肾阳虚对消化吸收功能的影响及其机制研究
摘要
目的：本课题立足肾脾相关理论，以脾阳根于肾阳为切入点，以双侧肾上腺大部切除术建立肾阳虚证动物模型为平台，观察模型大鼠的消化吸收功能，围绕血清GAS、血清D-木糖含量，探讨细胞凋亡及凋亡相关蛋白Caspase-3、Bcl-2、Bax的表达与肾阳虚影响消化吸收功能的相关机制，并观察参附汤对肾阳虚证模型大鼠消化吸收指标的影响。运用现代研究手段检测胃窦粘膜组织、小肠粘膜吸收细胞的凋亡率，胃窦粘膜组织G细胞的表达及G细胞表达Caspase-3的百分率，小肠粘膜吸收细胞的Bcl-2、Bax表达及其超微病理形态等消化吸收相关指标，旨在从分子基因水平初步探讨肾阳虚影响消化吸收功能的可能机制。
方法：本课题采用8周龄雄性SD大鼠，以双侧肾上腺大部切除方法建立肾阳虚证动物模型，并设假手术组作为对照。术后4周，将手术大鼠随机分为模型组和参附汤组。参附汤组大鼠于每天上午8:30灌胃给予参附汤水煎液（药物浓度为每ml药液含生药0.5g）1.6ml/200g体重，假手术组和模型组均灌胃给予等容量的生理盐水。共4周。实验结束时，测定实验大鼠体温，收集24小时尿液，采集血液，留取胃窦粘膜组织及小肠粘膜组织，分离、收集小肠粘膜吸收细胞进行相关指标检测：①检测24小时尿17-OH-CS含量；②检测血清GAS、D-木糖含量；③观察胃窦粘膜组织、小肠粘膜组织显微结构改变；④观察小肠粘膜吸收细胞的超微结构改变；⑤TUNEL法检测胃窦粘膜组织凋亡率、免疫组化法检测G细胞表达的平均光密度值及免疫组化双染法检测G细胞表达Caspase-3的百分率；⑥取小肠粘膜吸收细胞单细胞悬液，用流式细胞仪检测小肠粘膜吸收细胞的凋亡率；⑦免疫组化法检测小肠粘膜吸收细胞的Bcl-2、Bax蛋白表达的平均光密度。
结果：①体温、24小时尿液指标检测结果：与假手术组比较，模型组大鼠体温、尿17-OH-CS含量显著降低（P＜0.01）;与模型组比较，参附汤组大鼠体温、尿17-OH-CS含量均显著升高（P＜0.01）。
②血清指标检测结果：与假手术组比较，模型组大鼠血清GAS、D-木糖含量均显著降低（P＜0.01）；与模型组比较，参附汤组大鼠血清GAS、D-木糖含量均显著升高（P＜0.01）。
③胃窦粘膜组织指标检测结果：与假手术组比较，模型组大鼠的胃窦粘膜组织变薄，细胞间隙变大，粘膜腺体有不同程度的排列紊乱、疏松、出现大量空泡样变；与模型组比较，参附汤组大鼠的胃窦粘膜组织稍厚，细胞间隙变小，腺细胞排列较规则，空泡样变减少。与假手术组比较，模型组大鼠胃窦粘膜组织的凋亡率、G细胞表达Caspase-3的百分率显著升高（P＜0.01），G细胞表达的平均光密度值显著降低（P＜0.01）；与模型组比较，参附汤组大鼠胃窦粘膜组织的凋亡率、G细胞表达Caspase-3的百分率显著下降（P＜0.01），G细胞表达的平均光密度值显著升高（P＜0.01）。
④小肠粘膜组织指标检测结果：光镜结果显示，与假手术组比较，模型组大鼠小肠粘膜组织厚度略欠均匀，绒毛边缘不整齐，部分上皮细胞脱落、缺失；与模型组比较，参附汤组大鼠小肠粘膜组织厚度较均匀，绒毛排列规则，上皮基本完整。电镜结果显示，与假手术组比较，模型组大鼠小肠粘膜吸收细胞微绒毛略显稀疏，细胞间连接疏松，细胞器模糊，细胞核膜内陷，异染色质凝聚、边集，胞质内空泡多见；参附汤组大鼠小肠粘膜吸收细胞超微结构较模型组情况有显著改善，细胞间连接比较紧密，胞质内空泡化结构少见，异染色质凝集、边集及核膜内陷现象均较模型组好转。免疫组化结果显示，与假手术组比较，模型组大鼠小肠粘膜组织吸收细胞Bcl-2、Bax蛋白表达均显著升高（P＜0.01）；与模型组比较，参附汤组大鼠小肠粘膜组织吸收细胞Bcl-2蛋白的表达显著升高（P＜0.01），Bax蛋白的表达显著降低（P＜0.01）。
⑤小肠粘膜吸收细胞指标检测结果：与假手术组比较，模型组大鼠小肠粘膜组织吸收细胞的凋亡率显著升高（P＜0.01）；与模型组比较，参附汤组大鼠小肠粘膜组织吸收细胞的凋亡率显著降低（P＜0.01）。
结论：①采用肾上腺大部切除法建立肾阳虚证大鼠模型，模型组大鼠的一般表现符合肾阳虚的诊断标准，且模型组大鼠体温、24小时尿17-OH-CS含量均显著下降，说明肾阳虚证动物模型建立成功。
②模型组大鼠血清GAS、D-木糖含量较假手术组显著下降，说明肾阳虚证模型动物的消化吸收功能下降。
③模型组大鼠胃窦粘膜组织的凋亡率、G细胞表达Caspase-3的百分率显著升高，G细胞表达的平均光密度值显著降低，说明胃窦粘膜组织及其G细胞的过度凋亡影响了GAS的分泌，进而影响了模型大鼠的消化功能。
④模型组大鼠小肠粘膜吸收细胞的超微组织结构改变符合凋亡细胞的典型表现，吸收细胞的凋亡率明显升高，同时，小肠吸收细胞Bax蛋白的表达显著升高，凋亡明显，凋亡与增殖失衡，使小肠粘膜吸收细胞凋亡增加，增殖不足，得不到及时更新，从而影响了模型大鼠的吸收功能。
⑤温补肾阳方药—参附汤可以调节凋亡相关蛋白Caspase-3、Bcl-2、Bax的表达，从而抑制消化吸收相关组织细胞的过度凋亡，促进其更新，维护其功能，从而改善肾阳虚证模型大鼠的消化吸收功能。
本课题首次采用双侧肾上腺大部切除的方法建立肾阳虚证动物模型,率先运用现代技术方法研究肾阳虚影响消化吸收功能的相关机制。肾阳虚证模型大鼠胃窦粘膜组织G细胞凋亡关键蛋白Caspase-3的表达显著增强，小肠粘膜吸收细胞的促凋亡基因Bax表达显著增强，使模型大鼠消化吸收相关组织细胞凋亡增加，功能受损，进而表现为消化吸收功能下降。温补肾阳方药—参附汤可以下调胃窦粘膜组织G细胞的Caspase-3表达、小肠粘膜吸收细胞的Bax表达，抑制过度凋亡，改善肾阳虚导致的消化吸收功能下降。由此可知，消化吸收相关组织细胞的过度凋亡可能是肾阳虚影响消化吸收功能的机制之一。
Objective: The kidney-Yang deficiency rat model were set by the bilateraladrenal subtotal excision to observe the digestive and absorptive function inmodel rats based on the related theory of spleen and kidney, with spleen yangrooting in kidney-yang as the breakthrough point. Around the serum content ofGAS and D-xylose，the related mechanism was also investigated between theexpression of apoptosis and apoptosis related protein (such as Caspase-3,Bcl-2, Bax) and the digestive and absorptive function effected by kidney-Yangdeficiency. Else, the effect on digestive and absorptive index was observed inkidney-yang deficiency model rat treated by Shenfu Decoction. Modernresearch methods were used to detect the digestive and absorptive indexs,incuding the rate of apoptosis of gastric antrum mucosa tissue and intestinalmucosal absorptive epithelial cell, the expression in gastric antrum mucosatissue’s G cells, the percentage of Caspase-3expressed by G cells, theexpression of Bcl-2and Bax, and ultrastructural pathologic morphology inintestinal mucosal absorptive epithelial cell, et al. Our research aimed toinvestigate the mechanism of digestive and absorptive function effected bykidneyYang deficiency from Molecular gene level.
Methods: Male8weeks aged SD rats were adopted，which resected bilateraladrenal subtotal to set up kidney-yang deficiency model, and which withoutresected bilateral adrenal were as sham operation group.4weeks afteroperation, the operative rats were randomly divided into model group andShenfu Decoction group. In every morning8:30, every rat in Shenfu Decoction group was given Suspension liquid of1.6ml/200g ShenfuDecoction through the lavage, and this drug concentrations for each of1mlsolution contained0.5g crude drugs. Every rat in Sham operation group andmodel group both had been given the physiological saline of the same capacityfor4weeks.We determinated the temperature of all experimental rats,collected their blood and24h urine at the end of the experiment. Gastricantrum mucosa tissue and intestinal mucosa tissue were left to test relatedindexes, by separating and collecting intestinal mucosal absorptive epithelialcell:①To detect the content of24hour urinary17-OH-CS;②To detect thecontent of serum GAS and D-xylose;③To observe the microstructurechanges of gastric antrum mucosa tissue and intestinal mucosa tissue;④Toobserve the ultrastructural changes in absorptive epithelial cell of intestinalmucosa tissue;⑤To detect the apoptosis rate of gastric antrum mucosa tissueby TUNEL, the average optical density value of G cells byimmunohistochemistry and the percentage of Caspase-3expressed in G cellsthrough immunohistochemical double staining method;⑥Taking the singlecell suspension of intestinal mucosal tissue absorptive epithelial cells, to detectthe apoptosis rate of absorptive epithelial cell of intestinal mucosa tissue byflow cytometry;⑦To examined the average optical density value of Bcl-2and Bax in intestinal mucosal tissue absorptive epithelial cells byimmunohistochemistry.
Results:①Detection of temperature and24hour urinary index: Comparedwith the sham operation group, the temperature and content of urine17-OH-CS of the model group decreased significantly(P<0.01); Comparedwith the model group, the temperature and content of urine17-OH-CS of theShenfu Decoction group increased significantly(P<0.01).
②Detection of serum index: Compared with the sham operation group,the content of serum GAS and D-xylose of the model group decreased significantly(P<0.01); Compared with the model group, the content of serumGAS and D-xylose of the Shenfu Decoction group increased significantly(P<0.01).
③Detection of gastric antrum mucosa tissue index: The results showedthat of the light microscope：Compared with the sham operation group, theantrum mucosa tissue of the model group became thiner, the cell gap becamelarger. Else, the mucous glands have different degrees of alignment disorders,loose；the emergence of a large number of vacuolar degeneration. Comparedwith the model group, the antrum mucosa tissue of the Shenfu Decoctiongroup was slightly thick, the cell gap became smaller, the mucous glands cellsarranged more regulation, and the vacuolar degeneration had reduced. TUNELand immunohistochemistry results show：Compared with the sham operationgroup, the apoptosis rate of gastric antrum mucosa tissue and the percentage ofCaspase-3expressed in G cells of the model group increased significantly(P<0.01), while the average optical density value of G cells decreasedsignificantly(P<0.01). Compared with the model group, the apoptosis rate ofgastric antrum mucosa tissue and the percentage of Caspase-3expressed in Gcells of the Shenfu Decoction group decreased significantly(P<0.01), whilethe average optical density value of G cells increased significantly(P<0.01).
④Detection of intestinal mucosa tissue index: The results showed that ofthe light microscope：Compared with the sham operation group, intestinalmucosa tissue biopsy of the model group showed that mucosaangl thicknesswas less uniform. And the villus edge was irregular, partial shedding of theepithelium fell off and deleted. Compared with the model group, intestinalmucosal thickness of the Shenfu Decoction group was more uniform, thevillus edge was more regular, and intestinal mucosal epithelial tissue was morecomplete. The results showed that of the electron microscope: Compared withthe sham operation group, electron microscopy showed that intestinal epithelial cell microvilli slightly was a little sparse, and junction between cellswas loose. Thus that cell device was fuzzy in cell membrane invaginating. Atthe same time, heterochro-matin of this group had a condensation and frontierset, and cytoplasmic had many vacuoles. However, ultrastructural of intestinalepithelial cells of the Shenfu Decoction group had remarkable improvementcompared with the model group. Compared with the sham operation group,immunohistochemistry result showed that the expression of Bcl-2and Bax inintestinal mucosal absorptive epithelial cell of the model group increasedsignificantly (P<0.01). Compared with the model group, the expression ofBcl-2in intestinal mucosal absorptive epithelial cell of the Shenfu Decoctiongroup increased significantly (P<0.01), while the expression of Bax decreasedsignificantly (P<0.01).
⑤Detection of intestinal absorption cell index: Compared with the shamoperation group, the apoptosis rate of intestinal mucosa absorptive epithelialcells of the model group increased significantly (P<0.01). Compared with themodel group, the apoptosis rate of intestinal mucosa absorptive epithelial cellsof the Shenfu Decoction group decreased significantly (P<0.01).
Conclusion:①The model rats with deficiency of kidney-yang wasestablished by adrece which conformed to the diagnostic criteria of deficiencyof kidney-yang. Thus the temperature and24h urinary17-OH-CS content ofthe model group rats were significantly decreased. So it suggests that thereplication of this animal model was successful.
②Compared with the sham operation group, the content of serum GASand D-xylose of the model group decreased significantly. This result illustratesthat the digestion and absorption function of kidney-yang deficiency animalhad a decline.
③The apoptosis rate of gastric antrum mucosa tissue and the percentageof Caspase-3expressed in G cells of the model group increased significantly, while the average optical density value of G cells decreased significantly. So itsuggests that the abnormal apoptosis of gastric antrum mucosa tissue and its Gcell has a influence on secretion of GAS.
④Ultrastructural organization structure in intestine absorptive epithelialcells of the model group conformed to the typical features of apoptosis cell.The apoptosis rate of intestinal mucosa absorptive epithelial cells of the modelgroup increased significantly. At the same time, the expression of bax had asignificant increase. Thus there had a remarkable apoptosis in the model rats,and had a unbalance between proliferation and apoptosis. Then the apoptosisof intestinal mucosa absorptive epithelial cell increased. However theproliferation was insufficient which could't be updated in time. Finally it had anegative effect on absorption function of the model rats.
⑤Shenfu Decoction, a prescription for warming and recuperating thekidney-yang, can inhibit the excessive apoptosis of correlated tissue cells byadjusting the expression of Caspas-3、Bcl-2and Bax. Else this drug can alsopromote the cell renewal, maintenance of its function, thereby improving thedigestive and absorptive function of kidney-yang deficiency model rats.
Our research first uses the bilateral adrenal subtotal excision method toestablish the kidney-yang deficiency animal model. We take the lead in the useof modern technology and method to study the mechanism of digestive andabsorptive function affected by kidney-yang deficiency.As the key protein ofapoptosis in G-cell of gastric antrum mucosa tissue, Caspase-3expression ofthe kidney-yang deficiency model rats increased remarkably. So did theexpression of Bax which was the pro-apoptotic gene in intestinal mucosalabsorptive epithelial cell. It increased the apoptosis of digestive and absorptiverelated tissue cell in the model rats. Then the performance for the digestiveand absorptive function declined. Shenfu Decoction, a prescription forwarming and recuperating the kidney-yang, can improve the impaired digestive and absorptive function leaded by kidney-yang deficiency throughdecreasing the express of Caspase-3in G cells of gastric antrum mucosa tissue,and Bax in intestinal mucosal absorptive epithelial cell. Thus, abnormalapoptosis may be one of the mechanism that kidney-yang deficiency can havea negative effect on digestive and absorptive function.
方法：本课题采用8周龄雄性SD大鼠，以双侧肾上腺大部切除方法建立肾阳虚证动物模型，并设假手术组作为对照。术后4周，将手术大鼠随机分为模型组和参附汤组。参附汤组大鼠于每天上午8:30灌胃给予参附汤水煎液（药物浓度为每ml药液含生药0.5g）1.6ml/200g体重，假手术组和模型组均灌胃给予等容量的生理盐水。共4周。实验结束时，测定实验大鼠体温，收集24小时尿液，采集血液，留取胃窦粘膜组织及小肠粘膜组织，分离、收集小肠粘膜吸收细胞进行相关指标检测：①检测24小时尿17-OH-CS含量；②检测血清GAS、D-木糖含量；③观察胃窦粘膜组织、小肠粘膜组织显微结构改变；④观察小肠粘膜吸收细胞的超微结构改变；⑤TUNEL法检测胃窦粘膜组织凋亡率、免疫组化法检测G细胞表达的平均光密度值及免疫组化双染法检测G细胞表达Caspase-3的百分率；⑥取小肠粘膜吸收细胞单细胞悬液，用流式细胞仪检测小肠粘膜吸收细胞的凋亡率；⑦免疫组化法检测小肠粘膜吸收细胞的Bcl-2、Bax蛋白表达的平均光密度。
结果：①体温、24小时尿液指标检测结果：与假手术组比较，模型组大鼠体温、尿17-OH-CS含量显著降低（P＜0.01）;与模型组比较，参附汤组大鼠体温、尿17-OH-CS含量均显著升高（P＜0.01）。
②血清指标检测结果：与假手术组比较，模型组大鼠血清GAS、D-木糖含量均显著降低（P＜0.01）；与模型组比较，参附汤组大鼠血清GAS、D-木糖含量均显著升高（P＜0.01）。
③胃窦粘膜组织指标检测结果：与假手术组比较，模型组大鼠的胃窦粘膜组织变薄，细胞间隙变大，粘膜腺体有不同程度的排列紊乱、疏松、出现大量空泡样变；与模型组比较，参附汤组大鼠的胃窦粘膜组织稍厚，细胞间隙变小，腺细胞排列较规则，空泡样变减少。与假手术组比较，模型组大鼠胃窦粘膜组织的凋亡率、G细胞表达Caspase-3的百分率显著升高（P＜0.01），G细胞表达的平均光密度值显著降低（P＜0.01）；与模型组比较，参附汤组大鼠胃窦粘膜组织的凋亡率、G细胞表达Caspase-3的百分率显著下降（P＜0.01），G细胞表达的平均光密度值显著升高（P＜0.01）。
④小肠粘膜组织指标检测结果：光镜结果显示，与假手术组比较，模型组大鼠小肠粘膜组织厚度略欠均匀，绒毛边缘不整齐，部分上皮细胞脱落、缺失；与模型组比较，参附汤组大鼠小肠粘膜组织厚度较均匀，绒毛排列规则，上皮基本完整。电镜结果显示，与假手术组比较，模型组大鼠小肠粘膜吸收细胞微绒毛略显稀疏，细胞间连接疏松，细胞器模糊，细胞核膜内陷，异染色质凝聚、边集，胞质内空泡多见；参附汤组大鼠小肠粘膜吸收细胞超微结构较模型组情况有显著改善，细胞间连接比较紧密，胞质内空泡化结构少见，异染色质凝集、边集及核膜内陷现象均较模型组好转。免疫组化结果显示，与假手术组比较，模型组大鼠小肠粘膜组织吸收细胞Bcl-2、Bax蛋白表达均显著升高（P＜0.01）；与模型组比较，参附汤组大鼠小肠粘膜组织吸收细胞Bcl-2蛋白的表达显著升高（P＜0.01），Bax蛋白的表达显著降低（P＜0.01）。
⑤小肠粘膜吸收细胞指标检测结果：与假手术组比较，模型组大鼠小肠粘膜组织吸收细胞的凋亡率显著升高（P＜0.01）；与模型组比较，参附汤组大鼠小肠粘膜组织吸收细胞的凋亡率显著降低（P＜0.01）。
结论：①采用肾上腺大部切除法建立肾阳虚证大鼠模型，模型组大鼠的一般表现符合肾阳虚的诊断标准，且模型组大鼠体温、24小时尿17-OH-CS含量均显著下降，说明肾阳虚证动物模型建立成功。
②模型组大鼠血清GAS、D-木糖含量较假手术组显著下降，说明肾阳虚证模型动物的消化吸收功能下降。
③模型组大鼠胃窦粘膜组织的凋亡率、G细胞表达Caspase-3的百分率显著升高，G细胞表达的平均光密度值显著降低，说明胃窦粘膜组织及其G细胞的过度凋亡影响了GAS的分泌，进而影响了模型大鼠的消化功能。
④模型组大鼠小肠粘膜吸收细胞的超微组织结构改变符合凋亡细胞的典型表现，吸收细胞的凋亡率明显升高，同时，小肠吸收细胞Bax蛋白的表达显著升高，凋亡明显，凋亡与增殖失衡，使小肠粘膜吸收细胞凋亡增加，增殖不足，得不到及时更新，从而影响了模型大鼠的吸收功能。
⑤温补肾阳方药—参附汤可以调节凋亡相关蛋白Caspase-3、Bcl-2、Bax的表达，从而抑制消化吸收相关组织细胞的过度凋亡，促进其更新，维护其功能，从而改善肾阳虚证模型大鼠的消化吸收功能。
本课题首次采用双侧肾上腺大部切除的方法建立肾阳虚证动物模型,率先运用现代技术方法研究肾阳虚影响消化吸收功能的相关机制。肾阳虚证模型大鼠胃窦粘膜组织G细胞凋亡关键蛋白Caspase-3的表达显著增强，小肠粘膜吸收细胞的促凋亡基因Bax表达显著增强，使模型大鼠消化吸收相关组织细胞凋亡增加，功能受损，进而表现为消化吸收功能下降。温补肾阳方药—参附汤可以下调胃窦粘膜组织G细胞的Caspase-3表达、小肠粘膜吸收细胞的Bax表达，抑制过度凋亡，改善肾阳虚导致的消化吸收功能下降。由此可知，消化吸收相关组织细胞的过度凋亡可能是肾阳虚影响消化吸收功能的机制之一。
Objective: The kidney-Yang deficiency rat model were set by the bilateraladrenal subtotal excision to observe the digestive and absorptive function inmodel rats based on the related theory of spleen and kidney, with spleen yangrooting in kidney-yang as the breakthrough point. Around the serum content ofGAS and D-xylose，the related mechanism was also investigated between theexpression of apoptosis and apoptosis related protein (such as Caspase-3,Bcl-2, Bax) and the digestive and absorptive function effected by kidney-Yangdeficiency. Else, the effect on digestive and absorptive index was observed inkidney-yang deficiency model rat treated by Shenfu Decoction. Modernresearch methods were used to detect the digestive and absorptive indexs,incuding the rate of apoptosis of gastric antrum mucosa tissue and intestinalmucosal absorptive epithelial cell, the expression in gastric antrum mucosatissue’s G cells, the percentage of Caspase-3expressed by G cells, theexpression of Bcl-2and Bax, and ultrastructural pathologic morphology inintestinal mucosal absorptive epithelial cell, et al. Our research aimed toinvestigate the mechanism of digestive and absorptive function effected bykidneyYang deficiency from Molecular gene level.
Methods: Male8weeks aged SD rats were adopted，which resected bilateraladrenal subtotal to set up kidney-yang deficiency model, and which withoutresected bilateral adrenal were as sham operation group.4weeks afteroperation, the operative rats were randomly divided into model group andShenfu Decoction group. In every morning8:30, every rat in Shenfu Decoction group was given Suspension liquid of1.6ml/200g ShenfuDecoction through the lavage, and this drug concentrations for each of1mlsolution contained0.5g crude drugs. Every rat in Sham operation group andmodel group both had been given the physiological saline of the same capacityfor4weeks.We determinated the temperature of all experimental rats,collected their blood and24h urine at the end of the experiment. Gastricantrum mucosa tissue and intestinal mucosa tissue were left to test relatedindexes, by separating and collecting intestinal mucosal absorptive epithelialcell:①To detect the content of24hour urinary17-OH-CS;②To detect thecontent of serum GAS and D-xylose;③To observe the microstructurechanges of gastric antrum mucosa tissue and intestinal mucosa tissue;④Toobserve the ultrastructural changes in absorptive epithelial cell of intestinalmucosa tissue;⑤To detect the apoptosis rate of gastric antrum mucosa tissueby TUNEL, the average optical density value of G cells byimmunohistochemistry and the percentage of Caspase-3expressed in G cellsthrough immunohistochemical double staining method;⑥Taking the singlecell suspension of intestinal mucosal tissue absorptive epithelial cells, to detectthe apoptosis rate of absorptive epithelial cell of intestinal mucosa tissue byflow cytometry;⑦To examined the average optical density value of Bcl-2and Bax in intestinal mucosal tissue absorptive epithelial cells byimmunohistochemistry.
Results:①Detection of temperature and24hour urinary index: Comparedwith the sham operation group, the temperature and content of urine17-OH-CS of the model group decreased significantly(P<0.01); Comparedwith the model group, the temperature and content of urine17-OH-CS of theShenfu Decoction group increased significantly(P<0.01).
②Detection of serum index: Compared with the sham operation group,the content of serum GAS and D-xylose of the model group decreased significantly(P<0.01); Compared with the model group, the content of serumGAS and D-xylose of the Shenfu Decoction group increased significantly(P<0.01).
③Detection of gastric antrum mucosa tissue index: The results showedthat of the light microscope：Compared with the sham operation group, theantrum mucosa tissue of the model group became thiner, the cell gap becamelarger. Else, the mucous glands have different degrees of alignment disorders,loose；the emergence of a large number of vacuolar degeneration. Comparedwith the model group, the antrum mucosa tissue of the Shenfu Decoctiongroup was slightly thick, the cell gap became smaller, the mucous glands cellsarranged more regulation, and the vacuolar degeneration had reduced. TUNELand immunohistochemistry results show：Compared with the sham operationgroup, the apoptosis rate of gastric antrum mucosa tissue and the percentage ofCaspase-3expressed in G cells of the model group increased significantly(P<0.01), while the average optical density value of G cells decreasedsignificantly(P<0.01). Compared with the model group, the apoptosis rate ofgastric antrum mucosa tissue and the percentage of Caspase-3expressed in Gcells of the Shenfu Decoction group decreased significantly(P<0.01), whilethe average optical density value of G cells increased significantly(P<0.01).
④Detection of intestinal mucosa tissue index: The results showed that ofthe light microscope：Compared with the sham operation group, intestinalmucosa tissue biopsy of the model group showed that mucosaangl thicknesswas less uniform. And the villus edge was irregular, partial shedding of theepithelium fell off and deleted. Compared with the model group, intestinalmucosal thickness of the Shenfu Decoction group was more uniform, thevillus edge was more regular, and intestinal mucosal epithelial tissue was morecomplete. The results showed that of the electron microscope: Compared withthe sham operation group, electron microscopy showed that intestinal epithelial cell microvilli slightly was a little sparse, and junction between cellswas loose. Thus that cell device was fuzzy in cell membrane invaginating. Atthe same time, heterochro-matin of this group had a condensation and frontierset, and cytoplasmic had many vacuoles. However, ultrastructural of intestinalepithelial cells of the Shenfu Decoction group had remarkable improvementcompared with the model group. Compared with the sham operation group,immunohistochemistry result showed that the expression of Bcl-2and Bax inintestinal mucosal absorptive epithelial cell of the model group increasedsignificantly (P<0.01). Compared with the model group, the expression ofBcl-2in intestinal mucosal absorptive epithelial cell of the Shenfu Decoctiongroup increased significantly (P<0.01), while the expression of Bax decreasedsignificantly (P<0.01).
⑤Detection of intestinal absorption cell index: Compared with the shamoperation group, the apoptosis rate of intestinal mucosa absorptive epithelialcells of the model group increased significantly (P<0.01). Compared with themodel group, the apoptosis rate of intestinal mucosa absorptive epithelial cellsof the Shenfu Decoction group decreased significantly (P<0.01).
Conclusion:①The model rats with deficiency of kidney-yang wasestablished by adrece which conformed to the diagnostic criteria of deficiencyof kidney-yang. Thus the temperature and24h urinary17-OH-CS content ofthe model group rats were significantly decreased. So it suggests that thereplication of this animal model was successful.
②Compared with the sham operation group, the content of serum GASand D-xylose of the model group decreased significantly. This result illustratesthat the digestion and absorption function of kidney-yang deficiency animalhad a decline.
③The apoptosis rate of gastric antrum mucosa tissue and the percentageof Caspase-3expressed in G cells of the model group increased significantly, while the average optical density value of G cells decreased significantly. So itsuggests that the abnormal apoptosis of gastric antrum mucosa tissue and its Gcell has a influence on secretion of GAS.
④Ultrastructural organization structure in intestine absorptive epithelialcells of the model group conformed to the typical features of apoptosis cell.The apoptosis rate of intestinal mucosa absorptive epithelial cells of the modelgroup increased significantly. At the same time, the expression of bax had asignificant increase. Thus there had a remarkable apoptosis in the model rats,and had a unbalance between proliferation and apoptosis. Then the apoptosisof intestinal mucosa absorptive epithelial cell increased. However theproliferation was insufficient which could't be updated in time. Finally it had anegative effect on absorption function of the model rats.
⑤Shenfu Decoction, a prescription for warming and recuperating thekidney-yang, can inhibit the excessive apoptosis of correlated tissue cells byadjusting the expression of Caspas-3、Bcl-2and Bax. Else this drug can alsopromote the cell renewal, maintenance of its function, thereby improving thedigestive and absorptive function of kidney-yang deficiency model rats.
Our research first uses the bilateral adrenal subtotal excision method toestablish the kidney-yang deficiency animal model. We take the lead in the useof modern technology and method to study the mechanism of digestive andabsorptive function affected by kidney-yang deficiency.As the key protein ofapoptosis in G-cell of gastric antrum mucosa tissue, Caspase-3expression ofthe kidney-yang deficiency model rats increased remarkably. So did theexpression of Bax which was the pro-apoptotic gene in intestinal mucosalabsorptive epithelial cell. It increased the apoptosis of digestive and absorptiverelated tissue cell in the model rats. Then the performance for the digestiveand absorptive function declined. Shenfu Decoction, a prescription forwarming and recuperating the kidney-yang, can improve the impaired digestive and absorptive function leaded by kidney-yang deficiency throughdecreasing the express of Caspase-3in G cells of gastric antrum mucosa tissue,and Bax in intestinal mucosal absorptive epithelial cell. Thus, abnormalapoptosis may be one of the mechanism that kidney-yang deficiency can havea negative effect on digestive and absorptive function.
引文
[1]郎庆波.肾主命门论[J].江苏中医,2001,22(1):3-4.
[2]吴敦序.中医基础理论[M].第1版.上海:上海科学技术出版社,2000:59.
[3]易青.再论“肾主水液”之内涵[J].湖北中医学院学报,2006,8(3):40-41.
[4]麻丽萍,易杰.论中医五脏生血[J].辽宁中医药大学学报,2009,11(8):5-6.
[5]吴志奎.肾生髓、髓生血理论与治疗地中海贫血的临床实践[J].中医杂志,2008,49(2):170-171.
[6]林殷,鲁兆麟.“补火生土”考辨[J].北京中医药大学学报,2003,26(3):11-13.
[7]蔡茂玉.脾主运化浅议[J].云南中医中药杂志,2004,25(3):58-59.
[1]戴芹,王怡,曲晓璐.温肾化痰方对早中期脾肾阳虚型慢性肾功能不全患者肾功能及血脂的影响[J].中国中西医结合杂志,2012,32(2):188-190.
[2]任可,马玲玲,刘美奇,等.血液透析后脾肾阳虚型患者血氧饱和度变化的临床观察[J].北京中医药大学学报（中医临床版）,2012,19(1):31-32.
[3]王晓英,苗得雨,裴妙荣.复合式造大鼠脾肾阳虚模型[J].山西中医,2012,28(7):43-46.
[4]林传权,王桂香,李茹柳,等.慢性浅表性胃炎脾虚证消化吸收障碍亚型的临床分析[J].广州中医药大学学报,2008,25(6):488-491.
[5]蔡茂玉.脾主运化浅议[J].云南中医中药杂志,2004,25(3):57-58.
[1]陈英华,欧阳轶强,孙琪,等.肾阳虚证动物模型规范化研究诊断指标选择的初步探讨[J].中国中医基础医学杂志,2003,9(10):26-34.
[2]李广曦.肾阳虚证动物模型的造模方法及其相关指标回顾[J].中国中医基础医学杂志,2000,6(4):46-55.
[3]陈小野,邹世洁,王震,等.大鼠肾上腺次全切除肾阳虚模型胃、舌病理观察[J].中国中医基础医学杂志,1999,5(11):20-23.
[4]苗明三.实验动物和动物实验技术[M].第1版.北京:中国中医药出版社,2003:242.
[5]贺石林,王键,王净净.中医科研设计与统计学[M].第1版.长沙:湖南科学技术出版社,2005:48-49.
[6]沈自尹,王文健.中医虚证辨证参考标准[J].中西医结合杂志，1986,6(10):598.
[1]邝安堃.中西医结合研究丛书—虚证研究[M].第1版.上海:上海科学技术出版社,1991:118.
[2]呼延和,张君.开郁健脾法对脾虚大鼠血清胃泌素、尿D-木糖影响的实验研究[J].中医儿科杂志,2010,6(3):12-16.
[3]李晓霞.脾气虚与Na+-K+-ATP酶活力观察[J].中国医学报,1996,11(2):83.
[4]Buts JP，Morin CL，Roy CC，et al. One-hour blood xylose test: a reliable index ofsmall bowel function [J].The Journal of Pediatries,1978,90(5):729-733.
[5]Yamamoto O, Matsunaga Y, Shiba Y,et al. Inhibition of motilin-induced phase Ⅲcontraction by pentagastrin in Heidenhain pouch dogs [J]. J Pharmacol ExpTher,1994,271:1471-1476.
[6]郑小伟,王颖,宋红.补中益气汤对脾气虚证大鼠血清胃泌素影响的实验研究[J].中华中医药杂志(原中国医药学报),2006,21(7):393-394.
[7]郑小伟,宋红,包素珍,等.实验性脾气虚大鼠胃泌素的基因表达及四君子汤的干预作用[J].中华中医药杂志(原中国医药学报),2008,23(3):264-266.
[1]王满贵,宋志宙.HE染色切片的技术操作规范[J].大同医学专科学校学报2004,(2):21.
[2]廉洁,张翠香,金明顺.HE染色的组织切片褪色的原因分析[J].齐齐哈尔医学院学报,2001,22(12):1432.
[3]蔡晓雯,沈武成.石蜡切片HE染色的体会[J].医药论坛杂志,2011,32(1):167-168.
[1]夏朝霞,何向蕾.原位细胞凋亡技术TUNEL法的改进[J].现代实用医学,2010,22(6):642-643.
[2]郭晓红,刘立新.Annexin V/PI流式细胞分析法和TUNEL法检测肝细胞凋亡的对比研究[J].山西医科大学学报[J].2008,39(5):476-479.
[3]李彦群,于凤玲,陈文雪.凋亡细胞TUNEL技术经验与改进[J].中国误诊学杂志,2001(1卷第10):1582-1583.
[4]董小莉,谭宁,符永恒,等.TUNEL法原位检测心肌细胞凋亡的改进.中国病理生理杂志,2010,26(5):1038-1040.
[5]张鸣号,彭亮,曹军.显微图像分析法与人工计数法在免疫组化结果判读中的应用[J].宁夏医科大学学报,2009,4(31):261-262.
[6]曹雪涛.免疫学技术及其应用[M].第1版.北京:科学出版社,2010:626.
[7]王伯沄,王文勇.免疫组织化学的新进展[J].中华医学会病理学分会学术年会论文汇编,2009:181.
[8]王秋生,马世文.免疫组化技术的应用[J].安庆医学,2004,25(1):37-38.
[9]许力,吴珊,杨清海,等.免疫组化技术(一)[J].诊断病理学杂志,2009,16,(2):158.
[10]石善溶.基于抗原修复技术的定量免疫组化:从实验、假说到研究计划[J].临床与实病理学杂志,2011,27(12):1271-1275.
[11]陈勤,周君,胡冠宇.石蜡切片免疫组化过程中存在的问题及解决方法[J].安徽大学学报(自然科学版),2009,33(2):85-88.
[12]高美钦,袁芳平,黄爱民,等.弹力纤维染色与免疫组织化学双重染色技术[J].中华病理学杂志,2003,32(2):176-177.
[13]应李雄,丁伟.DNA染色与免疫组织化学双重染色技术应用[J].中国组织化学与细胞化学杂志,2003,12(2):226-227.
[14]肖立,陆孝禹.CK34BE12和PSA顺序免疫组化双染色在前列腺病变中的表达[J].老年医学与保健,2003,9(1):14-18.
[15]陈罡,罗殿中,韦康来,等.双重免度组化染色法的操作体会[J].实用医技杂志,2004,11(5)下半月版:721-722.
[16]黄莺,王康敏,康安静,等.bFGF与Ki-67双重免疫组化染色法[J].诊断病理学杂志,2002,9(3):187.
[17]弓娟琴,陈志强,李文忠,等.Fas介导的凋亡与Caspase家族[J].国外医学:肿瘤学分册,2001,27(5):279-280．
[18]Stepien A, lzdebska M, Grzanka A. The types of cell death [J]. Postepy Hig MedDosw (Online),2007,61:420-428．
[19]Cryns V, Yuan J．Protease to die for [J]. Genes Dev,1998,12(11):1551-1570．
[20]赵霞,潘华峰,鞠晓云,等.胃痞消对CAG脾虚大鼠胃粘膜上皮细胞凋亡及调控蛋白Caspase-3影响的研究[J].中国中医基础医学杂志,2007,13(1):42-43.
[1]牛建昭.组织学与胚胎学[M].第1版.北京:人民卫生出版社,2002:141-143.
[2] Desoignie R,Sellin J. Propionate-initialed changes in intra-cellular pH in rabbitcoloncytes.Gastroenterology,1994,107(2):347-356.
[3]王辉,高颖.应用流式细胞仪的常见问题及解决对策[J].现代仪器,2012,18(1):64-65.
[4]胡凡.流式细胞仪在医学检验中的应用[J].中国保健营养,2012,(5):1139-1140.
[5] Brittan M, Wright NA. Stem cell in gastrointestinal structure and neoplasticdevelopment[J]. Gut,2004,53(6):899-910.
[6] Zhang C, Sheng ZY, Hu S, et al.The influence of apoptosis of mucosal epithelialcells on intestinal barrier integrity after scald in rats[J].Burns,2002,28(8):731-737.
[1]牛建昭.组织学与胚胎学[M].第1版.北京:人民卫生出版社,2002:141-143.
[2]冯新玲.恐伤孕鼠对其仔鼠脑发育的影响及其机理研究[D].湖北中医学院博士学位论文,2006:24.
[3] Otsuki Y. Various methods of apoptosis detection [J]. Acta HistochemCytochem,2000,33(4):235-241.
[4]Kalimichenko SG, Matveeva NY. Morphological characteristics of apoptosis and itssignificance in neurogenesis [J].Neurosci Behav Physiol,2008,38(4):333-344.
[5]Patel T, Gores JG. Apoptosis and hepatobiliary disease[J].Hepatol,1995,21(6):1725-1741.
[6]顾瑜蓉,李华伟,张重华,等.嗅球摘除后嗅觉神经元凋亡的实验研究[J].中华耳鼻咽喉头颈外科杂志,2006,41(4):297-300.
[7]沈强,俞彰,法京.电镜下几种凋亡细胞的形态特征[J].复旦学报(医学版),2010,37(3):322-325.
[8]Tilly JL, Kowalski KI, Johnson AL,et al. Involvement of apoptosis in ovarianfollicular atresia and postovulatory regression[J].Endocrinol,1991,129(5):2799-2801.
[1]Michael CR,Matlew JB,Virginia CP. Modulation of apoptosis by the widelydistributed bcl-2homology bax [J]. Nature,1995,374:736-738.
[2]Brey E M, Lalani Z, Johnston C, et al. Automated selection of DAB-labeled tissuefor immunohistochemical quantification [J].J Histochem Cytochem,2003,51(5):575-584.
[3]Maximova O A,Taffs R E, Pomeroy K L, et al. Computerized morphometricanalysis of pathological prion protein deposition in scrapie-infected hamster brain [J]. JHistochem Cytochem,2006,54(1):97-107.
[4]王树党.图像分析系统中光度学参数灰度与光密度的应用[J].山西医科大学学报,2003,34(5):462.
[5]李涛,范好,刘芳.免疫组织化学图像光密度分析的标准化方法[J].解剖学杂志,2008,31(5):727-728.
[6]崔涛,刘莹,朱祖安,等. NF-kB、Bcl-2、Bax在胃不典型增生、胃癌中的表达及其意义[J].山东大学学报,2006,44(7):689-693.
[7]牛建昭.组织学与胚胎学[M].第1版.北京:人民卫生出版社,2002:141-143.
[1]邝安堃.某些助阳药对于大剂量皮质激素所致竭现象的影响[J].中华内科杂志,1963,(2):651.
[2]陈英华,欧阳轶强,孙琪,等.肾阳虚证动物模型规范化研究诊断指标选择的初步探讨[J].中国中医基础医学杂志,2003,9(10):26-34.
[3]李广曦.肾阳虚证动物模型的造模方法及其相关指标回顾[J].中国中医基础医学杂志,2000,6(4):46-55.
[4]陈英华,孙琪,欧阳扶强,等.肾阳虚证动物模型造模方法综述[J].中国医药学报，2003,18(6):370-372.
[5]张伟荣.肾阳虚证动物实验研究简述及展望[J].中医药学刊,2006,24(10):1809-1811.
[6]董兴刚,张庆怡,陈以平.肾阳虚证的研究进展[J].中国中西医结合肾病杂志，2003,4(5):299-210.
[7]卢文丽,方肇勤.阳虚证动物模型的造模方法与评析[J].上海中医药大学学报,2004,18(4):45-49.
[8]徐敏,欧明,丘和明,等.甲状腺机能低下阳虚兔模型的造型研究[J].中医杂志,1990,(6):45.
[9]董兴刚,徐建国.肾切除加阿霉素诱导“肾阳虚”动物模型的研制[J].中国医药学报,2002,17(2):84-85.
[10]陈小野,邹世洁.大鼠肾上腺次全切除肾阳虚模型胃、舌病理观察[J].中国中医基础医学杂志,1999,5(11):20-23.
[11]邹世洁,陈小野,艾景录,等.大鼠肾上腺切除肾阳虚模型舌病理形态观察[J].四川中医,1997,15(11):5-6.
[12]陈小野,邹世洁,王震,等.大鼠肾上腺切除肾阳虚模型胃粘膜病理形态观察[J].天津中医,1997,14(6):267-269.
[13]岳广平,陈琼,戴宁.补肾生精汤对肾阳虚睾丸功能损害大鼠模型作用的实验研究[J].中国中西医结合杂志,1997,17(5):289-291.
[14]马威,薛莎,吴文莉,等.用血液微量元素观察右归丸治疗肾阳虚证的实验研究[J].微量元素与健康研究,1999,16(2):39-42.
[15]沈自尹,张丽丽,查良伦.肾阳虚病人的垂体—肾上腺系统的改变[J].上海中医药杂志,1979,(2):34-37.
[16]郭文峰,陈蔚文.脾虚证与营养物质吸收[J].新中医,2006,38(8):8-11.
[17]林传权,王桂香,李茹柳,等.慢性浅表性胃炎脾虚证消化吸收障碍亚型的临床分析[J].广州中医药大学学报,2008,25(6):488-491.
[18]郑小伟,王颖,宋红.补中益气汤对脾气虚证大鼠血清胃泌素影响的实验研究[J].中华中医药杂志(原中国医药学报),2006,21(7):393-395.
[19]郑小伟,宋红,包素珍,王颖,等.实验性脾气虚大鼠胃泌素的基因表达及四君子汤的干预作用[J].中华中医药杂志(原中国医药学报),2008,23(3):264-266.
[20]胡久略,贺又舜.参苓白术散对脾气虚证大鼠血清胃泌素影响的实验研究[J].时珍国医国药,2010,21(12):3346-3347.
[21]聂建华,欧阳文娟,阮时宝,等.土人参根健脾益气功效及其作用机制的实验研究[J].中国中医药科技,2009,16(3):200-201.
[22]张仲林,臧志和,钟玲,等.六君子汤对脾虚证大鼠胃肠激素影响的实验研究[J].中成药,2010,32(4):659-661.
[23]蔡华芳,蒋幼芳.小儿泄泻停冲剂的止泻及改善肠道吸收作用[J].儿科药学杂志,2005,11(6):3-4.
[24]朱曙东,李彬裴,乔樵.云母粉对脾虚泄泻证大鼠淀粉酶、木糖的影响[J].浙江中医学院学报,2002,26(6):46-7.
[25]田佳鑫,赖庆坤,马增春,等.腹泻康对脾虚泄泻证大鼠免疫和消化吸收功能的影响[J].中国临床康复,2006,10(47):92-94.
[26]王富春,逄紫千.针灸天枢穴对脾虚泄泻大鼠肠道功能影响的实验研究[J].长春中医学院学报,2005,21(1):52-54.
[27]徐丹,范颖.参附汤方源考证及其配伍内涵探析[J].中华中医药学刊,2010,28(5):1062-1063.
[28]国家药典委员会.中华人民共和国药典(2010年版1部)[M].第1版.北京:中国医药科技出版社,2010:356-358.
[29]王鸿燕.人参药对功用探析[J].中华中医药学刊,2009,27(3):649-651.
[30]国家药典委员会.中华人民共和国药典(2010年版1部)[M].第1版.北京:中国医药科技出版社,2010:10-11.
[31]秦华珍.人参还应归肾经[J].广西中医学院学报,2001,4(2):1-2.
[32]李德兴,王幼林,高长忠,等.中国乌头及附子对垂体-肾上腺皮质系统作用的研究[J].药学学报,1966,13(2):101-105.
[33]袁昌齐.天然药物资源开发利用[M].第1版.南京:江苏科学技术出版社,2000:4-5.
[34]刘伯顺,程秀娟.人参延缓衰老的研究[J].老年学杂志,1990,10(30)：190-192.
[1]金敬善,王广才,张绳组,等.血清中胃泌素水平与脾虚证的关系[J].中西医结合杂志.1982,2(1):25-26.
[2]姚永莉,宋于刚,张万岱.脾虚证与体液胃泌素及生长抑素关系的实验研究[J].中国中西医结合脾胃杂志,2000,8(6):333-335.
[3]郑小伟,王颖,宋红.三种脾气虚证模型大鼠血清胃泌素及胃窦G细胞的比较研究[J].中华中医药杂志,2006,21(6):338-340.
[4]张声生,王丽华,李颖辰,等.脾虚、脾虚痰湿证胃粘膜胃泌素和降钙素基因相关肽调节的研究[J].中国中医药信息杂志,2001,8(12):43-44.
[5]王丽华,张声生,李颖辰,等.脾虚、脾虚痰湿证血浆及胃粘膜胃肠激素水平的初步研究[J].北京中医杂志,2002,21(6):369-371.
[6]张声生,金敬善,赵荣莱,等.慢性胃炎和消化性溃疡患者血与尿前列腺素E2及前列腺素F2α水平的研究[J].中国中西医结合杂志,1992,12⑼:535-539.
[7]毛海燕.肝郁证大鼠肝线粒体膜流动性血浆胃动素及血清胃泌素的变化及意义[J].福建中医药,2003,34(3):42-43.
[8]程方平,刘松林,李家庚,等.温病湿热证大鼠模型胃动素胃泌素变化的对比研究[J].中华中医药学刊,2008,26(6):1350-1351.
[9]程方平,李家庚,刘松林,等.甘露消毒丹对湿热证大鼠模型的干预[J].中医药学报,2008,36(6):36-38.
[10]黄穗平,林燕钊.功能性消化不良不同分型血浆胃动素与胃泌素水平的对照研究[J].中医药学刊,2001,19(4):618-619.
[11]柴可夫,吴滇.功能性消化不良辨证分型的胃泌素、胃动素改变[J].中国中医基础医学杂志,1997,3(1):43-44.
[12]朱曙东.功能性消化不良中医证型与胃泌素胃动素关系的探讨[J].中西医杂志,1994,4(l):4-5.
[13]何美蓉,宋于刚,智发朝,等.功能性消化不良患者胃动力障碍与胃肠激素及G、D细胞的关系[J].中华消化杂志,2004,24(1):56-7.
[14]张建强,康美清.慢性萎缩性胃炎粘膜中胃泌素、生长抑素与脾胃虚弱证的相关性研究[J].中国中医急症,2010,19(5):786-788.
[15]邹科文,李其信,区显维,等.CSG中医辨证分型与血浆MTL、GAS、SS水平关系的临床研究[J].中国现代医生,2008,46(12):1-2,62.
[16]刘卫红,李萍,张蕾,等.慢性浅表性胃炎中医证型与胃粘膜分泌功能的相关性分析[J].北京中医,2006,25(12):709-711.
[17]蒋祖铭,邵文全.慢性胃炎的中医证型与胃肠激素关系[J].吉林中医药,2003,23(8):10-11.
[18]狄群英,倪宗珈,李兰芬,等.38例慢性胃部疾患病人血浆环核苷酸及血清胃泌素测定结果分析[J].云南中医中药杂志,1997,18(6):20-21.
[19]陆敏,赵卡冰,沈洪,等.幽门螺杆菌相关性慢性胃炎中医辨证分型与胃泌素的关系[J].中国中西医结合消化杂志,2004,12(1):21-22.
[20]马艳君,黄岑汉.慢性胃炎中医辨证分型与血清胃泌素和胃动素表达相关性研究[J].现代中西医结合杂志,2007,16(33):4914-4916.
[21]陈朝元,王岩,刘臣璋,等.消化性溃疡中医证型与血浆促胃泌素及生长抑素相关性研究[J].中国中西医结合杂志,2005,13(1):20-22.
[22]何应伟.176例慢性乙型肝炎中医辩证分型与血装胃泌素和甘胆酸含量变化的介析[J].中国中医药科技,1998,(5)3:141.
[23]崔丽安,张俊富,邵凤珍.慢性肝炎中医分型与客观化指标关系的探讨[J].山东中医杂志,2006,(25)1:23-26.
[24]杨敏春,许雅萍,张召才,等.不同中医证型脓毒症血小板和胃泌素的变化[J].浙江中西医结合杂志,2008,18(6):338-339.
[25]王少根,徐惠琴,王立基.严重烫伤早期血胃泌素、胃动素及β-内啡呔的测定和意义[J].中国病理生理杂志,2006,22(8):1653-1664
[26]李飞艳,邱赛红,尹健康,等.常用苦寒药对大鼠胃肠激素影响的研究[J].湖南中医药大学学报,2007,27(1):9-11.
[27]朱方石,王良静,姚健敏等.云母单体颗粒对萎缩性胃炎大鼠胃泌素、生长抑素及胃窦粘膜G,D细胞的影响[J].中国中药杂志,2004,29(6):554-558.
[28]王昕,滕静如,晋志高,等.西洋参汤对脾虚证大鼠血清中胃泌素、皮质醇含量的影响[J].中国中医基础医学杂志,2007,13(2):128-129.
[29]肖小河,赵艳玲,袁海龙,等.姜黄对胃泌素受体影响的研究[J].中药材,2002,25(3):184-185.
[30]李志,徐州,段国勋,等.香砂六君颗粒对脾虚大鼠胃肠动力和胃肠激素的影响[J].泸州医学院学报,2006,29(2):100-101.
[31]程艳波,张玉梅.锡类散对胃泌素及前列腺素E2分泌的影响[J].现代中西医结合杂志,2004,13(3):320.
[32]付丽丽,毛绍芳,王星田,等.舒肝解郁汤对慢性胃窦炎患者血清胃泌素及相关症状的影响[J].中国中西医结合消化杂志,2002,10(4):215-216.
[33]肖文,刘静,刘丽英.健脾温肾汤对糖尿病腹泻患者血清胃泌素、血浆胃动素及生长抑素的影响[J].中国中西医结合杂志,2002,22(8):587-589.
[34]张声生,陶琳,周斌,等.健脾理气消胀方治疗动力障碍型功能性消化不良的临床观察[J].中国中医药信息杂志,2000,7(12):53.
[35]王翼洲,张琳,高健.健脾调理汤治疗功能性消化不良的疗效观察[J].安徽医药,2004,8(5):337-339.
[36]刘松林,梅国强,赵映前,等.疏肝和胃汤对功能性消化不良大鼠血胃动素和胃泌素的影响[J].中国中西医结合消化杂志,2004,12(4):198-200.
[37]李洪梅,周爱香,李小芹,等.胃疡宁凝胶对大鼠乙酸烧灼型胃溃疡影响及其机理探讨[J].中国实验方剂学杂志,2005,11(1)：55-57.
[38]吕冠华,包永欣,劳绍贤.内外湿热因素对大鼠胃肠粘膜胃泌素、胃动素水平影响的研究[J].中国中医药科技2008,15(2):86-87.
[39]张声生,汪红兵,陶琳,等.健脾化湿法治疗腹泻型肠易激综合征及其对胃肠激素影响的研究[J].中国医药学报,2004,19(8):479-481.
[40]曾江涛,吴红苗,胡珂.辛开苦降法治疗寒热错杂型功能性消化不良的临床研究[J].新中医,2010,42(11):21-22.
[41]石光清,丁纯志,肖明.针刺足三里穴对血清胃泌素含量的影响[J].中国中西医结合杂志,2000,20(1):49.
[42]常小荣,严洁,林亚平,等.针刺足阳明经穴对健康人血浆胃动素、胃泌素含量的影响[J].中国中医药科技,2001,8(1):5.
[43]温庆祥,古颖.脾虚证手法治疗前后胃动素和胃泌素的变化[J].标记免疫分析与临床,2006,13(2):117-118.
[44]张锐,王联庆,李雨升,等.捏脊疗法对脾虚家兔血浆胃泌素的影响[J].按摩与导引,2004,20(5):5-7.
[45]周国平,严洁,常小荣,等.电针足三里对胃粘膜损伤家兔前列腺素E2、胃泌素水平的影响[J].中国中医药科技,2004,11(1):1-2.
[46]郭永明,梁宪如,邱桐,等.不同针刺手法对醋酸型胃溃疡大鼠溃疡指数及血清胃泌素水平的影响[J].天津中医学院学报,2001,20(4):27-28.
[47]朱元根,贲卉,王昕,等.耳-体穴电针对大鼠十二指肠组织中胃泌含量的影响及其促进胆汁分泌的作用[J].针刺研究,2000,25(4):272-275.
[48]吴学飞,郭永明,郑俊江,刘喆,等.温通针法对应激性胃粘膜损伤的保护作用[J].上海针灸杂志,2001,20(4):40-41.
[49]韩儒启.针灸对胃炎治疗作用及机理初探[J].陕西中医,2001,22(10)：620-621.
[2]吴敦序.中医基础理论[M].第1版.上海:上海科学技术出版社,2000:59.
[3]易青.再论“肾主水液”之内涵[J].湖北中医学院学报,2006,8(3):40-41.
[4]麻丽萍,易杰.论中医五脏生血[J].辽宁中医药大学学报,2009,11(8):5-6.
[5]吴志奎.肾生髓、髓生血理论与治疗地中海贫血的临床实践[J].中医杂志,2008,49(2):170-171.
[6]林殷,鲁兆麟.“补火生土”考辨[J].北京中医药大学学报,2003,26(3):11-13.
[7]蔡茂玉.脾主运化浅议[J].云南中医中药杂志,2004,25(3):58-59.
[1]戴芹,王怡,曲晓璐.温肾化痰方对早中期脾肾阳虚型慢性肾功能不全患者肾功能及血脂的影响[J].中国中西医结合杂志,2012,32(2):188-190.
[2]任可,马玲玲,刘美奇,等.血液透析后脾肾阳虚型患者血氧饱和度变化的临床观察[J].北京中医药大学学报（中医临床版）,2012,19(1):31-32.
[3]王晓英,苗得雨,裴妙荣.复合式造大鼠脾肾阳虚模型[J].山西中医,2012,28(7):43-46.
[4]林传权,王桂香,李茹柳,等.慢性浅表性胃炎脾虚证消化吸收障碍亚型的临床分析[J].广州中医药大学学报,2008,25(6):488-491.
[5]蔡茂玉.脾主运化浅议[J].云南中医中药杂志,2004,25(3):57-58.
[1]陈英华,欧阳轶强,孙琪,等.肾阳虚证动物模型规范化研究诊断指标选择的初步探讨[J].中国中医基础医学杂志,2003,9(10):26-34.
[2]李广曦.肾阳虚证动物模型的造模方法及其相关指标回顾[J].中国中医基础医学杂志,2000,6(4):46-55.
[3]陈小野,邹世洁,王震,等.大鼠肾上腺次全切除肾阳虚模型胃、舌病理观察[J].中国中医基础医学杂志,1999,5(11):20-23.
[4]苗明三.实验动物和动物实验技术[M].第1版.北京:中国中医药出版社,2003:242.
[5]贺石林,王键,王净净.中医科研设计与统计学[M].第1版.长沙:湖南科学技术出版社,2005:48-49.
[6]沈自尹,王文健.中医虚证辨证参考标准[J].中西医结合杂志，1986,6(10):598.
[1]邝安堃.中西医结合研究丛书—虚证研究[M].第1版.上海:上海科学技术出版社,1991:118.
[2]呼延和,张君.开郁健脾法对脾虚大鼠血清胃泌素、尿D-木糖影响的实验研究[J].中医儿科杂志,2010,6(3):12-16.
[3]李晓霞.脾气虚与Na+-K+-ATP酶活力观察[J].中国医学报,1996,11(2):83.
[4]Buts JP，Morin CL，Roy CC，et al. One-hour blood xylose test: a reliable index ofsmall bowel function [J].The Journal of Pediatries,1978,90(5):729-733.
[5]Yamamoto O, Matsunaga Y, Shiba Y,et al. Inhibition of motilin-induced phase Ⅲcontraction by pentagastrin in Heidenhain pouch dogs [J]. J Pharmacol ExpTher,1994,271:1471-1476.
[6]郑小伟,王颖,宋红.补中益气汤对脾气虚证大鼠血清胃泌素影响的实验研究[J].中华中医药杂志(原中国医药学报),2006,21(7):393-394.
[7]郑小伟,宋红,包素珍,等.实验性脾气虚大鼠胃泌素的基因表达及四君子汤的干预作用[J].中华中医药杂志(原中国医药学报),2008,23(3):264-266.
[1]王满贵,宋志宙.HE染色切片的技术操作规范[J].大同医学专科学校学报2004,(2):21.
[2]廉洁,张翠香,金明顺.HE染色的组织切片褪色的原因分析[J].齐齐哈尔医学院学报,2001,22(12):1432.
[3]蔡晓雯,沈武成.石蜡切片HE染色的体会[J].医药论坛杂志,2011,32(1):167-168.
[1]夏朝霞,何向蕾.原位细胞凋亡技术TUNEL法的改进[J].现代实用医学,2010,22(6):642-643.
[2]郭晓红,刘立新.Annexin V/PI流式细胞分析法和TUNEL法检测肝细胞凋亡的对比研究[J].山西医科大学学报[J].2008,39(5):476-479.
[3]李彦群,于凤玲,陈文雪.凋亡细胞TUNEL技术经验与改进[J].中国误诊学杂志,2001(1卷第10):1582-1583.
[4]董小莉,谭宁,符永恒,等.TUNEL法原位检测心肌细胞凋亡的改进.中国病理生理杂志,2010,26(5):1038-1040.
[5]张鸣号,彭亮,曹军.显微图像分析法与人工计数法在免疫组化结果判读中的应用[J].宁夏医科大学学报,2009,4(31):261-262.
[6]曹雪涛.免疫学技术及其应用[M].第1版.北京:科学出版社,2010:626.
[7]王伯沄,王文勇.免疫组织化学的新进展[J].中华医学会病理学分会学术年会论文汇编,2009:181.
[8]王秋生,马世文.免疫组化技术的应用[J].安庆医学,2004,25(1):37-38.
[9]许力,吴珊,杨清海,等.免疫组化技术(一)[J].诊断病理学杂志,2009,16,(2):158.
[10]石善溶.基于抗原修复技术的定量免疫组化:从实验、假说到研究计划[J].临床与实病理学杂志,2011,27(12):1271-1275.
[11]陈勤,周君,胡冠宇.石蜡切片免疫组化过程中存在的问题及解决方法[J].安徽大学学报(自然科学版),2009,33(2):85-88.
[12]高美钦,袁芳平,黄爱民,等.弹力纤维染色与免疫组织化学双重染色技术[J].中华病理学杂志,2003,32(2):176-177.
[13]应李雄,丁伟.DNA染色与免疫组织化学双重染色技术应用[J].中国组织化学与细胞化学杂志,2003,12(2):226-227.
[14]肖立,陆孝禹.CK34BE12和PSA顺序免疫组化双染色在前列腺病变中的表达[J].老年医学与保健,2003,9(1):14-18.
[15]陈罡,罗殿中,韦康来,等.双重免度组化染色法的操作体会[J].实用医技杂志,2004,11(5)下半月版:721-722.
[16]黄莺,王康敏,康安静,等.bFGF与Ki-67双重免疫组化染色法[J].诊断病理学杂志,2002,9(3):187.
[17]弓娟琴,陈志强,李文忠,等.Fas介导的凋亡与Caspase家族[J].国外医学:肿瘤学分册,2001,27(5):279-280．
[18]Stepien A, lzdebska M, Grzanka A. The types of cell death [J]. Postepy Hig MedDosw (Online),2007,61:420-428．
[19]Cryns V, Yuan J．Protease to die for [J]. Genes Dev,1998,12(11):1551-1570．
[20]赵霞,潘华峰,鞠晓云,等.胃痞消对CAG脾虚大鼠胃粘膜上皮细胞凋亡及调控蛋白Caspase-3影响的研究[J].中国中医基础医学杂志,2007,13(1):42-43.
[1]牛建昭.组织学与胚胎学[M].第1版.北京:人民卫生出版社,2002:141-143.
[2] Desoignie R,Sellin J. Propionate-initialed changes in intra-cellular pH in rabbitcoloncytes.Gastroenterology,1994,107(2):347-356.
[3]王辉,高颖.应用流式细胞仪的常见问题及解决对策[J].现代仪器,2012,18(1):64-65.
[4]胡凡.流式细胞仪在医学检验中的应用[J].中国保健营养,2012,(5):1139-1140.
[5] Brittan M, Wright NA. Stem cell in gastrointestinal structure and neoplasticdevelopment[J]. Gut,2004,53(6):899-910.
[6] Zhang C, Sheng ZY, Hu S, et al.The influence of apoptosis of mucosal epithelialcells on intestinal barrier integrity after scald in rats[J].Burns,2002,28(8):731-737.
[1]牛建昭.组织学与胚胎学[M].第1版.北京:人民卫生出版社,2002:141-143.
[2]冯新玲.恐伤孕鼠对其仔鼠脑发育的影响及其机理研究[D].湖北中医学院博士学位论文,2006:24.
[3] Otsuki Y. Various methods of apoptosis detection [J]. Acta HistochemCytochem,2000,33(4):235-241.
[4]Kalimichenko SG, Matveeva NY. Morphological characteristics of apoptosis and itssignificance in neurogenesis [J].Neurosci Behav Physiol,2008,38(4):333-344.
[5]Patel T, Gores JG. Apoptosis and hepatobiliary disease[J].Hepatol,1995,21(6):1725-1741.
[6]顾瑜蓉,李华伟,张重华,等.嗅球摘除后嗅觉神经元凋亡的实验研究[J].中华耳鼻咽喉头颈外科杂志,2006,41(4):297-300.
[7]沈强,俞彰,法京.电镜下几种凋亡细胞的形态特征[J].复旦学报(医学版),2010,37(3):322-325.
[8]Tilly JL, Kowalski KI, Johnson AL,et al. Involvement of apoptosis in ovarianfollicular atresia and postovulatory regression[J].Endocrinol,1991,129(5):2799-2801.
[1]Michael CR,Matlew JB,Virginia CP. Modulation of apoptosis by the widelydistributed bcl-2homology bax [J]. Nature,1995,374:736-738.
[2]Brey E M, Lalani Z, Johnston C, et al. Automated selection of DAB-labeled tissuefor immunohistochemical quantification [J].J Histochem Cytochem,2003,51(5):575-584.
[3]Maximova O A,Taffs R E, Pomeroy K L, et al. Computerized morphometricanalysis of pathological prion protein deposition in scrapie-infected hamster brain [J]. JHistochem Cytochem,2006,54(1):97-107.
[4]王树党.图像分析系统中光度学参数灰度与光密度的应用[J].山西医科大学学报,2003,34(5):462.
[5]李涛,范好,刘芳.免疫组织化学图像光密度分析的标准化方法[J].解剖学杂志,2008,31(5):727-728.
[6]崔涛,刘莹,朱祖安,等. NF-kB、Bcl-2、Bax在胃不典型增生、胃癌中的表达及其意义[J].山东大学学报,2006,44(7):689-693.
[7]牛建昭.组织学与胚胎学[M].第1版.北京:人民卫生出版社,2002:141-143.
[1]邝安堃.某些助阳药对于大剂量皮质激素所致竭现象的影响[J].中华内科杂志,1963,(2):651.
[2]陈英华,欧阳轶强,孙琪,等.肾阳虚证动物模型规范化研究诊断指标选择的初步探讨[J].中国中医基础医学杂志,2003,9(10):26-34.
[3]李广曦.肾阳虚证动物模型的造模方法及其相关指标回顾[J].中国中医基础医学杂志,2000,6(4):46-55.
[4]陈英华,孙琪,欧阳扶强,等.肾阳虚证动物模型造模方法综述[J].中国医药学报，2003,18(6):370-372.
[5]张伟荣.肾阳虚证动物实验研究简述及展望[J].中医药学刊,2006,24(10):1809-1811.
[6]董兴刚,张庆怡,陈以平.肾阳虚证的研究进展[J].中国中西医结合肾病杂志，2003,4(5):299-210.
[7]卢文丽,方肇勤.阳虚证动物模型的造模方法与评析[J].上海中医药大学学报,2004,18(4):45-49.
[8]徐敏,欧明,丘和明,等.甲状腺机能低下阳虚兔模型的造型研究[J].中医杂志,1990,(6):45.
[9]董兴刚,徐建国.肾切除加阿霉素诱导“肾阳虚”动物模型的研制[J].中国医药学报,2002,17(2):84-85.
[10]陈小野,邹世洁.大鼠肾上腺次全切除肾阳虚模型胃、舌病理观察[J].中国中医基础医学杂志,1999,5(11):20-23.
[11]邹世洁,陈小野,艾景录,等.大鼠肾上腺切除肾阳虚模型舌病理形态观察[J].四川中医,1997,15(11):5-6.
[12]陈小野,邹世洁,王震,等.大鼠肾上腺切除肾阳虚模型胃粘膜病理形态观察[J].天津中医,1997,14(6):267-269.
[13]岳广平,陈琼,戴宁.补肾生精汤对肾阳虚睾丸功能损害大鼠模型作用的实验研究[J].中国中西医结合杂志,1997,17(5):289-291.
[14]马威,薛莎,吴文莉,等.用血液微量元素观察右归丸治疗肾阳虚证的实验研究[J].微量元素与健康研究,1999,16(2):39-42.
[15]沈自尹,张丽丽,查良伦.肾阳虚病人的垂体—肾上腺系统的改变[J].上海中医药杂志,1979,(2):34-37.
[16]郭文峰,陈蔚文.脾虚证与营养物质吸收[J].新中医,2006,38(8):8-11.
[17]林传权,王桂香,李茹柳,等.慢性浅表性胃炎脾虚证消化吸收障碍亚型的临床分析[J].广州中医药大学学报,2008,25(6):488-491.
[18]郑小伟,王颖,宋红.补中益气汤对脾气虚证大鼠血清胃泌素影响的实验研究[J].中华中医药杂志(原中国医药学报),2006,21(7):393-395.
[19]郑小伟,宋红,包素珍,王颖,等.实验性脾气虚大鼠胃泌素的基因表达及四君子汤的干预作用[J].中华中医药杂志(原中国医药学报),2008,23(3):264-266.
[20]胡久略,贺又舜.参苓白术散对脾气虚证大鼠血清胃泌素影响的实验研究[J].时珍国医国药,2010,21(12):3346-3347.
[21]聂建华,欧阳文娟,阮时宝,等.土人参根健脾益气功效及其作用机制的实验研究[J].中国中医药科技,2009,16(3):200-201.
[22]张仲林,臧志和,钟玲,等.六君子汤对脾虚证大鼠胃肠激素影响的实验研究[J].中成药,2010,32(4):659-661.
[23]蔡华芳,蒋幼芳.小儿泄泻停冲剂的止泻及改善肠道吸收作用[J].儿科药学杂志,2005,11(6):3-4.
[24]朱曙东,李彬裴,乔樵.云母粉对脾虚泄泻证大鼠淀粉酶、木糖的影响[J].浙江中医学院学报,2002,26(6):46-7.
[25]田佳鑫,赖庆坤,马增春,等.腹泻康对脾虚泄泻证大鼠免疫和消化吸收功能的影响[J].中国临床康复,2006,10(47):92-94.
[26]王富春,逄紫千.针灸天枢穴对脾虚泄泻大鼠肠道功能影响的实验研究[J].长春中医学院学报,2005,21(1):52-54.
[27]徐丹,范颖.参附汤方源考证及其配伍内涵探析[J].中华中医药学刊,2010,28(5):1062-1063.
[28]国家药典委员会.中华人民共和国药典(2010年版1部)[M].第1版.北京:中国医药科技出版社,2010:356-358.
[29]王鸿燕.人参药对功用探析[J].中华中医药学刊,2009,27(3):649-651.
[30]国家药典委员会.中华人民共和国药典(2010年版1部)[M].第1版.北京:中国医药科技出版社,2010:10-11.
[31]秦华珍.人参还应归肾经[J].广西中医学院学报,2001,4(2):1-2.
[32]李德兴,王幼林,高长忠,等.中国乌头及附子对垂体-肾上腺皮质系统作用的研究[J].药学学报,1966,13(2):101-105.
[33]袁昌齐.天然药物资源开发利用[M].第1版.南京:江苏科学技术出版社,2000:4-5.
[34]刘伯顺,程秀娟.人参延缓衰老的研究[J].老年学杂志,1990,10(30)：190-192.
[1]金敬善,王广才,张绳组,等.血清中胃泌素水平与脾虚证的关系[J].中西医结合杂志.1982,2(1):25-26.
[2]姚永莉,宋于刚,张万岱.脾虚证与体液胃泌素及生长抑素关系的实验研究[J].中国中西医结合脾胃杂志,2000,8(6):333-335.
[3]郑小伟,王颖,宋红.三种脾气虚证模型大鼠血清胃泌素及胃窦G细胞的比较研究[J].中华中医药杂志,2006,21(6):338-340.
[4]张声生,王丽华,李颖辰,等.脾虚、脾虚痰湿证胃粘膜胃泌素和降钙素基因相关肽调节的研究[J].中国中医药信息杂志,2001,8(12):43-44.
[5]王丽华,张声生,李颖辰,等.脾虚、脾虚痰湿证血浆及胃粘膜胃肠激素水平的初步研究[J].北京中医杂志,2002,21(6):369-371.
[6]张声生,金敬善,赵荣莱,等.慢性胃炎和消化性溃疡患者血与尿前列腺素E2及前列腺素F2α水平的研究[J].中国中西医结合杂志,1992,12⑼:535-539.
[7]毛海燕.肝郁证大鼠肝线粒体膜流动性血浆胃动素及血清胃泌素的变化及意义[J].福建中医药,2003,34(3):42-43.
[8]程方平,刘松林,李家庚,等.温病湿热证大鼠模型胃动素胃泌素变化的对比研究[J].中华中医药学刊,2008,26(6):1350-1351.
[9]程方平,李家庚,刘松林,等.甘露消毒丹对湿热证大鼠模型的干预[J].中医药学报,2008,36(6):36-38.
[10]黄穗平,林燕钊.功能性消化不良不同分型血浆胃动素与胃泌素水平的对照研究[J].中医药学刊,2001,19(4):618-619.
[11]柴可夫,吴滇.功能性消化不良辨证分型的胃泌素、胃动素改变[J].中国中医基础医学杂志,1997,3(1):43-44.
[12]朱曙东.功能性消化不良中医证型与胃泌素胃动素关系的探讨[J].中西医杂志,1994,4(l):4-5.
[13]何美蓉,宋于刚,智发朝,等.功能性消化不良患者胃动力障碍与胃肠激素及G、D细胞的关系[J].中华消化杂志,2004,24(1):56-7.
[14]张建强,康美清.慢性萎缩性胃炎粘膜中胃泌素、生长抑素与脾胃虚弱证的相关性研究[J].中国中医急症,2010,19(5):786-788.
[15]邹科文,李其信,区显维,等.CSG中医辨证分型与血浆MTL、GAS、SS水平关系的临床研究[J].中国现代医生,2008,46(12):1-2,62.
[16]刘卫红,李萍,张蕾,等.慢性浅表性胃炎中医证型与胃粘膜分泌功能的相关性分析[J].北京中医,2006,25(12):709-711.
[17]蒋祖铭,邵文全.慢性胃炎的中医证型与胃肠激素关系[J].吉林中医药,2003,23(8):10-11.
[18]狄群英,倪宗珈,李兰芬,等.38例慢性胃部疾患病人血浆环核苷酸及血清胃泌素测定结果分析[J].云南中医中药杂志,1997,18(6):20-21.
[19]陆敏,赵卡冰,沈洪,等.幽门螺杆菌相关性慢性胃炎中医辨证分型与胃泌素的关系[J].中国中西医结合消化杂志,2004,12(1):21-22.
[20]马艳君,黄岑汉.慢性胃炎中医辨证分型与血清胃泌素和胃动素表达相关性研究[J].现代中西医结合杂志,2007,16(33):4914-4916.
[21]陈朝元,王岩,刘臣璋,等.消化性溃疡中医证型与血浆促胃泌素及生长抑素相关性研究[J].中国中西医结合杂志,2005,13(1):20-22.
[22]何应伟.176例慢性乙型肝炎中医辩证分型与血装胃泌素和甘胆酸含量变化的介析[J].中国中医药科技,1998,(5)3:141.
[23]崔丽安,张俊富,邵凤珍.慢性肝炎中医分型与客观化指标关系的探讨[J].山东中医杂志,2006,(25)1:23-26.
[24]杨敏春,许雅萍,张召才,等.不同中医证型脓毒症血小板和胃泌素的变化[J].浙江中西医结合杂志,2008,18(6):338-339.
[25]王少根,徐惠琴,王立基.严重烫伤早期血胃泌素、胃动素及β-内啡呔的测定和意义[J].中国病理生理杂志,2006,22(8):1653-1664
[26]李飞艳,邱赛红,尹健康,等.常用苦寒药对大鼠胃肠激素影响的研究[J].湖南中医药大学学报,2007,27(1):9-11.
[27]朱方石,王良静,姚健敏等.云母单体颗粒对萎缩性胃炎大鼠胃泌素、生长抑素及胃窦粘膜G,D细胞的影响[J].中国中药杂志,2004,29(6):554-558.
[28]王昕,滕静如,晋志高,等.西洋参汤对脾虚证大鼠血清中胃泌素、皮质醇含量的影响[J].中国中医基础医学杂志,2007,13(2):128-129.
[29]肖小河,赵艳玲,袁海龙,等.姜黄对胃泌素受体影响的研究[J].中药材,2002,25(3):184-185.
[30]李志,徐州,段国勋,等.香砂六君颗粒对脾虚大鼠胃肠动力和胃肠激素的影响[J].泸州医学院学报,2006,29(2):100-101.
[31]程艳波,张玉梅.锡类散对胃泌素及前列腺素E2分泌的影响[J].现代中西医结合杂志,2004,13(3):320.
[32]付丽丽,毛绍芳,王星田,等.舒肝解郁汤对慢性胃窦炎患者血清胃泌素及相关症状的影响[J].中国中西医结合消化杂志,2002,10(4):215-216.
[33]肖文,刘静,刘丽英.健脾温肾汤对糖尿病腹泻患者血清胃泌素、血浆胃动素及生长抑素的影响[J].中国中西医结合杂志,2002,22(8):587-589.
[34]张声生,陶琳,周斌,等.健脾理气消胀方治疗动力障碍型功能性消化不良的临床观察[J].中国中医药信息杂志,2000,7(12):53.
[35]王翼洲,张琳,高健.健脾调理汤治疗功能性消化不良的疗效观察[J].安徽医药,2004,8(5):337-339.
[36]刘松林,梅国强,赵映前,等.疏肝和胃汤对功能性消化不良大鼠血胃动素和胃泌素的影响[J].中国中西医结合消化杂志,2004,12(4):198-200.
[37]李洪梅,周爱香,李小芹,等.胃疡宁凝胶对大鼠乙酸烧灼型胃溃疡影响及其机理探讨[J].中国实验方剂学杂志,2005,11(1)：55-57.
[38]吕冠华,包永欣,劳绍贤.内外湿热因素对大鼠胃肠粘膜胃泌素、胃动素水平影响的研究[J].中国中医药科技2008,15(2):86-87.
[39]张声生,汪红兵,陶琳,等.健脾化湿法治疗腹泻型肠易激综合征及其对胃肠激素影响的研究[J].中国医药学报,2004,19(8):479-481.
[40]曾江涛,吴红苗,胡珂.辛开苦降法治疗寒热错杂型功能性消化不良的临床研究[J].新中医,2010,42(11):21-22.
[41]石光清,丁纯志,肖明.针刺足三里穴对血清胃泌素含量的影响[J].中国中西医结合杂志,2000,20(1):49.
[42]常小荣,严洁,林亚平,等.针刺足阳明经穴对健康人血浆胃动素、胃泌素含量的影响[J].中国中医药科技,2001,8(1):5.
[43]温庆祥,古颖.脾虚证手法治疗前后胃动素和胃泌素的变化[J].标记免疫分析与临床,2006,13(2):117-118.
[44]张锐,王联庆,李雨升,等.捏脊疗法对脾虚家兔血浆胃泌素的影响[J].按摩与导引,2004,20(5):5-7.
[45]周国平,严洁,常小荣,等.电针足三里对胃粘膜损伤家兔前列腺素E2、胃泌素水平的影响[J].中国中医药科技,2004,11(1):1-2.
[46]郭永明,梁宪如,邱桐,等.不同针刺手法对醋酸型胃溃疡大鼠溃疡指数及血清胃泌素水平的影响[J].天津中医学院学报,2001,20(4):27-28.
[47]朱元根,贲卉,王昕,等.耳-体穴电针对大鼠十二指肠组织中胃泌含量的影响及其促进胆汁分泌的作用[J].针刺研究,2000,25(4):272-275.
[48]吴学飞,郭永明,郑俊江,刘喆,等.温通针法对应激性胃粘膜损伤的保护作用[J].上海针灸杂志,2001,20(4):40-41.
[49]韩儒启.针灸对胃炎治疗作用及机理初探[J].陕西中医,2001,22(10)：620-621.