电针对原发性痛经模型大鼠镇痛效应及免疫机制的研究
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摘要
研究目的:
原发性痛经为妇科常见病之一,运用中医治疗效果显著。其发病机制复杂,可能与内分泌失调、子宫收缩异常、以及子宫缺血、缺氧有关。本文从针灸的角度探讨在痛经治疗方面之研究进展。近年来,针灸疗法在治疗原发性痛经方面的研究报导很多,因施术简单、疗效确切、副作用少而受到医学界普遍关注。希望借着此课题,在前人的研究及经验累积上,进一步探讨治疗原发性痛经有效、安全、可重复性的方法。
研究方法:
(一)文献研究
采用近20年的文献研究方法,针对现代医学对原发性痛经机理的认识及其治疗痛经常用方法综合阐述,并由中医对痛经的认识及针灸治疗痛经机理研究的观点切入,进行讨论与展望。
(二)实验研究
本研究以原发性痛经机体的反应及其调节能力为切入点,采用生物化学、放射免疫学等方法,从整体、系统、器官、分子等不同水平观察原发性痛经模型大鼠,并与西药组比较,以评价原发性痛经模型大鼠针刺的治疗作用。
1、实验动物与分组:清洁级健康雌性SD大鼠40只。随机分成空白对照组、模型组、电针组、西药组,每组各10只大鼠。
2、原发性痛经模型造模方法:连续10天给予给予股部皮下注射已烯雌酚,每天一次,第一天0.8mg/只,第2-9天0.4mg/只,第10天0.8mg/只;第11天腹腔注射缩宫素2U/只。
3、电针穴位处方:依据中医认为月经与冲任二脉关系最为密切,而痛经与肝脾肾三脏功能失调有关的理论,和导师学术思想指导,依调气行血,温经散寒,行气止痛的组方原则,制定穴位处方:三阴交_双,关元。
4、西药组:连续10天给予灌胃给药0.8ml/只(生理盐水配制,浓度为125mg/100ml)每天一次。
5、观察原发性痛经模型大鼠一般行为和扭体反应。
6、观察原发性痛经模型大鼠子宫内膜PGE_2、PGF_(2a)变化。
7、观察原发性痛经模型大鼠血清TNF-α、IL-6、IL-1变化。
8、观察原发性痛经模型大鼠脾脏NK细胞活性变化。
9、观察原发性痛经模型大鼠下丘脑组织中5-羟色胺含量的变化。
10、观察电针对原发性痛经模型大鼠上述改变的影响,以探讨电针治疗原发性痛经的作用与机制。
11、分析上述相关指标改变的意义及其指标之间的相互关系。
结果:
1、原发性痛经模型大鼠毛发色泽暗淡萎黄,动作反应及新陈代谢变缓,精神状态变差。经过电针治疗与服用芬必得皆可减缓模型大鼠基本体征的改变。
2、原发性痛经模型大鼠子宫收缩的扭体反应中,各组与模型组比较,反应次数明显较少;正常对造组的大鼠并不具有扭体反应,与西药组、电针治疗组比较具有差异性;西药组、电针治疗组之间并无显著性差异,提示大鼠造模成功,只要经过治疗不论是西药组或是电针组都能改善大鼠扭体反应。
3、原发性痛经模型大鼠子宫内膜PGF_(2a)含量正常对照组与电针组、西药组、模型组各组之间显着不同,由低而高排列为正常对照组、西药组、电针组、模型组;PGE_2含量各组之间也有显著差异,由低而高排列为模型组、正常对照组、电针组、西药组;PGF_(2a)/PGE_2的比值在模型组与正常对照组、电针组、西药组之间有显着差异,在正常对照组与电针组、西药组之间并无显著差异,电针组与西药组相比,有显着差异。提示依据疼痛的不同程度,PGF_(2a)含量也有所不同,含量愈高,疼痛愈明显,与之相反的是PGE_2含量愈低,疼痛愈明显。而疼痛的最后表现在于PGF_(2a)/PGE_2的比值,由结果得知各组疼痛程度与模型组有显著差异,但经过针刺或是服用芬必得后,疼痛程度降低,与正常对照组无异,但西药组的镇痛效果优于电针组。
4、原发性痛经模型大鼠血清中TNF-α、IL-6、IL-1含量、各组与模型组相比,浓度明显降低;西药组、电针组与正常对造组之间并无显著性差异。大鼠脾脏NK细胞活性各组与模型组相比,浓度明显升高;西药组、电针组与正常对造组之间并无显著性差异。原发性痛经模型大鼠下丘脑组织中5-羟色胺含量各组与模型组相比,含量有显着增高,提示造模后未经西药或是电针治疗的大鼠下丘脑组织中5-羟色胺(5-HT)含量与各组确实存在差异性。电针组与西药组与正常对照组大鼠下丘脑组织中5-羟色胺含量无明显变化;提示了电针可提高模型大鼠血清与脾脏中的免疫防御系统的功能,与服用芬必得的解痉镇痛作用无显著差异,经过电针或是西药治疗后,机体下丘脑组织中5-羟色胺含量可达到正常对照组的水平,不论是服用西药或是电针刺治疗原发性痛经都有一定的缓解疼痛效果。
5、观察原发性痛经机体的PGE_2、PGF_(2a)、TNF-α、IL-6、IL-1、TNF-α、IL-6、IL-1、5-HT的水平,利用中医治疗痛经的理论,提供了临床应用电针治疗痛经的依据。
结论:
1、经由扭体实验的观察说明了本次实验造模是成功的。
2、电针治疗可以改善原发性痛经大鼠模型PGE_2、PGF_(2a)、TNF-α、IL-6、IL-1、TNF-α、IL-6、IL-1、5-HT的水平,本研究提供了临床应用电针治疗痛经的依据,为后续的深入研究奠定了基础。
在针灸治疗痛经研究中,对针刺治疗原发性痛经的免疫机制进行了研究。从免疫学角度探讨PD的形成机制及针刺治疗原发性痛经的获效机理,进而研究针刺治疗该病的免疫机制,是本研究的创新之处。祖国医学在治疗痛经方面积累了非常丰富的经验,针灸用于治疗月经病的历史久远,操作简便、价廉、无毒副作用、适应症广、效果显著、具有独特优势,针灸对机体具有明显的良性调整作用,特别是针灸医学的整体调节和保健作用,对治疗痛经有着无可比拟的优势。
1.Reserch purpose
Dysmenorrhea is a common disease in the department of gynecology and can be treated effectively by acupuncture and moxibution.Its pathogenesis is very complicated and probably close to endocrine unbalance abnormal uterine contractility,uterine ischemia of hypoxia.The subject of this article is to explore the new development of TCM treatments on dysmenorrhea application. In recent years there have been many reports on the use of acupuncture in treating PD,because its applications are simple,effective,and with few side effects it has received widespread attention from the medical community. Hopefully by addressing this issue,we can further investigate treatment of PD by building on previously accumulated research and experience,to find a method of treatment that is safe,effective,and can be repeated.
2.Method
2.1 Literature Research
Adopt documents researching approach to explain synthetically the understanding of primary dysmenorrhea mechanism and commonly used methods of treating dysmenorrhea in accordance to symptoms,the TCM understanding of primary dysmenorrhea,and treating dysmenorrhea with acupuncture therapy and moxibustion,and to comment and look into the distance.
2.2 Experimental Research
This research regards the equilibrium ability of the PD organism as the breakthrough point;adopts methods include biochemistry,radioimmunology etc; observes and appraises,the effects of electro acupuncture therapy under different groups with individual levels which are a whole,a system,an organ, and a molecule and compares and contrasts it to the effects achieved by Western medication.
(1) The animals tested upon and grouped as of the following:40 SD female rats of clean class,were divided randomly into four groups,the blank group, the model group,the electric needle group,and the medication group,with each group containing 10 rats.
(2) Establishing the PD animal model:Make the model by injecting 0.8mg of diethylstilbestrol injection per rat(1~(st) and 10~(th) day),0.4mg per rat(from 2~(nd) day to 9~(th) day) into the abdominal cavity and 2U of oxytocin injection per rat on the 11~(th) day.
(3) Method of choosing electro acupuncture prescriptions:According to the traditional theory under TCM of the Ren meridian,Chong meridian, prescription composed of Guanyuan(RN4),Sanyyinjiao(SP6).
(4) Medication group:Make model by feeding 0.8ml of Ibuprofen sustained-release capsules per rat for 10 days in succession.
(5) Observe general behavior and amount and percentage of total found squirming.
(6) Observe PGE_2,PGF_(2a) contents of endometrium of the PD model rats under the different groups.
(7) Observe TNF-α,IL-6,IL-1 contents of serum of the PD model rats under the different groups.
(8) Observe NK cell activeness in the spleen of the PD model rats under the different groups.
(9) Observe 5-HT contents of the hypothalamic organization of the PD model rats under the different groups.
(10) Observe the change and influence of electro acupuncture on the above mentioned in order to probe into its function and mechanism of treating PD.
(11) Analyze the meaning of the change of the above mentioned and its relevant index and interaction between meaning and index in which the above mentioned' s relevant indexes change.
3.Result
(1) The PD model rats appeared with slow movement and reaction,dispirited spirit,weight increase getting slow,the hair color turned yellow and the gloss was bad.Electro acupuncture could improve this status.
(2) Not only electro acupuncture but also western medicine could improve body-stretching rate.
(3) To observe endometrium contents of PGE_2,PGF_(2a),the ratio of PGF_(2a)/PGE_2 of the PD model rats under the different groups,there exists an obvious difference.There is no obvious difference between the blank group,electro acupuncture group and medication group but electro acupuncture group and medication group.
(4) The electro acupuncture can improve the serum contents of TNF-α, IL-6,IL-1 and spleen NK cell activeness and hypothalamic organization contents of 5-HI under the PD model,enhance the function of immunity systems. The pain release effect is the same as western medicine and achieved the normal rats standard.
(5) To observe the response to the PD rats under different groups,use the TCM principles with acupuncture,which provide experimental data for clinical application and a basis for further study.
4.Conclusions
(1) The model was made successfully according to the observation of general behavior and amount and percentage of total found squirming.
(2) The electro acupuncture can improve the level of PGE_2,PGF_(2a),TNF-α, IL-6,IL-1,TNF-α,IL-6,IL-1 and 5-HT.This research provides experimental data for clinical application of treating PD by electro acupuncture and a basis for further study.
There is abundant experience on PD treatment under TCM.To use the acupuncture and moxibustion to treat PD is very special and strong.It is cheaper,easier,non-poison etc.
原发性痛经为妇科常见病之一,运用中医治疗效果显著。其发病机制复杂,可能与内分泌失调、子宫收缩异常、以及子宫缺血、缺氧有关。本文从针灸的角度探讨在痛经治疗方面之研究进展。近年来,针灸疗法在治疗原发性痛经方面的研究报导很多,因施术简单、疗效确切、副作用少而受到医学界普遍关注。希望借着此课题,在前人的研究及经验累积上,进一步探讨治疗原发性痛经有效、安全、可重复性的方法。
研究方法:
(一)文献研究
采用近20年的文献研究方法,针对现代医学对原发性痛经机理的认识及其治疗痛经常用方法综合阐述,并由中医对痛经的认识及针灸治疗痛经机理研究的观点切入,进行讨论与展望。
(二)实验研究
本研究以原发性痛经机体的反应及其调节能力为切入点,采用生物化学、放射免疫学等方法,从整体、系统、器官、分子等不同水平观察原发性痛经模型大鼠,并与西药组比较,以评价原发性痛经模型大鼠针刺的治疗作用。
1、实验动物与分组:清洁级健康雌性SD大鼠40只。随机分成空白对照组、模型组、电针组、西药组,每组各10只大鼠。
2、原发性痛经模型造模方法:连续10天给予给予股部皮下注射已烯雌酚,每天一次,第一天0.8mg/只,第2-9天0.4mg/只,第10天0.8mg/只;第11天腹腔注射缩宫素2U/只。
3、电针穴位处方:依据中医认为月经与冲任二脉关系最为密切,而痛经与肝脾肾三脏功能失调有关的理论,和导师学术思想指导,依调气行血,温经散寒,行气止痛的组方原则,制定穴位处方:三阴交_双,关元。
4、西药组:连续10天给予灌胃给药0.8ml/只(生理盐水配制,浓度为125mg/100ml)每天一次。
5、观察原发性痛经模型大鼠一般行为和扭体反应。
6、观察原发性痛经模型大鼠子宫内膜PGE_2、PGF_(2a)变化。
7、观察原发性痛经模型大鼠血清TNF-α、IL-6、IL-1变化。
8、观察原发性痛经模型大鼠脾脏NK细胞活性变化。
9、观察原发性痛经模型大鼠下丘脑组织中5-羟色胺含量的变化。
10、观察电针对原发性痛经模型大鼠上述改变的影响,以探讨电针治疗原发性痛经的作用与机制。
11、分析上述相关指标改变的意义及其指标之间的相互关系。
结果:
1、原发性痛经模型大鼠毛发色泽暗淡萎黄,动作反应及新陈代谢变缓,精神状态变差。经过电针治疗与服用芬必得皆可减缓模型大鼠基本体征的改变。
2、原发性痛经模型大鼠子宫收缩的扭体反应中,各组与模型组比较,反应次数明显较少;正常对造组的大鼠并不具有扭体反应,与西药组、电针治疗组比较具有差异性;西药组、电针治疗组之间并无显著性差异,提示大鼠造模成功,只要经过治疗不论是西药组或是电针组都能改善大鼠扭体反应。
3、原发性痛经模型大鼠子宫内膜PGF_(2a)含量正常对照组与电针组、西药组、模型组各组之间显着不同,由低而高排列为正常对照组、西药组、电针组、模型组;PGE_2含量各组之间也有显著差异,由低而高排列为模型组、正常对照组、电针组、西药组;PGF_(2a)/PGE_2的比值在模型组与正常对照组、电针组、西药组之间有显着差异,在正常对照组与电针组、西药组之间并无显著差异,电针组与西药组相比,有显着差异。提示依据疼痛的不同程度,PGF_(2a)含量也有所不同,含量愈高,疼痛愈明显,与之相反的是PGE_2含量愈低,疼痛愈明显。而疼痛的最后表现在于PGF_(2a)/PGE_2的比值,由结果得知各组疼痛程度与模型组有显著差异,但经过针刺或是服用芬必得后,疼痛程度降低,与正常对照组无异,但西药组的镇痛效果优于电针组。
4、原发性痛经模型大鼠血清中TNF-α、IL-6、IL-1含量、各组与模型组相比,浓度明显降低;西药组、电针组与正常对造组之间并无显著性差异。大鼠脾脏NK细胞活性各组与模型组相比,浓度明显升高;西药组、电针组与正常对造组之间并无显著性差异。原发性痛经模型大鼠下丘脑组织中5-羟色胺含量各组与模型组相比,含量有显着增高,提示造模后未经西药或是电针治疗的大鼠下丘脑组织中5-羟色胺(5-HT)含量与各组确实存在差异性。电针组与西药组与正常对照组大鼠下丘脑组织中5-羟色胺含量无明显变化;提示了电针可提高模型大鼠血清与脾脏中的免疫防御系统的功能,与服用芬必得的解痉镇痛作用无显著差异,经过电针或是西药治疗后,机体下丘脑组织中5-羟色胺含量可达到正常对照组的水平,不论是服用西药或是电针刺治疗原发性痛经都有一定的缓解疼痛效果。
5、观察原发性痛经机体的PGE_2、PGF_(2a)、TNF-α、IL-6、IL-1、TNF-α、IL-6、IL-1、5-HT的水平,利用中医治疗痛经的理论,提供了临床应用电针治疗痛经的依据。
结论:
1、经由扭体实验的观察说明了本次实验造模是成功的。
2、电针治疗可以改善原发性痛经大鼠模型PGE_2、PGF_(2a)、TNF-α、IL-6、IL-1、TNF-α、IL-6、IL-1、5-HT的水平,本研究提供了临床应用电针治疗痛经的依据,为后续的深入研究奠定了基础。
在针灸治疗痛经研究中,对针刺治疗原发性痛经的免疫机制进行了研究。从免疫学角度探讨PD的形成机制及针刺治疗原发性痛经的获效机理,进而研究针刺治疗该病的免疫机制,是本研究的创新之处。祖国医学在治疗痛经方面积累了非常丰富的经验,针灸用于治疗月经病的历史久远,操作简便、价廉、无毒副作用、适应症广、效果显著、具有独特优势,针灸对机体具有明显的良性调整作用,特别是针灸医学的整体调节和保健作用,对治疗痛经有着无可比拟的优势。
1.Reserch purpose
Dysmenorrhea is a common disease in the department of gynecology and can be treated effectively by acupuncture and moxibution.Its pathogenesis is very complicated and probably close to endocrine unbalance abnormal uterine contractility,uterine ischemia of hypoxia.The subject of this article is to explore the new development of TCM treatments on dysmenorrhea application. In recent years there have been many reports on the use of acupuncture in treating PD,because its applications are simple,effective,and with few side effects it has received widespread attention from the medical community. Hopefully by addressing this issue,we can further investigate treatment of PD by building on previously accumulated research and experience,to find a method of treatment that is safe,effective,and can be repeated.
2.Method
2.1 Literature Research
Adopt documents researching approach to explain synthetically the understanding of primary dysmenorrhea mechanism and commonly used methods of treating dysmenorrhea in accordance to symptoms,the TCM understanding of primary dysmenorrhea,and treating dysmenorrhea with acupuncture therapy and moxibustion,and to comment and look into the distance.
2.2 Experimental Research
This research regards the equilibrium ability of the PD organism as the breakthrough point;adopts methods include biochemistry,radioimmunology etc; observes and appraises,the effects of electro acupuncture therapy under different groups with individual levels which are a whole,a system,an organ, and a molecule and compares and contrasts it to the effects achieved by Western medication.
(1) The animals tested upon and grouped as of the following:40 SD female rats of clean class,were divided randomly into four groups,the blank group, the model group,the electric needle group,and the medication group,with each group containing 10 rats.
(2) Establishing the PD animal model:Make the model by injecting 0.8mg of diethylstilbestrol injection per rat(1~(st) and 10~(th) day),0.4mg per rat(from 2~(nd) day to 9~(th) day) into the abdominal cavity and 2U of oxytocin injection per rat on the 11~(th) day.
(3) Method of choosing electro acupuncture prescriptions:According to the traditional theory under TCM of the Ren meridian,Chong meridian, prescription composed of Guanyuan(RN4),Sanyyinjiao(SP6).
(4) Medication group:Make model by feeding 0.8ml of Ibuprofen sustained-release capsules per rat for 10 days in succession.
(5) Observe general behavior and amount and percentage of total found squirming.
(6) Observe PGE_2,PGF_(2a) contents of endometrium of the PD model rats under the different groups.
(7) Observe TNF-α,IL-6,IL-1 contents of serum of the PD model rats under the different groups.
(8) Observe NK cell activeness in the spleen of the PD model rats under the different groups.
(9) Observe 5-HT contents of the hypothalamic organization of the PD model rats under the different groups.
(10) Observe the change and influence of electro acupuncture on the above mentioned in order to probe into its function and mechanism of treating PD.
(11) Analyze the meaning of the change of the above mentioned and its relevant index and interaction between meaning and index in which the above mentioned' s relevant indexes change.
3.Result
(1) The PD model rats appeared with slow movement and reaction,dispirited spirit,weight increase getting slow,the hair color turned yellow and the gloss was bad.Electro acupuncture could improve this status.
(2) Not only electro acupuncture but also western medicine could improve body-stretching rate.
(3) To observe endometrium contents of PGE_2,PGF_(2a),the ratio of PGF_(2a)/PGE_2 of the PD model rats under the different groups,there exists an obvious difference.There is no obvious difference between the blank group,electro acupuncture group and medication group but electro acupuncture group and medication group.
(4) The electro acupuncture can improve the serum contents of TNF-α, IL-6,IL-1 and spleen NK cell activeness and hypothalamic organization contents of 5-HI under the PD model,enhance the function of immunity systems. The pain release effect is the same as western medicine and achieved the normal rats standard.
(5) To observe the response to the PD rats under different groups,use the TCM principles with acupuncture,which provide experimental data for clinical application and a basis for further study.
4.Conclusions
(1) The model was made successfully according to the observation of general behavior and amount and percentage of total found squirming.
(2) The electro acupuncture can improve the level of PGE_2,PGF_(2a),TNF-α, IL-6,IL-1,TNF-α,IL-6,IL-1 and 5-HT.This research provides experimental data for clinical application of treating PD by electro acupuncture and a basis for further study.
There is abundant experience on PD treatment under TCM.To use the acupuncture and moxibustion to treat PD is very special and strong.It is cheaper,easier,non-poison etc.
引文
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