蒙医温针对疲劳大鼠作用复杂机制研究
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摘要
慢性疲劳综合征(Chronic Fatigue Syndrome, CFS)是一种以原因不明的、疲劳难以恢复为主要特征的症候群,可伴有长期低热、咽痛、头痛、肌肉关节肿痛、抑郁、健忘、失眠或嗜睡等症状。蒙医学认为CFS是由人体身、心、语活动异常状态的延续,在这种长期的异常状态中,机体一直处于一种慢性应激状态,使体内的免疫系统常处于慢性免疫激活状态,导致神经内分免疫系统调节功能紊乱,从而引出的CFS的一系列证状。
本课题从蒙医温针疗法调节疲劳大鼠复杂机制为切入点,从神经免疫水平研究CFS的致病机理和蒙医温针疗法的调节机制,阐明蒙医温针疗法抗疲劳的可能机理。
研究目的
探讨蒙医温针对疲劳大鼠的作用及调节复杂机制。
研究方法
SD雄性大鼠,体重160-180g,共60只,随机分为3组,即正常对照组、模型组和模型温针组,每组20只大鼠。
正常对照组：自造模开始3周,每日定点喂食2次,每次1小时,不予任何刺激。模型组：适应环境3天之后,开始造模,共21天。模型温针组：造模的同时,取蒙医“顶会”、“命脉穴”和“心穴”(隔2天取一个穴位),用特制银针针刺后,接MLY-I型蒙医疗术温针仪。电流强度100 mA,温度40℃,每次刺激15 min,每隔2d治疗1次,共7次。通过行为学检测对模型进行判定；检测大鼠血清TNF-α、ACTH、C的含量和下丘脑NE、5-HT、DA的含量及肝自由基的含量。
结果
1、造模前各组大鼠体重无显著性差异,造模后模型组与正常组比较有显著性差异,而温针组在于中间。表明应激所致,大鼠体重增长得缓慢。模型组比正常组行走个数和站立次数均减少。温针组与模型组相比,行走个数和站立次数均增多。表明,模型组大鼠出现身心疲劳状况。而温针可以改善模型大鼠的中枢神经系统的“疲劳”状态情绪倾向。从而有效调整躯体和心理疲劳状态。
2、模型组与对照组相比,TNF-α、ACTH和皮质酮含量均升高,有显著性差异。温针组与模型组相比,TNF-α、ACTH和皮质酮含量降低,有显著性差异。结果表明疲劳大鼠TNF-α、ACTH和皮质酮明显增高,提示内分泌免疫功能出现紊乱,而温针可以降低上述指标,使失调的内分泌免疫功能恢复正常。
3、与正常对照组相比,模型组NE、5-HT和DA含量升高,有显著性差异。与模型组相比,温针组NE、5-HT和DA含量降低,有显著性差异。
4、与正常对照组相比,模型组MDA含量有增高的趋势,但是无显著性差异。与模型组相比,温针组MDA含量有下降的趋势,但是无显著性差异。与正常对照组相比,模型组GSH含量降低,有显著性差异。与模型组相比,温针组GSH含量增高,有显著性差异。与正常对照组相比,模型组GSH-PX含量降低,有显著性差异。与模型组相比,温针组GSH-PX含量增高,有显著性差异。与正常对照组相比,模型组SOD含量降低,有显著性差异。与模型组相比,温针组SOD含量增高,有显著性差异。
结论
1、以往的蒙医学研究把CFS病因归纳为“巴达干-赫依”或“赫依”,而本研究首次提出本病的病因为“由于身、心、语功能过度诱发体内三根失调而致CFS”,的观点。
2、疲劳模型的制作方法很多。如跑步、捆绑和游泳等。本项研究基于长期三根失调而发CFS的蒙医理论,采用力竭游泳和束缚方法制作了疲劳模型。研究表明游泳比跑步能消耗体力。强制性游泳能引起动物绝望等心理压力,从而更能消耗体力。从而强制性游泳加束缚刺激而制作的疲劳模型更符合蒙医身心语功能过度而致CFS的观点。
3、复合应激所致的疲劳模型大鼠血清、下丘脑和肝组织的各种观察指标均出现不同程度的改变。而这些经典指标为引起疲劳的主要原因。这与精华与糟粕分离失衡而诱发体内三根失调导致CFS的蒙医观点不谋而合。实验结果表明,蒙医温针通过调节疲劳大鼠血清、下丘脑和肝组织的各种观察指标的变化,而起抗疲劳作用。
4、本实验所选用的蒙医顶会穴、命脉穴和心穴等三穴主要用于调节三根失调、调体素等。从临床作用来看也符合CFS的治疗需求。表明CFS以三根失调而发病,其中以赫依为甚,并与心脏功能关系密切。
5、蒙医温针疗法主要以调节赫依巴达干的功能失调、改善气血循行、提高机体免疫功能,激发调动和增强机体的抗病能力等作用对CFS起到良好的抗疲劳作用。并具有操作简便、安全、有效、毒副作用小等特点和优势。值得广泛推广应用。
Chronic Fatigue Syndrome(CFS) is a unexplained syndrome mainly characterized by rave recoverable fatigue that are associated with multiple symptoms such as long term low fever,sore throat、headache、muscle and joint swelling and pain、blahs、forgetting、insomnia and sleepiness. In the Mongolian Medicine CFS was thought to the caused by physical, feeling (thought) and language activities of human body in the abnormal condition for long term.In this (disease) progression there are accumulating stress in the body, result in disorders of the nervous, immune and endocrine functions and lead to present a series of symptoms of CFS. This project focus on mechanisms the effects of Mongolian Medical warm Acupuncture therapy (MMWAT) on CFS,investigate the anti-fatigue effect and its regulate mechanisms on the fatigue in rat model using exhaustive swimming test plus tied methods and behavioral tests.
OBJECTIVE
To exposure the effects and its mechanisms of the Mongolian Medical Warm Acupuncture therapy on fatigue in rat modal.
METHODS
SD male rat, weigh 160-180g,60 rats are into 3 groups:normal control group (NC)、fatigue modal group (FM) and warm acupuncture group (WAT); 20 rats in every group rats are feed by daily fixed feed stuff two times, feeding forl hour. before beginning of experiments.
NC group:no any stimulation.
FM group:Fatigue rat model was made by using exhaustive swimming plus tied methods to stimulate rats for 3 weeks.
WAT group:do the same modeling process with Mongolian warm Acupuncture therapy using adding 100m A currents and 40℃temperature to the handtailor silver acupuncture needles through MLY-1 type Mongolian medical treatment method warm-pin Instrument an the selected Mongolian medical points-"Dinghui ", " Mingmai "and "Xinxue" (treat one time on one point in very 2d (day), earth stimulus (treatment) lasted for 15min, a total of seven times in 3 weeks). Fatigue mode; and effects of warm acupuncture therapy was evaluated by the ethalogy test, examine TNF-a, ACTH, C content in rat serum, NE、5-HT、DA in rat hyothalamus and free radical group in rat liver.
RESULTS
1.On rat body weight,there are no significant difference among the 3 groups before modeling experiments:end of the modeling experiments, there are significant difference in the between FM group and NC group, FM group rats lost body weight; body weight of WAT group rats are in the between of MF group and NC group.showed that stress induce loss of body weight in MF group rat. MF group rat compared with NC group, the number of walking and standing activities were increased. Shows that MF group rat presents physical and mental fatigue, warm acupuncture therapy can relief the stress and improve the physical and mental fatigue.
In the modeling of 7d, struggling times of rats have no significant difference among the 3groups.In the modeling of 14d, MF group and WAT group rats compared with NC group respectively, the struggling times were increased and have significant differences. In the modeling of 21d, the model group、the warm acupuncture group compared with the comparison group, struggling times decreased,there are significant differences.In the modeling of 7d,14d,21d, the warm acupuncture group compared with the model group, exhaustive swimming time was significantly prolonged,there are significant differences.
2.The MF group rats compared with NC group, content of TNF-α、ACTH and Corticosterone were in elevated and have significant differences.The MAT group RATS compared with the comparison with NC group, content of TNF-α、ACTH and Corticosterone content were reduced and have significant differences. The results indicate that the TNF-a, ACTH and Corticosterone in fatigue rats were significantly increased, suggesting that endocrine and immune function present disorders, and the warm Acupuncture therapy can improve the endocrine and immune function disorders.
3. Compared with NC group, contents of NE、5-HT and DA in MF group rat were increased and have significant differences. Compared with the MF group, contents of NE、5-HT and DA in WAT group rats were reduced and have significant differences. These the result suggests that nervous function disorders are involved in the pathogenesis of CFS (fatigue) and warm acupuncture therapy can improve the nervous functions disorders.
4. Compared with NC group, MDA content in MF group rats, tend to increase, but there are no significant differences.Compared with MF group, MDA content in WAT group rats tend to decrease, but there are also no significant differences. Compared with NC group,GSH content in MF group rats were increased and have significant differences Compared with NC group, GSH-PX content in MF group rats were increased significant differences. Compared with MF group, GSH-PX content in MAT group rats were increased and have significant differences;
CONCLUSIONS AND DISCUSSION
1.Previous Mongolian medical research described that CFS was caused by abnormal "Badagan" or "Hei", The present study first time exposure a new mechanisms of CFS onset that CFS was caused by the disorders of "three GenSu " that were triggered by abnormal "physical, thought and language" activities in body for long term.
2. There are several ways to make the model of fatigue. Such as running, tied, and swimming. According to Mongolian medical theory, we proposed that CFS should be caused by disorders of " the three GenSu" that CFS should be triggered by abnormal physical, thought and language activities in body for long term,and that the Mongolian warm Acupuncture therapy should have a anti-fatigue effects. Therefore, we designed a alternative pathological model for CFS onset in rat that was combination exhaustive swimming and tied methods to induce fatigue.Studies have shown that swimming was more exhaustive than running. Mandatory swimming can induce animals despair and other psychological pressure, and more exhaustion. Therefore a mandatory swimming plus tied to stimulate and make fatigue model is more suitable the Mongolian medical theory and our proposal.
3. In this study, long term stimulation of rats with exhaustive swimming plus tied methods induce abnormal physical, feeling activities, result in accumulation of stress. Accumulation of stress could induce disorders of nervous, endocrine and immune functions and lead to cause fatigue. These are well consistent with our proposal that CFS should be caused by the disorders of "the three GenSu" that were triggered by abnormal physical,though (feeling)and language activities. our proposal were also well supported by the results that Mongolian warm acupuncture therapy can improve the disorders of nervous, endocrine and immune function and relief the fatigue.
4.DinghuiXue、MaiyongXue and XinXue 3 points that were selected for this experiment used to regulate the disorders of " the three GenSu". Clinical usage of the 3 points are well suitable for treatment of CFS (fatigue), indicate that CFS onset was due to the disorders of" the three GenSu". These further supported our proposal.
5. Mongolian warm acupuncture therapy can regulate the disorders of " the three GenSu" and improve the disorders of nervous,endocrine and immune functions, and also improve the ability to defence of disease so that have a well anti-fatigue an CFS. This might be a possible mechanism underlying the anti-fatigue effects of Mongolian warm acupuncture therapy on CFS treatment.
6. Well anti-fatigue effects of Mongolian warm acupuncture therapy could provide a new potential therapeutic and technique for clinical treatment of CFS .Moreover, Mongolian warm acupuncture therapy had many advantages and features such as simple, safety, effective and less toxic and side effects etc. Therefore, this study strongly recommend Mongolian warm acupuncture therapy to widely use in clinical treatment.
本课题从蒙医温针疗法调节疲劳大鼠复杂机制为切入点,从神经免疫水平研究CFS的致病机理和蒙医温针疗法的调节机制,阐明蒙医温针疗法抗疲劳的可能机理。
研究目的
探讨蒙医温针对疲劳大鼠的作用及调节复杂机制。
研究方法
SD雄性大鼠,体重160-180g,共60只,随机分为3组,即正常对照组、模型组和模型温针组,每组20只大鼠。
正常对照组：自造模开始3周,每日定点喂食2次,每次1小时,不予任何刺激。模型组：适应环境3天之后,开始造模,共21天。模型温针组：造模的同时,取蒙医“顶会”、“命脉穴”和“心穴”(隔2天取一个穴位),用特制银针针刺后,接MLY-I型蒙医疗术温针仪。电流强度100 mA,温度40℃,每次刺激15 min,每隔2d治疗1次,共7次。通过行为学检测对模型进行判定；检测大鼠血清TNF-α、ACTH、C的含量和下丘脑NE、5-HT、DA的含量及肝自由基的含量。
结果
1、造模前各组大鼠体重无显著性差异,造模后模型组与正常组比较有显著性差异,而温针组在于中间。表明应激所致,大鼠体重增长得缓慢。模型组比正常组行走个数和站立次数均减少。温针组与模型组相比,行走个数和站立次数均增多。表明,模型组大鼠出现身心疲劳状况。而温针可以改善模型大鼠的中枢神经系统的“疲劳”状态情绪倾向。从而有效调整躯体和心理疲劳状态。
2、模型组与对照组相比,TNF-α、ACTH和皮质酮含量均升高,有显著性差异。温针组与模型组相比,TNF-α、ACTH和皮质酮含量降低,有显著性差异。结果表明疲劳大鼠TNF-α、ACTH和皮质酮明显增高,提示内分泌免疫功能出现紊乱,而温针可以降低上述指标,使失调的内分泌免疫功能恢复正常。
3、与正常对照组相比,模型组NE、5-HT和DA含量升高,有显著性差异。与模型组相比,温针组NE、5-HT和DA含量降低,有显著性差异。
4、与正常对照组相比,模型组MDA含量有增高的趋势,但是无显著性差异。与模型组相比,温针组MDA含量有下降的趋势,但是无显著性差异。与正常对照组相比,模型组GSH含量降低,有显著性差异。与模型组相比,温针组GSH含量增高,有显著性差异。与正常对照组相比,模型组GSH-PX含量降低,有显著性差异。与模型组相比,温针组GSH-PX含量增高,有显著性差异。与正常对照组相比,模型组SOD含量降低,有显著性差异。与模型组相比,温针组SOD含量增高,有显著性差异。
结论
1、以往的蒙医学研究把CFS病因归纳为“巴达干-赫依”或“赫依”,而本研究首次提出本病的病因为“由于身、心、语功能过度诱发体内三根失调而致CFS”,的观点。
2、疲劳模型的制作方法很多。如跑步、捆绑和游泳等。本项研究基于长期三根失调而发CFS的蒙医理论,采用力竭游泳和束缚方法制作了疲劳模型。研究表明游泳比跑步能消耗体力。强制性游泳能引起动物绝望等心理压力,从而更能消耗体力。从而强制性游泳加束缚刺激而制作的疲劳模型更符合蒙医身心语功能过度而致CFS的观点。
3、复合应激所致的疲劳模型大鼠血清、下丘脑和肝组织的各种观察指标均出现不同程度的改变。而这些经典指标为引起疲劳的主要原因。这与精华与糟粕分离失衡而诱发体内三根失调导致CFS的蒙医观点不谋而合。实验结果表明,蒙医温针通过调节疲劳大鼠血清、下丘脑和肝组织的各种观察指标的变化,而起抗疲劳作用。
4、本实验所选用的蒙医顶会穴、命脉穴和心穴等三穴主要用于调节三根失调、调体素等。从临床作用来看也符合CFS的治疗需求。表明CFS以三根失调而发病,其中以赫依为甚,并与心脏功能关系密切。
5、蒙医温针疗法主要以调节赫依巴达干的功能失调、改善气血循行、提高机体免疫功能,激发调动和增强机体的抗病能力等作用对CFS起到良好的抗疲劳作用。并具有操作简便、安全、有效、毒副作用小等特点和优势。值得广泛推广应用。
Chronic Fatigue Syndrome(CFS) is a unexplained syndrome mainly characterized by rave recoverable fatigue that are associated with multiple symptoms such as long term low fever,sore throat、headache、muscle and joint swelling and pain、blahs、forgetting、insomnia and sleepiness. In the Mongolian Medicine CFS was thought to the caused by physical, feeling (thought) and language activities of human body in the abnormal condition for long term.In this (disease) progression there are accumulating stress in the body, result in disorders of the nervous, immune and endocrine functions and lead to present a series of symptoms of CFS. This project focus on mechanisms the effects of Mongolian Medical warm Acupuncture therapy (MMWAT) on CFS,investigate the anti-fatigue effect and its regulate mechanisms on the fatigue in rat model using exhaustive swimming test plus tied methods and behavioral tests.
OBJECTIVE
To exposure the effects and its mechanisms of the Mongolian Medical Warm Acupuncture therapy on fatigue in rat modal.
METHODS
SD male rat, weigh 160-180g,60 rats are into 3 groups:normal control group (NC)、fatigue modal group (FM) and warm acupuncture group (WAT); 20 rats in every group rats are feed by daily fixed feed stuff two times, feeding forl hour. before beginning of experiments.
NC group:no any stimulation.
FM group:Fatigue rat model was made by using exhaustive swimming plus tied methods to stimulate rats for 3 weeks.
WAT group:do the same modeling process with Mongolian warm Acupuncture therapy using adding 100m A currents and 40℃temperature to the handtailor silver acupuncture needles through MLY-1 type Mongolian medical treatment method warm-pin Instrument an the selected Mongolian medical points-"Dinghui ", " Mingmai "and "Xinxue" (treat one time on one point in very 2d (day), earth stimulus (treatment) lasted for 15min, a total of seven times in 3 weeks). Fatigue mode; and effects of warm acupuncture therapy was evaluated by the ethalogy test, examine TNF-a, ACTH, C content in rat serum, NE、5-HT、DA in rat hyothalamus and free radical group in rat liver.
RESULTS
1.On rat body weight,there are no significant difference among the 3 groups before modeling experiments:end of the modeling experiments, there are significant difference in the between FM group and NC group, FM group rats lost body weight; body weight of WAT group rats are in the between of MF group and NC group.showed that stress induce loss of body weight in MF group rat. MF group rat compared with NC group, the number of walking and standing activities were increased. Shows that MF group rat presents physical and mental fatigue, warm acupuncture therapy can relief the stress and improve the physical and mental fatigue.
In the modeling of 7d, struggling times of rats have no significant difference among the 3groups.In the modeling of 14d, MF group and WAT group rats compared with NC group respectively, the struggling times were increased and have significant differences. In the modeling of 21d, the model group、the warm acupuncture group compared with the comparison group, struggling times decreased,there are significant differences.In the modeling of 7d,14d,21d, the warm acupuncture group compared with the model group, exhaustive swimming time was significantly prolonged,there are significant differences.
2.The MF group rats compared with NC group, content of TNF-α、ACTH and Corticosterone were in elevated and have significant differences.The MAT group RATS compared with the comparison with NC group, content of TNF-α、ACTH and Corticosterone content were reduced and have significant differences. The results indicate that the TNF-a, ACTH and Corticosterone in fatigue rats were significantly increased, suggesting that endocrine and immune function present disorders, and the warm Acupuncture therapy can improve the endocrine and immune function disorders.
3. Compared with NC group, contents of NE、5-HT and DA in MF group rat were increased and have significant differences. Compared with the MF group, contents of NE、5-HT and DA in WAT group rats were reduced and have significant differences. These the result suggests that nervous function disorders are involved in the pathogenesis of CFS (fatigue) and warm acupuncture therapy can improve the nervous functions disorders.
4. Compared with NC group, MDA content in MF group rats, tend to increase, but there are no significant differences.Compared with MF group, MDA content in WAT group rats tend to decrease, but there are also no significant differences. Compared with NC group,GSH content in MF group rats were increased and have significant differences Compared with NC group, GSH-PX content in MF group rats were increased significant differences. Compared with MF group, GSH-PX content in MAT group rats were increased and have significant differences;
CONCLUSIONS AND DISCUSSION
1.Previous Mongolian medical research described that CFS was caused by abnormal "Badagan" or "Hei", The present study first time exposure a new mechanisms of CFS onset that CFS was caused by the disorders of "three GenSu " that were triggered by abnormal "physical, thought and language" activities in body for long term.
2. There are several ways to make the model of fatigue. Such as running, tied, and swimming. According to Mongolian medical theory, we proposed that CFS should be caused by disorders of " the three GenSu" that CFS should be triggered by abnormal physical, thought and language activities in body for long term,and that the Mongolian warm Acupuncture therapy should have a anti-fatigue effects. Therefore, we designed a alternative pathological model for CFS onset in rat that was combination exhaustive swimming and tied methods to induce fatigue.Studies have shown that swimming was more exhaustive than running. Mandatory swimming can induce animals despair and other psychological pressure, and more exhaustion. Therefore a mandatory swimming plus tied to stimulate and make fatigue model is more suitable the Mongolian medical theory and our proposal.
3. In this study, long term stimulation of rats with exhaustive swimming plus tied methods induce abnormal physical, feeling activities, result in accumulation of stress. Accumulation of stress could induce disorders of nervous, endocrine and immune functions and lead to cause fatigue. These are well consistent with our proposal that CFS should be caused by the disorders of "the three GenSu" that were triggered by abnormal physical,though (feeling)and language activities. our proposal were also well supported by the results that Mongolian warm acupuncture therapy can improve the disorders of nervous, endocrine and immune function and relief the fatigue.
4.DinghuiXue、MaiyongXue and XinXue 3 points that were selected for this experiment used to regulate the disorders of " the three GenSu". Clinical usage of the 3 points are well suitable for treatment of CFS (fatigue), indicate that CFS onset was due to the disorders of" the three GenSu". These further supported our proposal.
5. Mongolian warm acupuncture therapy can regulate the disorders of " the three GenSu" and improve the disorders of nervous,endocrine and immune functions, and also improve the ability to defence of disease so that have a well anti-fatigue an CFS. This might be a possible mechanism underlying the anti-fatigue effects of Mongolian warm acupuncture therapy on CFS treatment.
6. Well anti-fatigue effects of Mongolian warm acupuncture therapy could provide a new potential therapeutic and technique for clinical treatment of CFS .Moreover, Mongolian warm acupuncture therapy had many advantages and features such as simple, safety, effective and less toxic and side effects etc. Therefore, this study strongly recommend Mongolian warm acupuncture therapy to widely use in clinical treatment.
引文
[1] Holmes GP, Kaplan JE, Gnatz NM, et al. Chronic Fatigue Syndrome: A Working Case Definition. Ann Intern Med 1988,108:387-389
[2] Schluederberg A. NIH Coference: Chronic Fatigue Syndrome, Research Ann Intern Med} 1992,117(4): 325-331
[3] Fukuda. Complete Text of Revised Case Definition. Annals of Internal Medicine, 1994,121(12): 953-959
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