorbent assay(ELISA); the contents of substance P(SP) vasoactive intestinal peptide(VIP) by semi-quantitative analysis of immune-chemical assay;the expressions of SPmRNA and neurokinin 1(NK1) receptor mRNA by reverse transcription-polymerase chain reaction(RT-PCR).
RESULTS:
1.Compared with the control(C) group,the number of WBC and EOS of the peripheral blood and BALF in the compound(A) group and disease(B) group increased(P＜0.01);the content of IL-4 of BALF in group A and B decreased (P＜0.01),and the content of IFN-γof BALF decreased(P＜0.01),and the ratio of IFN-γ/IL-4 was remarkably decreased(P＜0.01).The mRNA expressions of SPmRNA and NK1 receptor mRNA in group A and B enhanced(P＜0.01);the content of SP in group A and B increased(P＜0.01) and the content of VIP obviously decreased(P＜0.01).
2.Compared with group B,the WBC in peripheral blood;the WBC and EOS in BALF;serum IL-4 level,IL-4 level in BALF,the expressions of SPmRNA and NK1 receptor mRNA,and the contents of SP of the lung and ileum tissues in group A all increased remarkably(P＜0.01 or P＜0.05).The level of IFN-γand ratio of IFN-γ/IL-4 in BALF,and the content of VIP in the lung and ileum tissues in group A all decreased remarkably(P＜0.05).
3.Compared with the correspondent disease groups,the EOS in blood and BALF in the two Chinese herbal groups(A1,B1) remarkably decreased(P＜0.01 or P＜0.05).And the EOS of BALF in group A1 had showed no significant difference from that in the control group(P＞0.05).The WBC and EOS of blood and BALF in group A1 obviously decreased(P＜0.01);the WBC of BALF in group B1 obviously decreased(P＜0.01).The levels of IL-4 of blood and BALF in both group A1 and B1 remarkably decreased(P＜0.01),but the levels of IFN-γobviously increased (P＜0.01 or P＜0.05);and the ratio of IFN-γ/IL-4 increased(P＜0.01 or P＜0.05).And the level of IFN-γof blood,the level of IL-4 of BALF,and the ratio of IFN-γ/IL-4 of BALF in group A1 had showed no significant difference from those in group C(P＞0.05).The expressions of SPmRNA and NK1 receptor mRNA in group A1 and B1 were remarkably decreased(P＜0.01 or P＜0.05).And the expressive levels of SPmRNA in group A1 and NK1 receptor mRNA in group B1 had showed no significant difference from those in group C(P＞0.05).The contents of SP in the lung and ileum in both group A1 and B1 obviously decreased(P＜0.01),but the content of VIP remarkably increased(P＜0.01).And the contents of SP in the lung and ileum in group A1,and the content of SP in the ileum in group B1 had showed no significant difference from those in group C(P＞0.05).
4.There was an obviously negative correlation between the ratio of IFN-γ/IL-4 of blood and the content of SP of the lung tissue in each group(r =-0.698,P＜0.01);there was an obviously positive correlation between the ratio of IFN-γ/IL-4 of blood and the content of VIP of the lung tissue(r=0.741, P＜0.01).There was an obviously negative correlation between the ratio of IFN-γ/IL-4 of blood and the content of SP of the ileum tissue in each group (r=-0.627,P＜0.01);there was an obviously positive correlation between the ratio of IFN-γ/IL-4 of blood and the content of VIP of the ileum tissue(r =0.766,P＜0.01).There was an obviously negative correlation between the ratio of IFN-γ/IL-4 of BALF and the content of SP of the lung tissue in each group (r=-0.688,P＜0.01);there was an obviously positive correlation between the ratio of IFN-γ/IL-4 of BALF and the content of VIP of the lung tissue(r= 0.324,P＜0.01).There was an obviously negative correlation between the ratio of IFN-γ/IL-4 of BALF and the content of SP of the ileum tissue in each group (r=-0.615,P＜0.01);there was an obviously positive correlation between the ratio of IFN-γ/IL-4 of BALF and the content of VIP of the ileum tissue(r= 0.679,P＜0.01).There was an obviously negative correlation between the contents of SP and VIP of the lung tissue in each group(r=-0.678,P＜0.01);there was an obviously positive correlation between the contents of SP and VIP of the lung tissue(r=-0.620,P＜0.01).There was an obviously negative correlation between the contents of SP and VIP of the ileum tissue in each group(r=0.602,P＜0.01);there was an obviously positive correlation between the contents of SP and VIP of the ileum tissue(r=0.608,P＜0.01).
5.The pathological results showed that there were bronchial and pulmonary inflammation at different degrees in both group B and A,and the inflammation in both group A1 and B1 had been reduced;there were ileal inflammation at different degrees in both group B and A,and the inflammation in both group A1 and B1 had been reduced,with the inflammation in group A1 was more obvious.
CONCLUSION:
1.There are,in asthmatic and spleen deficiency asthmatic rats,evident airway or/and general inflammation,expressive imbalance of general and airway immune cytokin IFN-γand IL-4,expressive imbalance of neurikins VIP and SP, and expressive imbalance of the SPmRNA and NK1mRNA receptors of the tissues of the lung and ileum,and the imbalance of neuro-endocrine-immuno-modulation.
2.Spleen deficiency can deteriorate the aggregation of inflammatory cells in the peripheral blood and airway of asthmatic rat models,and aggravate the pulmonary and ileal inflammation,and thus exacerbate the general and local inflammatory reaction of the asthmatic body.It can also exacerbate the imbalance of immune cytokins IFN-γand IL-4 in serum and BALF,and aggravate the expressive imbalance of neurikins VIP and SP and the SPmRNA and NK1mRNA receptors of the tissues of the lung and ileum,and thus make the imbalance of neuro-endocrine-immuno-modulation worse,being one of the inner causes for repeated attack of asthma.
3.Chinese remedy of SEPM can reduce the airway and general infiltration of inflammatory cells in rat models,down-regulate WBC and EOS in peripheral blood and BALF.It can also regulate the imbalanced state of general and local airway immune cytokins IFN-γand IL-4,up-regulare the content of IFN-γ, down-regulate the content of IL-4,and up-regulate the ratio of IFN-γ/IL-4. It can relieve the expressive imbalance of neurikins VIP,SP and NK1 receptors of the tissues of the lung and ileum,and down-regulate the expressive levels of SPmRNA and NK1mRNA receptors;down-regulate the expressive levels of SP and up-regulate the expressive levels of VIP in both the lung and ileum tissues. And the Chinese remedy of SEPM can reduce the tissue inflammation of the airways, lungs and ileum of the rat models.
4.The expressive imbalance of neurikins VIP and SP,and the SPmRNA and NK1mRNA receptors of the tissues of the lung and ileum is obviously correlated with the development and persistence of chronic airway inflammation in asthma, with general and local airway inflammatory state,and with the imbalanced state of general and local airway immune cytokins IFN-γand IL-4.This further reveals the important role of neurikins VIP and SP in development of asthma,especially in inducing imbalance of neuro-endocrine-immuno-modulation.There is evident correlation between the expressive levels of VIP and SP in the tissues of the lung and ileum,which further expounds the close relationship between the lung and the spleen from the angle of neurikins VIP and SP and neuro-endocrine-immuno-modulation.
引文
[1]Stock P,DeKruyffRH,Umetsu DT.Inhibition ofthe allergic response by regulatory T cells[J].CurrOpin Allergy Clin Immunol,2006,6(1):12-16
[2]Kim YR,Park Q,Yu BH.Changes in lymphocyte subsets after short-term pharmacotherapy in patients with panic disorder[J].Psychiatry Res,2004,128(2):183-190
[3]钟南山.哮喘临床防治研究的几点展望[J].中华医学信息导报,2004,15(24):2-3,8
[4]M auriC,Feldmann M,Williams RO,etal.Down-regulation of Th1-mediated pathology in experimentalarthritis by stimulations of Th2 arm ofthe immune response[J].Arthritis Rheum,2003,48(3):839-845
[5]江忠,田菊,杨志斌,等.玉屏风颗粒对哮喘患儿T淋巴细胞亚群的影响[J].中成药,2000,22(3):210-211
[6]万欢英,周敏,余莉,等.喘可治注射液治疗哮喘的疗效及其作用机制研究[J].中国新药与临床杂志,2005,24(3):210-212
[7]吴彬,俞建,王莹,等.补肾中药对哮喘缓解期患儿Th1/Th2平衡的影响[J].中国中西医结合杂志,2007,27(2):120-122
[8]于文涛,杨牧祥,王晓红,等.咳喘宁对支气管哮喘大鼠肺组织IFN-γ和IL-4含量的影响.中药药理与临床,2008,24(2):93-94
[9]张弘,王真,蔡宛如,等.清肺止哮汤对哮喘小鼠气道炎症的抑制及Th1/Th2平衡的影响[J].中华中医药学刊,2007,25(7):1475-1478
[10]赵杰,王竞.黄芪注射液对支气管哮喘模型大鼠的免疫调节作用[J].现代预防医学, 2007,34(15):2845-2846
[11]邢向晖,徐宁.平喘方对哮喘大鼠平喘作用的实验研究[J].中国中医药信息杂志,2002,9(12):17-19
[12]张庆祥,孙广仁,于少泓,等.温阳化饮方对哮喘病寒饮蕴肺证大鼠BALF中EOS的影响[J].中医药学刊,2006,24(8):1477-1478
[13]孙洪涛,林江涛,何雯雯.雷公藤和糖皮质激素对哮喘大鼠支气管肺泡灌洗液中嗜酸性粒细胞的影响[J].现代中西医结合杂志,2003,12(4):347-348,351
[14]旺建伟,李翼,徐国亭.苏子降气汤对哮喘大鼠核因子-κB表达及嗜酸性粒细胞数量的影响[J].中国实验方剂学杂志,2006,12(6):38-40
[15]高宝安,刘隆棣,江必武,等.芩荑合剂吸入对哮喘豚鼠气道嗜酸粒细胞凋亡的影响[J].中国中医基础医学杂志,2003,9(5):359-361
[16]伍参荣,刘贵云,李杰,等.治喘贴对实验豚鼠嗜酸粒细胞凋亡和对白细胞介素-5的影响[J].中华微生物和免疫学杂志,2004,24(7):549-551
[17]刘建秋,吴树亮,李竹英.喘康宁冲剂对支气管哮喘嗜酸粒细胞凋亡的实验研究[J].中医杂志,2003,20(1):56
[18]林永廉,林求诚.射干麻黄汤对实验性哮喘豚鼠嗜酸性粒细胞凋亡的影响[J].实用中医药杂志,2007,23(1):3-5
[19]胡作为,周燕萍,王鹏.止哮平喘方对哮喘豚鼠T淋巴细胞凋亡的作用[J].中国中医基础医学杂志,2004,10(1):31-33
[20]方向明,曹世宏.平喘合剂对哮喘豚鼠T淋巴细胞凋亡的影响[J].中国中医基础医学杂志,2003,9(3):184-186
[21]郭素华,罗风鸣,李灏来,等.加味肾气丸对Th1、Th2细胞凋亡及细胞因子表达影响的研究[J].贵阳中医学院学报,2004,26(1):48-50
[22]肖丽玲,伍参荣,唐小梅.定喘汤对哮喘大鼠NO、ET-1、IL-5的影响[J].中国实验方剂学杂志,2005,11(3):58-59
[23]何丽,郭盛,熊先敏,等.扶正化痰方对哮喘豚鼠肺组织NO、NOS浓度的影响[J].新中医,2003,35(3):78-79
[24]丁劲松,李继红,刘晓玲.知母宁对豚鼠哮喘的预防作用及其对内皮素和一氧化氮的影响[J].中国中医药科技,2007,14(2):89-90
[25]Faisy C,Naline E,Diehl JL,et al.The role of endothelin in the physiology and pathophysiology of airways[J].Rev Mal Respir,2002,19(3):341-355
[26]韩青,葛传生,郭梅.雷公藤内酯醇对哮喘豚鼠内皮素-1水平影响的动态观察[J].南京医科大学学报(自然科学版),2003,23(2):154-156
[27]刘恩顺,孙增涛.麻芩咳喘合剂治疗哮喘的临床疗效评价及机制研究[J].西部医学,2007,19(2):187-189
[28]王晓红,杨牧祥,于文涛,等.咳喘宁对支气管哮喘大鼠肺组织一氧化氮和内皮素-1含量的影响[J].中华中医药杂志,,22(1):12-15
[29]阳志晖,张建军,王林元,等.双龙胶囊对哮喘大鼠肺组织白三烯类化合物含量的影响[J].中国中西医结合杂志,2006,26(基础理论研究特集):43-46
[30]李学麟,陈少东,杨鸿.舒咳合剂治疗小儿咳嗽变异性哮喘与尿白三烯E4关系的研究[J].中医药学刊,2004,22(5):854-855
[31]朱慧华,陈燕萍,虞坚尔,等.辛苍汤对小儿支气管哮喘缓解期慢性气道炎症的治疗作用[J].中西医结合学报,2005,3(1):23-27
[32]姚迪,林建海,郏琴,等.白果对哮喘小鼠血清中白细胞介素4作用的研究[J].中国现代医学杂志,2006,14(12):5-7
[33]刘永刚,罗佳波.麻黄汤及拆方对哮喘小鼠5-脂质氧合酶激活蛋白、白介素4基因的表达和白三烯C4的影响[J].中国中药杂志,2007,32(3):246-249
[34]李明华,殷凯生,蔡映云主编.哮喘病学(第二版)[M],北京:人民卫生出版社.2005年6月:119
[35]童舜华,崔一丽,许建华.小青龙汤对哮喘大鼠肺组织β_2-肾上腺素能受体作用的研究[J].中国中医药科技,2001,8(2):77-79
[36]包照日格图,吴敦序,陈淑俊,等.补肾健脾中药对哮喘模型大鼠肺组织β-肾上腺素能受体的影响[J].中药新药与临床药理,2000,11(2):98-100
[37]刘军,鲍亚莉,秦雪冰.中药合剂对豚鼠哮喘模型气道及肺组织中神经肽的实验研究[J].中医药学报,2000,(1):71
[38]戴文军,赖新生.天灸疗法对支气管哮喘患者血浆SP和VIP含量的影响[J].针刺研究,2001,26(2):134-137
[39]沈虹,胡大文,唐纯志,等.穴位贴敷对支气管哮喘患者血浆SP和VIP含量的影响[J].河南中医,2002,22(2):20-21
[40]汪珊,梁仁.中药地龙药理与哮喘气道重建相关性研究[J].广东药学院学报,2004, 20(1):60-62
[41]林琳,要全保,于素霞,等.止喘胶囊对实验性哮喘动物模型气道重建的影响[J].中国中西医结合杂志,2001,2(8):599-601
[42]李志奎,王长征,钱桂生.雷公藤甲素对哮喘豚鼠嗜酸性粒细胞凋亡及Fas,Bcl-2mRNA表达的影响[J].中国新药与临床杂志,2002,21(5):261-264
[43]范欣生,许冬青,王明艳,等.复方辛夷平喘液对哮喘豚鼠肺内LCA及淋巴细胞fas表达的影响[J].江苏中医药,2005,26(4):50-52
[44]贯剑,郑湘瑞,何裕民,等.扶正固本法对青少年哮喘患者基因表达的影响[J].复旦学报(医学版),2006,33(6):834-836
[45]赵福东,董竞成,崔焱,等.淫羊藿对哮喘大鼠神经内分泌免疫网络若干指标的影响[J].中国实验方剂学杂志,2007,13(9):44-46
[46]董竞成,赵福东,谢瑾玉,等.黄芪对哮喘大鼠神经内分泌免疫网络相关指标的影响[J].中国中西医结合杂志,2007,27(7):619-622
[47]许建华,范忠泽,吴敦序,等.补肾定喘汤对哮喘大鼠肺组织糖皮质激素受体及血浆皮质酮、ACTH的影响[J].中国中医基础医学杂志,2003,9(1):27-29
[48]毛昌淳,方肇勤,吴敦序,等.补肾定喘汤对哮喘大鼠血CORT、下丘脑CRHmRNA表达的影响[J].中成药,2003,25(6):468-470
[49]宋曦,张有花,周兆山.中药治疗支气管哮喘作用机理实验研究进展[J].中国中医药信息杂志,2006,13(1):104-106
[1]李德新,王晓明,易杰,等.脾虚证对生物膜结构与功能影响的实验研究[J].辽宁中医杂志,1993,20(6):39-43
[2]张焱.培土生金法对脾虚哮喘大鼠一氧化氮水平改变影响的实验研究(硕士学位论文).辽宁:辽宁中医学院,2002
[3]吕国平,崔德健,郭英江,等.介绍一种建立大鼠哮喘模型的实验方法[J].中华结核和呼吸杂志,1995,18(6):377-378
[4]Reed KL,Fruin A B,Bishop-Bartolomei kk,et al.Neurokinin-1 Receptor and Substance P Messenger RNA Levels Increase during Intraabdominal Adhesion Formation[J].J Surg Res,2002;108(1):165-168
[5]杨保林,武维屏,王硕仁,等.心理应激对哮喘大鼠肺内神经肽及NK1受体mRNA表达的影响[J].中国中医基础医学杂志,2004,10(4):15-17
[6]骆仙芳,蔡宛如.王会仍辨治支气管哮喘的经验[J].浙江中医杂志,1999,(4):147-148
[7]熊耀康,俞冰.哮喘临床研究进展[J].浙江中医学院学报,2001,25(3):5-7
[8]傅志红.洪广祥教授治肺系疾病探要[J].新中医,1999,31(1):11-13
[9]张爱国.从痰瘀论治支气管哮喘[J].山东中医杂志,2002,21(10):632-633
[10]朱慧华,虞坚尔.从痰瘀论治儿童支气管哮喘机理初探[J].上海中医药杂志,2006,40(11):40-41
[11]刘玉山,张爱华,王玉民,等.射干麻黄汤加味治疗小儿哮喘103例疗效观察[J].河北中医,2001,23(6):454
[12]杨定保.小儿哮喘辨治三法[J].湖北中医杂志,2001,23(4):21-22
[15]刘锁超.支气管哮喘临证心得[J].新中医,2002,34(8):68
[16]张洪春,晁恩祥.疏风解痉法治疗过敏性支气管哮喘的临床研究[J].中国中医急症,1998,7(2):54-58
[17]陈可静,郝小梅,刘梅.治风化痰平哮方治疗小儿支气管哮喘发作期临床研究[J].中国中西医结合杂志,2003,23(9):673-675
[13]陈碧霞.清肺化痰汤治疗小儿热性哮喘42例[J].四川中医,2002,20(5):56-57
[14]许光辉,罗彬.泻肺平喘法配合西药治疗支气管哮喘40例[J].陕西中医,2005,26(8):744-745
[18]崔红生,牛常霞.武维屏辨治支气管哮喘经验举隅[J].北京中医药大学学报,2001,24(6):66-67
[19]郭军雄,呼兴华.从肝论治小儿支气管哮喘[J].中医儿科杂志,2006,2(6):22-23,30
[20]续海卿.从肝胃论治哮喘的体会[J].四川中医,2003,21(5):22
[21]陈宁,王清海,黄琳,等.胃不和与支气管哮喘的关系初探[J].广东医学,2001,22(8):755-756
[22]易向明,哮喘发作伴大便异常的观察及临床意义[J].四川中医,1995,(3):7-8
[23]顾维明.通腑法治疗急症临床体会[J].陕西中医,2001,22(11):703-704
[24]韩镭,于金霞.赵子贤治疗哮喘急性发作用药特色浅析[J].浙江中医杂志,2001,(7):282-283
[25]哈木拉提·吾甫尔,李风森,艾买江·吐尼亚孜.“肺与大肠相表里”的科学性及其对哮喘治疗的指导意义[J].新疆医科大学学报,2000,23(2):151-152
[26]夏永良,王卉,李德新.论“脾统四脏”[J].中国中医基础医学杂志,2001,7(10):69-71
[27]周静.哮喘的脏腑论治[J].四川中医,2003,21(3):9-10
[28]夏永良,王彩霞.哮喘缓解期从脾论治机理的研究[J].中医药学刊,2003,21(7):1194,1196,1199
[29]曹宏,张桂菊.小儿哮喘从痰辨治体会[J].四川中医,2005,23(4):67-68
[30]吴敏.700例哮喘儿童患病情况及中医辨证资料计算机数据库统计分析[J].上海中医药杂志,1992(6):25
[31]王明明,汪受传.玉屏风散预防小儿哮喘临床效果及机理探讨[J].时珍国医国药,2001,12(5):457-458
[32]符晓,王大海,龚兴宏,等.补肾温肺胶囊治疗支气管哮喘缓解期患儿的疗效观察[J].中国中西医结合杂志,2001,21(9):699-700
[33]倪伟,于素霞,王宏长,等.缓解期哮喘患者肾阳虚与气道炎症关系的研究-附175例病例分析[J].中国中医基础医学杂志,2001,7(6):60-61
[34]许得盛,沈自尹,胡国让,等.温阳片防治支气管哮喘远期疗效观察[J].上海中医药杂志,1989,(12):4-5
[35]张子明.朱佳运用补肾法治疗哮喘的经验[J].辽宁中医杂志,2007,34(2):132-133
[36]李明华,殷凯生,蔡映云主编.哮喘病学(第二版)[M],2005:62,111-113,118,121,594
[37]Wang HY,Shen HH,Lee J J,et al.CD69 expression on airway eos inophils and airway inflammation in a murine model[J].Chin Med J (Engl),2006;119(23):1983-1990
[38]Kumar RK,Herbert C,Webb DC,et al.Effects of anticytokine therapy in a mouse model of chronic asthma[J].Am J Respir Crit Care Med,2005;172(2):256
[39]Fos ter PS,Webb DC,Yang M,et al.Dis sociation of T helper type 2cytokine-dependent airway les ions from signal transducer and activator of transcription 6 signalling in experimental chronic asthma[J].Clin Exp Allergy,2003;33(5):688-695
[40]Cockcroft DW.Mechanisms of airway hyperrespons iveness Davis BE[J].J Allergy Clin Immunol,2007;119(1):254-255
[41]李凝.神经肽和哮喘[J].国外医学呼吸系统分册,2002,22(1):45-48
[42]谢启文主编.现代神经内分泌学[M].上海:上海医科大学出版社,1999:60
[43]Stimson WH.Oestrogenandhuman Tlym Phoeytes:Preseneeofs Peeifiereee Ptors in the T-suP Pressor/eytotoxie subset[J].Seand J Immunol,1998:28:345-350
[44]宫兆华.哮喘神经内分泌免度网络紊乱及补肾益气药对其调节作用的研究(博士学位论文).上海:复旦大学,2005
[45]李明华.哮喘病的新概念和治疗研究进展[J].中国临床医生,2004,32(2):2-4
[46]Heaney LG,Robinson DS.Severe asthma treatment:need for characterizing patients[J].Lancet,2005,365(9463):974-976
[47]Bloom JW,Chaeho J,Lohman IC,et al.Diferential control of essinophil survival by glucocorticoids[J].Apoptosis,2004,9(1):97-104
[48]Scanlon PD,Connett JE,Wise RA,et al.Loss of bone density with inhaled triamcinolone in Lung Health Study Ⅱ[J].Am J Respir Crit Care Med,2004;170:1302-1309
[49]O' Connor BJ,Lofdahl CG,Balter M,et al.Zileuton added to low-dose inhaled beclomethasone for the treatment of moderate to severe persistent asthma[J]. Respir Med,2007;101:1088-1096
[50]刘自力.培土生金法对脾虚哮喘大鼠嗜酸细胞凋亡及对FasmRNA、Bcl-2mRNA表达的影响(博士学位论文).辽宁:辽宁中医药大学,2006
[51]苗明三,主编.实验动物和动物实验技术(第一版)[M].北京:中国中医药出版社,1997:100
[52]苗会,薛全福,庄逢源,等.哮喘大鼠动物模型的制备[J].基础医学与临床,1998,18(1):72-78
[53]Joseph-Bowen J,de Klerk N,Holt PG,et al.Relationship of asthma,atopy,and bronchial responsivenese to serum eosinophil cationic proteins in early childhood[J].J Allergy Clin Immunol,2004;114:1040-1042
[54]Fabbri LM,Romagnoli M,Corbetta L,et al.Differences in airway inflammation in patients with fixed airflow obstruction due to asthma or chronic obstructive pulmonary disease[J].Am J Respir Crit Care Med,2003;167:418-424
[55]Peng T,Hac L,Madri JA,et al.Role of C5 in the development of airway inflammation,airway hyperresponsiveness,and ongoing airway response[J].J Clin Invest,2005;115:1590-1595
[56]Berro AI,Perry GA,Agrawal DK.Increased expression and activation of CD30induce apoptosis in human blood cosinophils[J].J Immunol,2004;173:2174-2178
[57]郭淑文,孟庆刚,王硕仁.病证结合模型建立的思路[J].中医药学报,2001,29(1):2-4
[58]Wood N,WhittersMJ,Jacobson BA,et al.Enhanced interleukin(IL)-13responses in mice lacking IL-13 recep tor alpha 2[J].J Exp Med,2003;197(6):703-709
[59]LI Ya-fei,GUO Bo-tao,HAN Jia-xin,et al.Association between interleukin-4polymorphisms and asthma[J].Journal of Medical Colleges of PLA,2007;22(5):312-319
[60]Ngoc PL,Gold DR,TzianabesAO,et al.Cytokines,allergy,and asthma[J].Curt Op in Allergy Clin Immunol,2005;5(2):161-166
[61]Romagnani S.Cytokines and chemoattractants in allergic inflammation[J].Mol Immunol,2002;38(12-13):881-885
[62]王文建,杨莉.地塞米松对支气管哮喘模型大鼠气道重建的影响[J].郑州大学学报:医学版,2007,42(1):130-133
[63]SugitaM,Kuribayashi K,Nakagomi T,et al.Allergic bronchial asthma:airway inflammation and hyperresponsiveness[J].Intern Med,2003;42(8):636-643
[64]Shen H,O' Byrne PM,Ellis R,et al.The effects of intranasalbudesonide on allergen-induced production of interleukin-5 and eotaxin,airways,blood,and bone marrow eosinophilia,andeosinophil progenitor expansion in sensitized mice[J].Am J Resp ir Crit CareMed,2002;166(2):146-153
[65]Joos GF,PR Germonpre,JC Kips,et al.Sensory neuropeptides and the human lower airways:present state and future directions[J].Eur Respir J,1994;7:1161-1171
[66]Barnes PJ.Overview of neural mechanisms in asthma[J].Pulm Pharmacol,1995;8(4/5):151-159
[67]Spina D.Airway sensory nerves:a burning issue in asthma?[J].Thorax,1996;51(3):335-337
[68]高振,阿地力江·阿布力米提,哈木拉提·吾甫尔.哮喘的下丘脑-垂体-肾上腺轴紊乱及中医药介入的影响[J].实用医学杂志,2008,24(11):1855-1856
[69]谢启文主编.神经肽(第一版)[M].上海:复旦大学出版社.2004:164,324,331-333,344
[70]Maghni K,Michoud MC,Alles M,et al.Airway smooth muscle cells express functional neurokinin-1 receptors and the nerve-derived preprotachykinin-a gene:regulation by passive sensitization[J].Am J Respir Cell Mol Biol,2003;28(1):103-110
[71]Groneberg DA,Quarcoo D,Frossard N,et al.Neurogenic mechanisms in bronchial inflammatory diseases[J].Allergy,2004;59:1139-1152
[72]Joos GF,Germonpre PR,Kips JC,et al.Sensory neuropeptides and the human lower airways:present state and future directions[J].Eur Respir J,1994;7:1161-1171
[73]Robinson DS.The role of the mast cell in asthma:induction of airway hyperresponsiveness by interaction with smooth muscle[J]? J Allergy Clin Immunol,2004;114:58-65
[74]Opconner TM,Opconnell J,O' brien D I et al.The Role of substance P in inflammatory disease[J].J Cell Physiol,2004;201:167-180
[75]De Swert KO,Toumoy KG,Joos GF,et al.The role of the tachykinin NK1 receptor in airway changes in a mouse model of allergic asthma[J].J Allergy Clin Immunol,2004;113(6):1093-1097
[76]Groneberg DA,Quarcoo D,Frossard N et al.Neurogenic mechanisms in bronchial inflammatory diseases[J].Allergy,2004;59:1139-1152
[77]van der Kleij H P,Kraneveld A D,Redegeld F A,et al.The tachykinin NK1receptor is crucial for the development of nonatopic airway inflammation and hyperresponsiveness[J].Eur J Pharmacol,2003;476(3):249-255
[78]李淼.RT-PCR检测NK1mRNA在哮喘豚鼠气道表达的研究[J].辽宁医学杂志,2008,22(4):191-192
[79]李利生,曲瑞瑶,郭华,等.冷应激大鼠血浆及回肠P物质和VIP的变化[J].首都医科大学学报,2002,23(2):113-114
[80]张忠兵,张学庸,陈希陶,等.脾虚泄泻患者肠粘膜中SP和VIP初步探讨[J].中西医结合杂志,1991,11(3):144-146,132
[81]刘素梅,曲瑞瑶,王伟,等.实验性脾虚证大鼠胃窦部SP、VIP、CGRP含量的研究[J].首都医科大学学报,1998,19(2):105-108
[82]曲瑞瑶,刘素梅,王伟,等.实验性脾虚证大鼠胃肠动力学与胃肠肽关系的研究[J].中国中西医结合杂志,1998,18(基础理论研究特集):237-240,391
[83]刘素梅,曲瑞瑶,曾文红,等.实验性脾虚证大鼠小肠P物质、血管活性肠肽、降钙素基因相关肽含量的变化[J].中国中西医结合杂志,1997,17(基础理论研究特集):265-267,308
[84]郭华,曲瑞瑶,常延滨,等.实验性脾虚证大鼠回肠P物质和血管活性肠肽含量的变化[J].首都医科大学学报,2001,22(3):199-201
[85]李利生,曲瑞瑶,王伟,等.实验性脾虚大鼠与冷应激大鼠回肠段SP、VIP含量的变化[J].深圳中西医结合杂志,2002,12(2):75-76,79
[86]Garcia-V illar-R;Dupuis-C;M artino lle-Jpect.Functional evidence fo r NO-synthase activationby substance P th rough a mechanism no t invo lying classical tachyk in in receptors in guinea-pig ileum in vitro[J].BR-J-Pharmacal.1996;118(5):1253-1256
[87]Cayabyab-FS,J imenez-M,V ergera-Petal.Influence of nitric oxide and vasoactive intestinal pep tide on the spontaneous and triggered electrical and mechanical activities of the canine ileum[J].Can-J-Physiol-Pharmacol.1997;75(5):383-387
[88]周吕主编.胃肠生理学[M].北京:科学出版社,1991:738
[89]陈华,盛丽先.胃肠激素与脾虚型小儿厌食症关系探讨[J].中国中西医结合脾胃杂志,2000,8(3):153-154
[90]高碧珍,黄爱民,李灿东,等.不同证候胃炎模型血生化指标的实验研究[J].福建中医学院学报,2005,15(1):30-33
[91]吴娟,田德禄.慢性浅表性胃炎脾胃湿热证胃黏膜G、D细胞变化及胃泌素、生长抑素的表达[J].世界华人消化杂志,2008,16(34):3840-3843
[92]钱会南,吴海霞,王乐,等.健脾与补肾对脑内神经肽Y水平和基因表达的影响[J].中国中医药信息杂志,2006,13(4):29-31
[93]刘自力,张庆荣,吴兆利.论脾虚为哮喘宿根[J].中医药学刊,2006,24(2):294-295
[94]张庆荣,谷峰,赵世芬.脾虚对哮喘氧化抗氧化平衡改变的影响[J].辽宁中医杂志,1999,26(12):574-575
[95]孙晓玲,闫英海,张庆荣.培土生金对脾虚哮喘大鼠气道环核苷酸水平的影响[J].辽宁中医杂志,2001,28(8):507-508
[96]张庆荣,陈金,赵世芬.脾虚对哮喘大鼠嗜酸细胞凋亡的影响[J].中医药学刊,2005,23(12):2129-2130
[97]张焱,张庆荣.培土生金法对脾虚哮喘大鼠一氧化氮水平改变影响的实验研究[J].吉林中医药,2007,27(10):52-54
[98]Groneberg DA,Quarcoo D,Frossard N,et al.Neurogenic mechanisms in bronchial inflammatory diseases[J].Allergy,2004;59:1139-1141
[99]Joos GF,Germonpre PR,Pauwels RA.Neurogenic inflammation in human airways:is it important?[J].Thorax,1995;50(3):217-219
[100]俞茹云,王彤,殷凯生,等.哮喘大鼠肺内感觉神经-内分泌细胞分布和密度的变化[J].南京医科大学学报(自然科学版),2008,28(30):316-320
[101]张少军,童瑶.肺与大肠相表里的现代研究及评析[J].中医函授通讯,2000,19(3):14-16
[102]E M stermbetg.Neuroendoerine regulation of autoimmune/inflammatory disease[J].Journal of Endocrinology,2001;169:29-33
[103]陈晓萍,徐远扬.神经、内分泌、免疫网络的通用生物学语言[J].自然杂志,2002,24(4):194
[104]刘永琦,王文.虚证的免疫学本质[J].中国中医基础医学杂志,2003,9(5):7-10,13
[105]刘文俊,李荣兴.中医五脏实质是人体生命本质[J].中国中医基础医学杂志,1995,1(3):16
[2]Kim YR,Park Q,Yu BH.Changes in lymphocyte subsets after short-term pharmacotherapy in patients with panic disorder[J].Psychiatry Res,2004,128(2):183-190
[3]钟南山.哮喘临床防治研究的几点展望[J].中华医学信息导报,2004,15(24):2-3,8
[4]M auriC,Feldmann M,Williams RO,etal.Down-regulation of Th1-mediated pathology in experimentalarthritis by stimulations of Th2 arm ofthe immune response[J].Arthritis Rheum,2003,48(3):839-845
[5]江忠,田菊,杨志斌,等.玉屏风颗粒对哮喘患儿T淋巴细胞亚群的影响[J].中成药,2000,22(3):210-211
[6]万欢英,周敏,余莉,等.喘可治注射液治疗哮喘的疗效及其作用机制研究[J].中国新药与临床杂志,2005,24(3):210-212
[7]吴彬,俞建,王莹,等.补肾中药对哮喘缓解期患儿Th1/Th2平衡的影响[J].中国中西医结合杂志,2007,27(2):120-122
[8]于文涛,杨牧祥,王晓红,等.咳喘宁对支气管哮喘大鼠肺组织IFN-γ和IL-4含量的影响.中药药理与临床,2008,24(2):93-94
[9]张弘,王真,蔡宛如,等.清肺止哮汤对哮喘小鼠气道炎症的抑制及Th1/Th2平衡的影响[J].中华中医药学刊,2007,25(7):1475-1478
[10]赵杰,王竞.黄芪注射液对支气管哮喘模型大鼠的免疫调节作用[J].现代预防医学, 2007,34(15):2845-2846
[11]邢向晖,徐宁.平喘方对哮喘大鼠平喘作用的实验研究[J].中国中医药信息杂志,2002,9(12):17-19
[12]张庆祥,孙广仁,于少泓,等.温阳化饮方对哮喘病寒饮蕴肺证大鼠BALF中EOS的影响[J].中医药学刊,2006,24(8):1477-1478
[13]孙洪涛,林江涛,何雯雯.雷公藤和糖皮质激素对哮喘大鼠支气管肺泡灌洗液中嗜酸性粒细胞的影响[J].现代中西医结合杂志,2003,12(4):347-348,351
[14]旺建伟,李翼,徐国亭.苏子降气汤对哮喘大鼠核因子-κB表达及嗜酸性粒细胞数量的影响[J].中国实验方剂学杂志,2006,12(6):38-40
[15]高宝安,刘隆棣,江必武,等.芩荑合剂吸入对哮喘豚鼠气道嗜酸粒细胞凋亡的影响[J].中国中医基础医学杂志,2003,9(5):359-361
[16]伍参荣,刘贵云,李杰,等.治喘贴对实验豚鼠嗜酸粒细胞凋亡和对白细胞介素-5的影响[J].中华微生物和免疫学杂志,2004,24(7):549-551
[17]刘建秋,吴树亮,李竹英.喘康宁冲剂对支气管哮喘嗜酸粒细胞凋亡的实验研究[J].中医杂志,2003,20(1):56
[18]林永廉,林求诚.射干麻黄汤对实验性哮喘豚鼠嗜酸性粒细胞凋亡的影响[J].实用中医药杂志,2007,23(1):3-5
[19]胡作为,周燕萍,王鹏.止哮平喘方对哮喘豚鼠T淋巴细胞凋亡的作用[J].中国中医基础医学杂志,2004,10(1):31-33
[20]方向明,曹世宏.平喘合剂对哮喘豚鼠T淋巴细胞凋亡的影响[J].中国中医基础医学杂志,2003,9(3):184-186
[21]郭素华,罗风鸣,李灏来,等.加味肾气丸对Th1、Th2细胞凋亡及细胞因子表达影响的研究[J].贵阳中医学院学报,2004,26(1):48-50
[22]肖丽玲,伍参荣,唐小梅.定喘汤对哮喘大鼠NO、ET-1、IL-5的影响[J].中国实验方剂学杂志,2005,11(3):58-59
[23]何丽,郭盛,熊先敏,等.扶正化痰方对哮喘豚鼠肺组织NO、NOS浓度的影响[J].新中医,2003,35(3):78-79
[24]丁劲松,李继红,刘晓玲.知母宁对豚鼠哮喘的预防作用及其对内皮素和一氧化氮的影响[J].中国中医药科技,2007,14(2):89-90
[25]Faisy C,Naline E,Diehl JL,et al.The role of endothelin in the physiology and pathophysiology of airways[J].Rev Mal Respir,2002,19(3):341-355
[26]韩青,葛传生,郭梅.雷公藤内酯醇对哮喘豚鼠内皮素-1水平影响的动态观察[J].南京医科大学学报(自然科学版),2003,23(2):154-156
[27]刘恩顺,孙增涛.麻芩咳喘合剂治疗哮喘的临床疗效评价及机制研究[J].西部医学,2007,19(2):187-189
[28]王晓红,杨牧祥,于文涛,等.咳喘宁对支气管哮喘大鼠肺组织一氧化氮和内皮素-1含量的影响[J].中华中医药杂志,,22(1):12-15
[29]阳志晖,张建军,王林元,等.双龙胶囊对哮喘大鼠肺组织白三烯类化合物含量的影响[J].中国中西医结合杂志,2006,26(基础理论研究特集):43-46
[30]李学麟,陈少东,杨鸿.舒咳合剂治疗小儿咳嗽变异性哮喘与尿白三烯E4关系的研究[J].中医药学刊,2004,22(5):854-855
[31]朱慧华,陈燕萍,虞坚尔,等.辛苍汤对小儿支气管哮喘缓解期慢性气道炎症的治疗作用[J].中西医结合学报,2005,3(1):23-27
[32]姚迪,林建海,郏琴,等.白果对哮喘小鼠血清中白细胞介素4作用的研究[J].中国现代医学杂志,2006,14(12):5-7
[33]刘永刚,罗佳波.麻黄汤及拆方对哮喘小鼠5-脂质氧合酶激活蛋白、白介素4基因的表达和白三烯C4的影响[J].中国中药杂志,2007,32(3):246-249
[34]李明华,殷凯生,蔡映云主编.哮喘病学(第二版)[M],北京:人民卫生出版社.2005年6月:119
[35]童舜华,崔一丽,许建华.小青龙汤对哮喘大鼠肺组织β_2-肾上腺素能受体作用的研究[J].中国中医药科技,2001,8(2):77-79
[36]包照日格图,吴敦序,陈淑俊,等.补肾健脾中药对哮喘模型大鼠肺组织β-肾上腺素能受体的影响[J].中药新药与临床药理,2000,11(2):98-100
[37]刘军,鲍亚莉,秦雪冰.中药合剂对豚鼠哮喘模型气道及肺组织中神经肽的实验研究[J].中医药学报,2000,(1):71
[38]戴文军,赖新生.天灸疗法对支气管哮喘患者血浆SP和VIP含量的影响[J].针刺研究,2001,26(2):134-137
[39]沈虹,胡大文,唐纯志,等.穴位贴敷对支气管哮喘患者血浆SP和VIP含量的影响[J].河南中医,2002,22(2):20-21
[40]汪珊,梁仁.中药地龙药理与哮喘气道重建相关性研究[J].广东药学院学报,2004, 20(1):60-62
[41]林琳,要全保,于素霞,等.止喘胶囊对实验性哮喘动物模型气道重建的影响[J].中国中西医结合杂志,2001,2(8):599-601
[42]李志奎,王长征,钱桂生.雷公藤甲素对哮喘豚鼠嗜酸性粒细胞凋亡及Fas,Bcl-2mRNA表达的影响[J].中国新药与临床杂志,2002,21(5):261-264
[43]范欣生,许冬青,王明艳,等.复方辛夷平喘液对哮喘豚鼠肺内LCA及淋巴细胞fas表达的影响[J].江苏中医药,2005,26(4):50-52
[44]贯剑,郑湘瑞,何裕民,等.扶正固本法对青少年哮喘患者基因表达的影响[J].复旦学报(医学版),2006,33(6):834-836
[45]赵福东,董竞成,崔焱,等.淫羊藿对哮喘大鼠神经内分泌免疫网络若干指标的影响[J].中国实验方剂学杂志,2007,13(9):44-46
[46]董竞成,赵福东,谢瑾玉,等.黄芪对哮喘大鼠神经内分泌免疫网络相关指标的影响[J].中国中西医结合杂志,2007,27(7):619-622
[47]许建华,范忠泽,吴敦序,等.补肾定喘汤对哮喘大鼠肺组织糖皮质激素受体及血浆皮质酮、ACTH的影响[J].中国中医基础医学杂志,2003,9(1):27-29
[48]毛昌淳,方肇勤,吴敦序,等.补肾定喘汤对哮喘大鼠血CORT、下丘脑CRHmRNA表达的影响[J].中成药,2003,25(6):468-470
[49]宋曦,张有花,周兆山.中药治疗支气管哮喘作用机理实验研究进展[J].中国中医药信息杂志,2006,13(1):104-106
[1]李德新,王晓明,易杰,等.脾虚证对生物膜结构与功能影响的实验研究[J].辽宁中医杂志,1993,20(6):39-43
[2]张焱.培土生金法对脾虚哮喘大鼠一氧化氮水平改变影响的实验研究(硕士学位论文).辽宁:辽宁中医学院,2002
[3]吕国平,崔德健,郭英江,等.介绍一种建立大鼠哮喘模型的实验方法[J].中华结核和呼吸杂志,1995,18(6):377-378
[4]Reed KL,Fruin A B,Bishop-Bartolomei kk,et al.Neurokinin-1 Receptor and Substance P Messenger RNA Levels Increase during Intraabdominal Adhesion Formation[J].J Surg Res,2002;108(1):165-168
[5]杨保林,武维屏,王硕仁,等.心理应激对哮喘大鼠肺内神经肽及NK1受体mRNA表达的影响[J].中国中医基础医学杂志,2004,10(4):15-17
[6]骆仙芳,蔡宛如.王会仍辨治支气管哮喘的经验[J].浙江中医杂志,1999,(4):147-148
[7]熊耀康,俞冰.哮喘临床研究进展[J].浙江中医学院学报,2001,25(3):5-7
[8]傅志红.洪广祥教授治肺系疾病探要[J].新中医,1999,31(1):11-13
[9]张爱国.从痰瘀论治支气管哮喘[J].山东中医杂志,2002,21(10):632-633
[10]朱慧华,虞坚尔.从痰瘀论治儿童支气管哮喘机理初探[J].上海中医药杂志,2006,40(11):40-41
[11]刘玉山,张爱华,王玉民,等.射干麻黄汤加味治疗小儿哮喘103例疗效观察[J].河北中医,2001,23(6):454
[12]杨定保.小儿哮喘辨治三法[J].湖北中医杂志,2001,23(4):21-22
[15]刘锁超.支气管哮喘临证心得[J].新中医,2002,34(8):68
[16]张洪春,晁恩祥.疏风解痉法治疗过敏性支气管哮喘的临床研究[J].中国中医急症,1998,7(2):54-58
[17]陈可静,郝小梅,刘梅.治风化痰平哮方治疗小儿支气管哮喘发作期临床研究[J].中国中西医结合杂志,2003,23(9):673-675
[13]陈碧霞.清肺化痰汤治疗小儿热性哮喘42例[J].四川中医,2002,20(5):56-57
[14]许光辉,罗彬.泻肺平喘法配合西药治疗支气管哮喘40例[J].陕西中医,2005,26(8):744-745
[18]崔红生,牛常霞.武维屏辨治支气管哮喘经验举隅[J].北京中医药大学学报,2001,24(6):66-67
[19]郭军雄,呼兴华.从肝论治小儿支气管哮喘[J].中医儿科杂志,2006,2(6):22-23,30
[20]续海卿.从肝胃论治哮喘的体会[J].四川中医,2003,21(5):22
[21]陈宁,王清海,黄琳,等.胃不和与支气管哮喘的关系初探[J].广东医学,2001,22(8):755-756
[22]易向明,哮喘发作伴大便异常的观察及临床意义[J].四川中医,1995,(3):7-8
[23]顾维明.通腑法治疗急症临床体会[J].陕西中医,2001,22(11):703-704
[24]韩镭,于金霞.赵子贤治疗哮喘急性发作用药特色浅析[J].浙江中医杂志,2001,(7):282-283
[25]哈木拉提·吾甫尔,李风森,艾买江·吐尼亚孜.“肺与大肠相表里”的科学性及其对哮喘治疗的指导意义[J].新疆医科大学学报,2000,23(2):151-152
[26]夏永良,王卉,李德新.论“脾统四脏”[J].中国中医基础医学杂志,2001,7(10):69-71
[27]周静.哮喘的脏腑论治[J].四川中医,2003,21(3):9-10
[28]夏永良,王彩霞.哮喘缓解期从脾论治机理的研究[J].中医药学刊,2003,21(7):1194,1196,1199
[29]曹宏,张桂菊.小儿哮喘从痰辨治体会[J].四川中医,2005,23(4):67-68
[30]吴敏.700例哮喘儿童患病情况及中医辨证资料计算机数据库统计分析[J].上海中医药杂志,1992(6):25
[31]王明明,汪受传.玉屏风散预防小儿哮喘临床效果及机理探讨[J].时珍国医国药,2001,12(5):457-458
[32]符晓,王大海,龚兴宏,等.补肾温肺胶囊治疗支气管哮喘缓解期患儿的疗效观察[J].中国中西医结合杂志,2001,21(9):699-700
[33]倪伟,于素霞,王宏长,等.缓解期哮喘患者肾阳虚与气道炎症关系的研究-附175例病例分析[J].中国中医基础医学杂志,2001,7(6):60-61
[34]许得盛,沈自尹,胡国让,等.温阳片防治支气管哮喘远期疗效观察[J].上海中医药杂志,1989,(12):4-5
[35]张子明.朱佳运用补肾法治疗哮喘的经验[J].辽宁中医杂志,2007,34(2):132-133
[36]李明华,殷凯生,蔡映云主编.哮喘病学(第二版)[M],2005:62,111-113,118,121,594
[37]Wang HY,Shen HH,Lee J J,et al.CD69 expression on airway eos inophils and airway inflammation in a murine model[J].Chin Med J (Engl),2006;119(23):1983-1990
[38]Kumar RK,Herbert C,Webb DC,et al.Effects of anticytokine therapy in a mouse model of chronic asthma[J].Am J Respir Crit Care Med,2005;172(2):256
[39]Fos ter PS,Webb DC,Yang M,et al.Dis sociation of T helper type 2cytokine-dependent airway les ions from signal transducer and activator of transcription 6 signalling in experimental chronic asthma[J].Clin Exp Allergy,2003;33(5):688-695
[40]Cockcroft DW.Mechanisms of airway hyperrespons iveness Davis BE[J].J Allergy Clin Immunol,2007;119(1):254-255
[41]李凝.神经肽和哮喘[J].国外医学呼吸系统分册,2002,22(1):45-48
[42]谢启文主编.现代神经内分泌学[M].上海:上海医科大学出版社,1999:60
[43]Stimson WH.Oestrogenandhuman Tlym Phoeytes:Preseneeofs Peeifiereee Ptors in the T-suP Pressor/eytotoxie subset[J].Seand J Immunol,1998:28:345-350
[44]宫兆华.哮喘神经内分泌免度网络紊乱及补肾益气药对其调节作用的研究(博士学位论文).上海:复旦大学,2005
[45]李明华.哮喘病的新概念和治疗研究进展[J].中国临床医生,2004,32(2):2-4
[46]Heaney LG,Robinson DS.Severe asthma treatment:need for characterizing patients[J].Lancet,2005,365(9463):974-976
[47]Bloom JW,Chaeho J,Lohman IC,et al.Diferential control of essinophil survival by glucocorticoids[J].Apoptosis,2004,9(1):97-104
[48]Scanlon PD,Connett JE,Wise RA,et al.Loss of bone density with inhaled triamcinolone in Lung Health Study Ⅱ[J].Am J Respir Crit Care Med,2004;170:1302-1309
[49]O' Connor BJ,Lofdahl CG,Balter M,et al.Zileuton added to low-dose inhaled beclomethasone for the treatment of moderate to severe persistent asthma[J]. Respir Med,2007;101:1088-1096
[50]刘自力.培土生金法对脾虚哮喘大鼠嗜酸细胞凋亡及对FasmRNA、Bcl-2mRNA表达的影响(博士学位论文).辽宁:辽宁中医药大学,2006
[51]苗明三,主编.实验动物和动物实验技术(第一版)[M].北京:中国中医药出版社,1997:100
[52]苗会,薛全福,庄逢源,等.哮喘大鼠动物模型的制备[J].基础医学与临床,1998,18(1):72-78
[53]Joseph-Bowen J,de Klerk N,Holt PG,et al.Relationship of asthma,atopy,and bronchial responsivenese to serum eosinophil cationic proteins in early childhood[J].J Allergy Clin Immunol,2004;114:1040-1042
[54]Fabbri LM,Romagnoli M,Corbetta L,et al.Differences in airway inflammation in patients with fixed airflow obstruction due to asthma or chronic obstructive pulmonary disease[J].Am J Respir Crit Care Med,2003;167:418-424
[55]Peng T,Hac L,Madri JA,et al.Role of C5 in the development of airway inflammation,airway hyperresponsiveness,and ongoing airway response[J].J Clin Invest,2005;115:1590-1595
[56]Berro AI,Perry GA,Agrawal DK.Increased expression and activation of CD30induce apoptosis in human blood cosinophils[J].J Immunol,2004;173:2174-2178
[57]郭淑文,孟庆刚,王硕仁.病证结合模型建立的思路[J].中医药学报,2001,29(1):2-4
[58]Wood N,WhittersMJ,Jacobson BA,et al.Enhanced interleukin(IL)-13responses in mice lacking IL-13 recep tor alpha 2[J].J Exp Med,2003;197(6):703-709
[59]LI Ya-fei,GUO Bo-tao,HAN Jia-xin,et al.Association between interleukin-4polymorphisms and asthma[J].Journal of Medical Colleges of PLA,2007;22(5):312-319
[60]Ngoc PL,Gold DR,TzianabesAO,et al.Cytokines,allergy,and asthma[J].Curt Op in Allergy Clin Immunol,2005;5(2):161-166
[61]Romagnani S.Cytokines and chemoattractants in allergic inflammation[J].Mol Immunol,2002;38(12-13):881-885
[62]王文建,杨莉.地塞米松对支气管哮喘模型大鼠气道重建的影响[J].郑州大学学报:医学版,2007,42(1):130-133
[63]SugitaM,Kuribayashi K,Nakagomi T,et al.Allergic bronchial asthma:airway inflammation and hyperresponsiveness[J].Intern Med,2003;42(8):636-643
[64]Shen H,O' Byrne PM,Ellis R,et al.The effects of intranasalbudesonide on allergen-induced production of interleukin-5 and eotaxin,airways,blood,and bone marrow eosinophilia,andeosinophil progenitor expansion in sensitized mice[J].Am J Resp ir Crit CareMed,2002;166(2):146-153
[65]Joos GF,PR Germonpre,JC Kips,et al.Sensory neuropeptides and the human lower airways:present state and future directions[J].Eur Respir J,1994;7:1161-1171
[66]Barnes PJ.Overview of neural mechanisms in asthma[J].Pulm Pharmacol,1995;8(4/5):151-159
[67]Spina D.Airway sensory nerves:a burning issue in asthma?[J].Thorax,1996;51(3):335-337
[68]高振,阿地力江·阿布力米提,哈木拉提·吾甫尔.哮喘的下丘脑-垂体-肾上腺轴紊乱及中医药介入的影响[J].实用医学杂志,2008,24(11):1855-1856
[69]谢启文主编.神经肽(第一版)[M].上海:复旦大学出版社.2004:164,324,331-333,344
[70]Maghni K,Michoud MC,Alles M,et al.Airway smooth muscle cells express functional neurokinin-1 receptors and the nerve-derived preprotachykinin-a gene:regulation by passive sensitization[J].Am J Respir Cell Mol Biol,2003;28(1):103-110
[71]Groneberg DA,Quarcoo D,Frossard N,et al.Neurogenic mechanisms in bronchial inflammatory diseases[J].Allergy,2004;59:1139-1152
[72]Joos GF,Germonpre PR,Kips JC,et al.Sensory neuropeptides and the human lower airways:present state and future directions[J].Eur Respir J,1994;7:1161-1171
[73]Robinson DS.The role of the mast cell in asthma:induction of airway hyperresponsiveness by interaction with smooth muscle[J]? J Allergy Clin Immunol,2004;114:58-65
[74]Opconner TM,Opconnell J,O' brien D I et al.The Role of substance P in inflammatory disease[J].J Cell Physiol,2004;201:167-180
[75]De Swert KO,Toumoy KG,Joos GF,et al.The role of the tachykinin NK1 receptor in airway changes in a mouse model of allergic asthma[J].J Allergy Clin Immunol,2004;113(6):1093-1097
[76]Groneberg DA,Quarcoo D,Frossard N et al.Neurogenic mechanisms in bronchial inflammatory diseases[J].Allergy,2004;59:1139-1152
[77]van der Kleij H P,Kraneveld A D,Redegeld F A,et al.The tachykinin NK1receptor is crucial for the development of nonatopic airway inflammation and hyperresponsiveness[J].Eur J Pharmacol,2003;476(3):249-255
[78]李淼.RT-PCR检测NK1mRNA在哮喘豚鼠气道表达的研究[J].辽宁医学杂志,2008,22(4):191-192
[79]李利生,曲瑞瑶,郭华,等.冷应激大鼠血浆及回肠P物质和VIP的变化[J].首都医科大学学报,2002,23(2):113-114
[80]张忠兵,张学庸,陈希陶,等.脾虚泄泻患者肠粘膜中SP和VIP初步探讨[J].中西医结合杂志,1991,11(3):144-146,132
[81]刘素梅,曲瑞瑶,王伟,等.实验性脾虚证大鼠胃窦部SP、VIP、CGRP含量的研究[J].首都医科大学学报,1998,19(2):105-108
[82]曲瑞瑶,刘素梅,王伟,等.实验性脾虚证大鼠胃肠动力学与胃肠肽关系的研究[J].中国中西医结合杂志,1998,18(基础理论研究特集):237-240,391
[83]刘素梅,曲瑞瑶,曾文红,等.实验性脾虚证大鼠小肠P物质、血管活性肠肽、降钙素基因相关肽含量的变化[J].中国中西医结合杂志,1997,17(基础理论研究特集):265-267,308
[84]郭华,曲瑞瑶,常延滨,等.实验性脾虚证大鼠回肠P物质和血管活性肠肽含量的变化[J].首都医科大学学报,2001,22(3):199-201
[85]李利生,曲瑞瑶,王伟,等.实验性脾虚大鼠与冷应激大鼠回肠段SP、VIP含量的变化[J].深圳中西医结合杂志,2002,12(2):75-76,79
[86]Garcia-V illar-R;Dupuis-C;M artino lle-Jpect.Functional evidence fo r NO-synthase activationby substance P th rough a mechanism no t invo lying classical tachyk in in receptors in guinea-pig ileum in vitro[J].BR-J-Pharmacal.1996;118(5):1253-1256
[87]Cayabyab-FS,J imenez-M,V ergera-Petal.Influence of nitric oxide and vasoactive intestinal pep tide on the spontaneous and triggered electrical and mechanical activities of the canine ileum[J].Can-J-Physiol-Pharmacol.1997;75(5):383-387
[88]周吕主编.胃肠生理学[M].北京:科学出版社,1991:738
[89]陈华,盛丽先.胃肠激素与脾虚型小儿厌食症关系探讨[J].中国中西医结合脾胃杂志,2000,8(3):153-154
[90]高碧珍,黄爱民,李灿东,等.不同证候胃炎模型血生化指标的实验研究[J].福建中医学院学报,2005,15(1):30-33
[91]吴娟,田德禄.慢性浅表性胃炎脾胃湿热证胃黏膜G、D细胞变化及胃泌素、生长抑素的表达[J].世界华人消化杂志,2008,16(34):3840-3843
[92]钱会南,吴海霞,王乐,等.健脾与补肾对脑内神经肽Y水平和基因表达的影响[J].中国中医药信息杂志,2006,13(4):29-31
[93]刘自力,张庆荣,吴兆利.论脾虚为哮喘宿根[J].中医药学刊,2006,24(2):294-295
[94]张庆荣,谷峰,赵世芬.脾虚对哮喘氧化抗氧化平衡改变的影响[J].辽宁中医杂志,1999,26(12):574-575
[95]孙晓玲,闫英海,张庆荣.培土生金对脾虚哮喘大鼠气道环核苷酸水平的影响[J].辽宁中医杂志,2001,28(8):507-508
[96]张庆荣,陈金,赵世芬.脾虚对哮喘大鼠嗜酸细胞凋亡的影响[J].中医药学刊,2005,23(12):2129-2130
[97]张焱,张庆荣.培土生金法对脾虚哮喘大鼠一氧化氮水平改变影响的实验研究[J].吉林中医药,2007,27(10):52-54
[98]Groneberg DA,Quarcoo D,Frossard N,et al.Neurogenic mechanisms in bronchial inflammatory diseases[J].Allergy,2004;59:1139-1141
[99]Joos GF,Germonpre PR,Pauwels RA.Neurogenic inflammation in human airways:is it important?[J].Thorax,1995;50(3):217-219
[100]俞茹云,王彤,殷凯生,等.哮喘大鼠肺内感觉神经-内分泌细胞分布和密度的变化[J].南京医科大学学报(自然科学版),2008,28(30):316-320
[101]张少军,童瑶.肺与大肠相表里的现代研究及评析[J].中医函授通讯,2000,19(3):14-16
[102]E M stermbetg.Neuroendoerine regulation of autoimmune/inflammatory disease[J].Journal of Endocrinology,2001;169:29-33
[103]陈晓萍,徐远扬.神经、内分泌、免疫网络的通用生物学语言[J].自然杂志,2002,24(4):194
[104]刘永琦,王文.虚证的免疫学本质[J].中国中医基础医学杂志,2003,9(5):7-10,13
[105]刘文俊,李荣兴.中医五脏实质是人体生命本质[J].中国中医基础医学杂志,1995,1(3):16
© 2004-2018 中国地质图书馆版权所有 京ICP备05064691号 京公网安备11010802017129号 地址：北京市海淀区学院路29号 邮编：100083 电话：办公室：(+86 10)66554848；文献借阅、咨询服务、科技查新：66554700 |