逆灸对力竭大鼠Ghrelin的影响及能量代谢相关机制的研究
摘要
“逆针灸”是中医“治未病”的重要手段之一,具有防病治病、保健延衰的作用。“逆灸”是其中最为常用和易用的方式。我们通过对逆灸现代研究的学习,发现逆灸对机体的调节作用非常广泛,涉及到大量基础的指标,具有良性、广泛性和潜伏性的作用特点。中医理论认为逆灸通过扶助正气、增强身体素质,从而增强机体对抗疾病的能力。我们认为正气的核心是机体的自稳调节能力。艾灸适宜的刺激是一种良性应激原,提前给予艾灸处理,能够使机体产生适度预应激,能够针对潜隐性的功能紊乱进行调整,提高机体的自稳调节能力,使机体的应激耐受力增强,以抵抗随后过度应激对机体的伤害。
我们前期实验观察到逆灸可以减轻随后的“疾病状态”大鼠(佐剂性关节炎、痛经模型)的发病率与疾病程度,也对“亚健康状态”雌性大鼠机体(更年期模型)有良性的调节作用。我们想进一步观察逆灸对“健康”机体的作用。结合逆灸的作用特点,我们需要一个能体现机体综合素质的指标,以全面概括逆灸的“良性预应激”作用。
运动能力是动物的基本能力之一,体现机体的生命力,需要各个系统的全面配合,是机体综合素质的体现。强大的运动能力可以说是机体整体健康程度的一种外在表现,也可以说是机体“正气充足”的体现。我们可以通过观察逆灸对动物运动耐力的影响,进而推测逆灸是如何通过“扶助”正常机体的“正气”而达到防病保健的效果。极限运动耐力属于运动能力的一种,可以使机体将自身潜力充分发挥出来,更能体现出机体的生命潜力。同时,极限耐力运动属于过度应激,与逆灸的适度应激相对应,两者一前一后可以体现出两种应激叠摞后的效应。
能量代谢是动物所有功能活动的基础,应激是机体内环境面对自身及外环境的变动而产生的变化,两者的稳定对维持整个机体的稳定具有重要意义。极限耐力运动需要持续的能量供应,同时也是一种会造成机体过度应激反应的不良应激原,因此极限耐力运动能够很好地将应激与能量代谢联系起来。Ghrelin由于其特殊的脑肠肽特性,可能在应激与能量代谢系统甚至其他系统之间的稳态调节都起到关键因子的作用。应激、能量代谢与Ghrelin水平三者可能互相影响,互相调节,共同维持着机体的稳态。我们认为这些指标能够反映机体的自稳调节能力,可以通过观察这些指标在极限耐力运动中的变化探讨逆灸对身体素质的改善作用。
关元穴和命门穴是传统保健穴,联系人体的元气,作用比较基础、广泛,并且在穴性方面比较有代表性(一阴一阳)。选用两穴进行比较,一方面可以体现逆灸的共同效应,一方面又能反映逆灸不同穴位的特异性。
目的：通过观察逆灸关元穴和逆灸命门穴对大鼠极限运动耐力的提升作用,探讨逆灸的扶助正气、增强身体素质的效果。通过观察逆灸、力竭和逆灸+力竭对大鼠肝糖原、血乳酸、心肌和血清NOS、 HPA轴应激激素、HPG轴相关激素、下丘脑和血清Ghrelin水平等指标的影响,探讨逆灸对力竭运动大鼠Ghrelin的调节作用及其与能量代谢的相关性,进而探究逆灸提高机体运动耐力的部分机制,进一步揭示逆灸防病保健的科学内涵,或为运动医学提供参考。
方法：将48只SD雄性大鼠分为：空白对照组、逆灸关元组、逆灸命门组、力竭对照组、逆灸关元+力竭组和逆灸命门+力竭组。逆灸关元组、逆灸命门组、逆灸关元+力竭组和逆灸命门+力竭组分别行关元穴和命门穴艾条灸法,隔天1次,每次10min,连续10次；空白对照组和力竭对照组与上述组同时人工抓取但不灸,如此连续10次。第20天,即最后一次艾灸24h后,力竭对照组、逆灸关元+力竭组和逆灸命门+力竭组混合后随机抽取进行温水力竭游泳并记录游泳至力竭的时间。力竭即刻与空白对照组、逆灸关元组和逆灸命门组一同处死取材。观察逆灸关元穴和命门穴对大鼠力竭游泳时间的影响。采用生化法检测各组大鼠肝糖原、血乳酸、心肌和血清NOS活性、HPA轴应激激素水平(下丘脑CRH、垂体ACTH血清CORT)、HPG轴相关激素(下丘脑GnRH、血清睾酮)、中枢及外周Ghrelin水平。
结果：
1力竭时间
逆灸关元+力竭组和逆灸命门+力竭组的游泳力竭平均时间都明显长于力竭对照组(P<0.01,P<0.05)；两个逆灸+力竭组之间无显著差异(P>0.05)。
2肝糖原含量
逆灸关元组和逆灸命门组的肝糖原含量与空白对照组相比没有显著变化(P>0.05)。力竭对照组的肝糖原含量较空白对照组极显著减少(P<0.01)。逆灸关元+力竭组与逆灸命门+力竭组的肝糖原较力竭对照组均有极显著或显著升高(P<0.01,P<0.05)。两个逆灸+力竭组之间无显著差异(P>0.05)。
3血乳酸含量
逆灸关元组和逆灸命门组的血乳酸含量与空白对照组相比均没有明显变化(P>0.05)。力竭对照组的血乳酸较空白对照组极显著升高(P<0.01)。逆灸关元+力竭组与逆灸命门+力竭组的血乳酸与力竭对照组相比均有极显著的降低(P<0.01)。两个逆灸+力竭组之间无差异(P>0.05)。
4心肌和血清NOS活性
与空白对照组相比,逆灸关元组和逆灸命门组的心肌NOS活性没有明显变化(P>0.05),力竭对照组的心肌NOS活性极显著升高(P<0.01)；与力竭对照组比较,逆灸关元+力竭组心肌NOS活性没有明显变化(P>0.05),逆灸命门+力竭组心肌NOS活性极显著下降(P<0.01)。两个逆灸+力竭组之间有显著统计学差异(P<0.05)。
各组血清NOS活性和心肌NOS活性的变化情况一致。
5下丘脑CRH含量
与空白对照组比较,逆灸关元穴组和逆灸命门穴组大鼠的下丘脑CRH含量均无明显变化(P>0.05)。力竭对照组大鼠的下丘脑CRH含量较空白对照组有极显著升高(P<0.01)。与力竭对照组比较,逆灸关元+力竭组大鼠的下丘脑CRH含量没有变化(P>0.05)；逆灸命门+力竭组大鼠的下丘脑CRH含量极显著降低(P<0.01)。两个逆灸+力竭组之间有极显著统计学差异(P<0.01)。
6垂体ACTH含量
与空白对照组比较,逆灸关元穴组和逆灸命门穴组大鼠的垂体ACTH含量均无明显变化(P>0.05)。力竭对照组大鼠的垂体ACTH含量较空白对照组有极显著升高(P<0.01)。与力竭对照组比较,逆灸关元+力竭组大鼠的垂体ACTH含量没有变化(P>0.05)；逆灸命门+力竭组大鼠的垂体ACTH含量极显著降低(P<0.01)。两个逆灸+力竭组之间有极显著统计学差异(P<0.01)。
7血清CORT含量
与空白对照组比较,逆灸关元穴组和逆灸命门穴组大鼠的血清CORT含量均无明显变化(P>0.05)。力竭对照组大鼠的血清CORT含量较空白对照组有极显著升高(P<0.01)。与力竭对照组比较,逆灸关元+力竭组大鼠的血清CORT含量没有变化(P>0.05)；逆灸命门+力竭组大鼠的血清CORT含量极显著降低(P<0.01)。两个逆灸+力竭组之间有极显著统计学差异(P<0.01)。
8下丘脑GnRH含量
与空白对照组相比,逆灸命门穴组大鼠的下丘脑GnRH含量无明显变化(P>0.05),力竭对照组大鼠的下丘脑GnRH极显著升高(P<0.01)。与力竭对照组比较,逆灸关元+力竭组大鼠下丘脑GnRH含量没有明显变化(P>0.05)；逆灸命门+力竭组大鼠下丘脑GnRH含量极显著降低(P<0.01)。两个逆灸+力竭组之间的下丘脑GnRH含量有显著统计学差异(P<0.05)。
9血清睾酮含量
与空白对照组相比,逆灸命门穴组大鼠的血清睾酮含量无明显变化(P>0.05),力竭对照组大鼠的血清睾酮极显著升高(P<0.01)。与力竭对照组比较,逆灸关元+力竭组大鼠血清睾酮含量没有明显变化(P>0.05)；逆灸命门+力竭组大鼠血清睾酮含量极显著降低(P<0.01)。两个逆灸+力竭组之间的血清睾酮含量差异有显著统计学意义(P<0.05)。
10下丘脑和血清Ghrelin含量
与空白对照组相比,逆灸命门穴组大鼠的下丘脑Ghrelin含量无明显变化(P>0.05),力竭对照组大鼠的下丘脑Ghrelin含量极显著升高(P<0.01)。与力竭对照组比较,逆灸关元+力竭组大鼠下丘脑Ghrelin含量没有明显变化(P>0.05)；逆灸命门+力竭组大鼠下丘脑Ghrelin含量极显著降低(P<0.01)。两个逆灸+力竭组之间的下丘脑Ghrelin含量有显著统计学差异(P<0.05)。
各组血清Ghrelin和下丘脑Ghrelin含量的变化情况一致。
结论：
1逆灸对机体是一种适度的应激,对机体的调节作用不会预先在单个生化指标上体现出来。但却能显著改善机体的健康程度,提高机体对不良应激的抵抗力。
2力竭运动是一种能够引起机体过度应激反应的不良应激原。力竭导致机体应激轴过度激活,过分调动了能源储备,能量代谢处于紊乱状态。
3我们认为“正气”的核心是机体的“自稳调节能力”。逆灸正是通过提高机体的自稳调节能力以扶助正气,进而增强机体素质,提高运动耐力。这可能能够为运动医学提供一定的参考。其具体机制可能与调节应激激素,能量代谢相关因子和Ghrelin水平等有关。
4关元穴与命门穴的效应有所不同。关元穴在延长大鼠游泳时间、调节糖原储备和乳酸代谢方面与命门穴无异,但对应激激素、性激素、NOS活性及Ghrelin水平的调节效应不明显,关元穴对这些指标的效应可能具有滞后性,也可能其对大鼠体能的提升作用以及对能量代谢的调节作用是通过其他途径实现的。两穴差别有待我们进一步研究探讨。
Preventive acupuncture and moxibustion are important methods of treating pre-disease in traditional Chinese medicine, which have played an important role in disease prevention, treatment, health care and life extension. According to recent researches, the regulation effect of preventive mexibustion is extensive and involving many fundamental physiological processes. The features of preventive moxibustion can be summarized into three traits:beneficial, extensive, and latent. According to the theory of traditional Chinese medicine, preventive moxibustion enhance the anti-disease ability by building up the health energy (Zheng Qi). We hold that self-rebalance ability is the core part of health energy (Zheng Qi). As a benign stressor giving mild pre-stress to the body, moxibustion will enhance the self-rebalance ability and correct the potential dysfunctions to enable the body to mobilize more potential abilities to resist the damages and illnesses causing by the following over-stress. In our latest experiment, we observed that preventive moxibustion could reduce incidence and severity of disease (adjuvant arthritis, dysmenorrhea model) and give benign regulation to "sub-health" state (menopausal model) in modeled rats. Therefore, we are going to observe the effect of preventive moxibustion on healthy state of animals. Considering the complexity of the comprehensive effect, we need a physiological index that can be representative for the comprehensive quality of the body and the "benign pre-stress" effect of preventive moxibustion. Locomotivity is one of the basic abilities representing the vitality of the body. According to physiology studies, locomotivity depend on the ensemble coordinating operation of all the organs and systems of the body. According to the theory of traditional Chinese medicine, strong locomotivity means strong health energy (Zheng Qi), which is the comprehensive representation of the health condition of the human body. Through observing the effect of preventive moxibustion on locomotivity of animals, we can know how the body will be healthier by strengthening the health energy (Zheng Qi) with preventive moxibustion. As a kind of locomotivity, extreme exercise endurance, being able to force the body to fully perform its potential, is representative of the comprehensive quality of the body. Meanwhile, being a harmful over-stressor that will lead to excessive stress, extreme exercise endurance can be paired with preventive moxibustion in the comparison of the effect of benign pre-stress and harmful over-stress.
Energy metabolism is fundamental for all the functional activity of animal; and stress is the change of internal environment to external environment. The stability of energy metabolism and stress is important in the maintaining of the stability of the whole body. Being a harmful stressor, extreme exercise will not only cause over-stress response but also needs continuous supply of energy, therefore it can be a link between stress and energy metabolism. As a brain-gut peptide, ghrelin may be playing a key role in regulating the balance between energy metabolism and stress response or even other systems. Stress, energy metabolism and ghrelin level may have influence on each other in the keeping of the homeostasis. We hold that these index can represent the self-rebalance ability of the body, and their changes in exhaustive exercise can be useful in the study of effect of preventive moxibustion on the health of the body. Guanyuan(RN-4) and Mingmen(DU-4)point point are the most commonly used points in health keeping because of their extensive effects and their opposite nature in Yin-Yang. Comparing the two point in the experiment not only can represent the common effect of moxibustion but also show the deviation effect of different acu-points.
Objective:To discuss the enhancement of health energy and improvement of constitution with preventive moxibustion through observing the effect of preventive moxibustion at Guanyuan and Mingmen points on exercise endurance of rats. To discuss the correlation and influence of preventive moxibustion on ghrelin level and energy metabolism of exhaustive swimming rats through observation of the changes of liver glycogen, serum lactate, myocardium NOS, serum NOS, hormone of HPA axis, hormone of HPG axis, hypothalamus ghrelin and serum ghrelin before and after preventive moxibustion and exhausted swimming. And also introduce preventive moxibustion to sports medicine.
Methods:Forty-eight male SD rats were divided into six groups:normal control group, Guanyuan moxibustion group, Mingmen moxibustion group, exhaustion control group, Guanyuan moxibustion+exhaustion group and Mingmen moxibustion+exhaustion group. All moxibustion groups received a ten-minute moxibustion every two days, ten times in all. The two control groups received sham treatment at the same time with moxibustion groups. In the twentith day, twenty four hours after the last moxibustion, exhaustion control group, Mingmen moxibustion+exhaustion group and Guanyuan moxibustion+exhaustion group received a warm water exhaustive swimming and the swimming exhausted time was recorded. After swimming exhaustion, all rats were executed and sampled whole blood, right lobe of liver, heart muscle, hypothalamus, hypophysis and right side adrenal gland. The swimming time of all swimming groups were compared. Glycogen level in liver, lactate level in serum, NOS level in myocardium, NOS level in serum, hormone level of HPA axis, hormone level of HPG axis, ghrelin level in hypothalamus and ghrelin level in serum were detected.
Results:
1Swimming exhausted time
Compared to the exhaustion control group, the swimming exhausted time of Guanyuan moxibustion+exhaustion group and Mingmen moxibustion+exhaustion group were significantly higher(P<0.01, P＜0.05), there was no difference between the two moxibustion+exhaustion groups (P>0.05)
2Glycogen level in liver
Compared to the normal control group, the glycogen level in liver of Guanyuan moxibustion group and Mingmen moxibustion group had no significant changes (P>0.05), that of the exhaustion control group significantly decreased (P＜0.01).
Compared to the exhaustion control group, the glycogen level in liver of Guanyuan moxibustion+exhaustion group and Mingmen moxibustion+exhaustion group were significantly increased (P＜0.01, P<0.05), and there was no difference between the two moxibustion+exhaustion groups (P>0.05).
3lactate level in blood
Compared to the normal control group, the lactate level in blood of Guanyuan moxibustion group and Mingmen moxibustion group had no significant changes (P>0.05), that of the exhaustion control group significantly increased (P<0.01).
Compared to the exhaustion control group, the lactate level in blood of Guanyuan moxibustion+exhaustion group and Mingmen moxibustion+exhaustion group were significantly dropped (P＜0.01), and there was no difference between the two moxibustion+exhaustion groups (P>0.05).
4NOS level in myocardium and serum
Compared to the normal control group, the NOS level in myocardium of Guanyuan moxibustion group and Mingmen moxibustion group had no significant changes (P>0.05), that of the exhaustion control group significantly increased (P＜0.01).
Compared to the exhaustion control group, the NOS level in myocardium of Mingmen moxibustion+exhaustion group was significantly decreased (P<0.01), but the change of Guanyuan moxibustion+exhaustion group was not significant (P>0.05), and there were significant differences between the two moxibustion+exhaustion groups (P＜0.05). The change pattern of NOS level in serum was the same as that in myocardium.
5CRH level in hypothalamus
Compared to the normal control group, the CRH level in hypothalamus of Guanyuan moxibustion group and Mingmen moxibustion group had no significant changes (P>0.05), that of the exhaustion control group significantly increased (P<0.01).
Compared to the exhaustion control group, the CRH level in hypothalamus of Mingmen moxibustion+exhaustion group was significantly decreased (P<0.01), but the change of Guanyuan moxibustion+exhaustion group was not significant (P>0.05), and there were significant differences between the two moxibustion+exhaustion groups (P<0.05).
6ACTH level in hypophysis
Compared to the normal control group, the ACTH level in hypophysis of Guanyuan moxibustion group and Mingmen moxibustion group had no significant changes (P>0.05), that of the exhaustion control group significantly increased (P<0.01).
Compared to the exhaustion control group, the ACTH level in hypophysis of Mingmen moxibustion+exhaustion group was significantly decreased (P<0.01), but the change of Guanyuan moxibustion+exhaustion group was not significant (P>0.05), and there were significant differences between the two moxibustion+exhaustion groups (P＜0.05).7Cortisol level in serum
Compared to the normal control group, the cortisol level in serum of Guanyuan moxibustion group and Mingmen moxibustion group had no significant changes (P>0.05), that of the exhaustion control group significantly increased (P<0.01).
Compared to the exhaustion control group, the cortisol level in serum of Mingmen moxibustion+exhaustion group was significantly decreased (P＜0.01), but the change of Guanyuan moxibustion+exhaustion group was not significant (P>0.05), and there were significant differences between the two moxibustion+exhaustion groups (P<0.01).
8GnRH level in hypothalamus
Compared to the normal control group, the GnRH level in hypothalamus of Guanyuan moxibustion group and Mingmen moxibustion group had no significant changes (P>0.05), that of the exhaustion control group significantly increased (P<0.01).
Compared to the exhaustion control group, the GnRH level in hypothalamus of Mingmen moxibustion+exhaustion group was significantly decreased (P<0.01), but the change of Guanyuan moxibustion+exhaustion group was not significant (P>0.05), and there were significant differences between the two moxibustion+exhaustion groups (P＜0.05).
9Testosterone level in serum
Compared to the normal control group, the testosterone level in serum of Guanyuan moxibustion group and Mingmen moxibustion group had no significant changes (P>0.05), that of the exhaustion control group significantly increased (P＜0.01).
Compared to the exhaustion control group, the testosterone level in serum of Mingmen moxibustion+exhaustion group was significantly decreased (P<0.01), but the change of Guanyuan moxibustion+exhaustion group was not significant (P>0.05), and there were significant differences between the two moxibustion+exhaustion groups (P<0.05).
10Ghrelin level in hypothalamus and serum
Compared to the normal control group, the ghrelin level in hypothalamus of Guanyuan moxibustion group and Mingmen moxibustion group had no significant changes (P>0.05), that of the exhaustion control group significantly increased (P<0.01).
Compared to the exhaustion control group, the ghrelin level in hypothalamus of Mingmen moxibustion+exhaustion group was significantly decreased (P<0.01), but the change of Guanyuan moxibustion+exhaustion group was not significant (P>0.05), and there were significant differences between the two moxibustion+exhaustion groups (P<0.05). The change pattern of ghrelin level in serum was the same as that in hypothalamus.
Conclusions:
1Preventive moxibustion was benign stress. Its regulation on the body was not be shown by the changes of biochemical index. But obviously, it enabled the body to perform better in the resistance to harmful stress.
2Exhaustive exercise was a harmful over-stressor that led to the excessive stress of the body. In the excessive stress, stress axis was over-activated, energy reserve was over-used and energy metabolism was in a state of disorder.
3Preventive moxibustion at both Guanyuan(RN-4) and Mingmen(DU-4) have a significant improvement on rat's exercise tolerance, which indicates its potential value in sport medicine. The beneficial regulation of preventive moxibustion possibly related to the balance of energy metabolism as a result of the adjustment of stress response, HPG axis, NOS activity and ghrelin level. The effect was accomplished by the increase of the comprehensive abilities of the body to keep the balance and stability in the internal environment. We called it the self-rebalance ability. We hold that self-rebalance ability is the core of health energy (Zheng Qi).
4The effect of preventive moxibustion can be diverted by the property of different acu-points. Although the effect on swimming exhausted time and energy metabolism of Guanyuan point had no difference to that of Mingmen point, its effect on some indices might be delayed. Or the regulating effect of Guanyuan point on energy metabolism though different pathway. The difference between two acu-points needs to be further researched.
我们前期实验观察到逆灸可以减轻随后的“疾病状态”大鼠(佐剂性关节炎、痛经模型)的发病率与疾病程度,也对“亚健康状态”雌性大鼠机体(更年期模型)有良性的调节作用。我们想进一步观察逆灸对“健康”机体的作用。结合逆灸的作用特点,我们需要一个能体现机体综合素质的指标,以全面概括逆灸的“良性预应激”作用。
运动能力是动物的基本能力之一,体现机体的生命力,需要各个系统的全面配合,是机体综合素质的体现。强大的运动能力可以说是机体整体健康程度的一种外在表现,也可以说是机体“正气充足”的体现。我们可以通过观察逆灸对动物运动耐力的影响,进而推测逆灸是如何通过“扶助”正常机体的“正气”而达到防病保健的效果。极限运动耐力属于运动能力的一种,可以使机体将自身潜力充分发挥出来,更能体现出机体的生命潜力。同时,极限耐力运动属于过度应激,与逆灸的适度应激相对应,两者一前一后可以体现出两种应激叠摞后的效应。
能量代谢是动物所有功能活动的基础,应激是机体内环境面对自身及外环境的变动而产生的变化,两者的稳定对维持整个机体的稳定具有重要意义。极限耐力运动需要持续的能量供应,同时也是一种会造成机体过度应激反应的不良应激原,因此极限耐力运动能够很好地将应激与能量代谢联系起来。Ghrelin由于其特殊的脑肠肽特性,可能在应激与能量代谢系统甚至其他系统之间的稳态调节都起到关键因子的作用。应激、能量代谢与Ghrelin水平三者可能互相影响,互相调节,共同维持着机体的稳态。我们认为这些指标能够反映机体的自稳调节能力,可以通过观察这些指标在极限耐力运动中的变化探讨逆灸对身体素质的改善作用。
关元穴和命门穴是传统保健穴,联系人体的元气,作用比较基础、广泛,并且在穴性方面比较有代表性(一阴一阳)。选用两穴进行比较,一方面可以体现逆灸的共同效应,一方面又能反映逆灸不同穴位的特异性。
目的：通过观察逆灸关元穴和逆灸命门穴对大鼠极限运动耐力的提升作用,探讨逆灸的扶助正气、增强身体素质的效果。通过观察逆灸、力竭和逆灸+力竭对大鼠肝糖原、血乳酸、心肌和血清NOS、 HPA轴应激激素、HPG轴相关激素、下丘脑和血清Ghrelin水平等指标的影响,探讨逆灸对力竭运动大鼠Ghrelin的调节作用及其与能量代谢的相关性,进而探究逆灸提高机体运动耐力的部分机制,进一步揭示逆灸防病保健的科学内涵,或为运动医学提供参考。
方法：将48只SD雄性大鼠分为：空白对照组、逆灸关元组、逆灸命门组、力竭对照组、逆灸关元+力竭组和逆灸命门+力竭组。逆灸关元组、逆灸命门组、逆灸关元+力竭组和逆灸命门+力竭组分别行关元穴和命门穴艾条灸法,隔天1次,每次10min,连续10次；空白对照组和力竭对照组与上述组同时人工抓取但不灸,如此连续10次。第20天,即最后一次艾灸24h后,力竭对照组、逆灸关元+力竭组和逆灸命门+力竭组混合后随机抽取进行温水力竭游泳并记录游泳至力竭的时间。力竭即刻与空白对照组、逆灸关元组和逆灸命门组一同处死取材。观察逆灸关元穴和命门穴对大鼠力竭游泳时间的影响。采用生化法检测各组大鼠肝糖原、血乳酸、心肌和血清NOS活性、HPA轴应激激素水平(下丘脑CRH、垂体ACTH血清CORT)、HPG轴相关激素(下丘脑GnRH、血清睾酮)、中枢及外周Ghrelin水平。
结果：
1力竭时间
逆灸关元+力竭组和逆灸命门+力竭组的游泳力竭平均时间都明显长于力竭对照组(P<0.01,P<0.05)；两个逆灸+力竭组之间无显著差异(P>0.05)。
2肝糖原含量
逆灸关元组和逆灸命门组的肝糖原含量与空白对照组相比没有显著变化(P>0.05)。力竭对照组的肝糖原含量较空白对照组极显著减少(P<0.01)。逆灸关元+力竭组与逆灸命门+力竭组的肝糖原较力竭对照组均有极显著或显著升高(P<0.01,P<0.05)。两个逆灸+力竭组之间无显著差异(P>0.05)。
3血乳酸含量
逆灸关元组和逆灸命门组的血乳酸含量与空白对照组相比均没有明显变化(P>0.05)。力竭对照组的血乳酸较空白对照组极显著升高(P<0.01)。逆灸关元+力竭组与逆灸命门+力竭组的血乳酸与力竭对照组相比均有极显著的降低(P<0.01)。两个逆灸+力竭组之间无差异(P>0.05)。
4心肌和血清NOS活性
与空白对照组相比,逆灸关元组和逆灸命门组的心肌NOS活性没有明显变化(P>0.05),力竭对照组的心肌NOS活性极显著升高(P<0.01)；与力竭对照组比较,逆灸关元+力竭组心肌NOS活性没有明显变化(P>0.05),逆灸命门+力竭组心肌NOS活性极显著下降(P<0.01)。两个逆灸+力竭组之间有显著统计学差异(P<0.05)。
各组血清NOS活性和心肌NOS活性的变化情况一致。
5下丘脑CRH含量
与空白对照组比较,逆灸关元穴组和逆灸命门穴组大鼠的下丘脑CRH含量均无明显变化(P>0.05)。力竭对照组大鼠的下丘脑CRH含量较空白对照组有极显著升高(P<0.01)。与力竭对照组比较,逆灸关元+力竭组大鼠的下丘脑CRH含量没有变化(P>0.05)；逆灸命门+力竭组大鼠的下丘脑CRH含量极显著降低(P<0.01)。两个逆灸+力竭组之间有极显著统计学差异(P<0.01)。
6垂体ACTH含量
与空白对照组比较,逆灸关元穴组和逆灸命门穴组大鼠的垂体ACTH含量均无明显变化(P>0.05)。力竭对照组大鼠的垂体ACTH含量较空白对照组有极显著升高(P<0.01)。与力竭对照组比较,逆灸关元+力竭组大鼠的垂体ACTH含量没有变化(P>0.05)；逆灸命门+力竭组大鼠的垂体ACTH含量极显著降低(P<0.01)。两个逆灸+力竭组之间有极显著统计学差异(P<0.01)。
7血清CORT含量
与空白对照组比较,逆灸关元穴组和逆灸命门穴组大鼠的血清CORT含量均无明显变化(P>0.05)。力竭对照组大鼠的血清CORT含量较空白对照组有极显著升高(P<0.01)。与力竭对照组比较,逆灸关元+力竭组大鼠的血清CORT含量没有变化(P>0.05)；逆灸命门+力竭组大鼠的血清CORT含量极显著降低(P<0.01)。两个逆灸+力竭组之间有极显著统计学差异(P<0.01)。
8下丘脑GnRH含量
与空白对照组相比,逆灸命门穴组大鼠的下丘脑GnRH含量无明显变化(P>0.05),力竭对照组大鼠的下丘脑GnRH极显著升高(P<0.01)。与力竭对照组比较,逆灸关元+力竭组大鼠下丘脑GnRH含量没有明显变化(P>0.05)；逆灸命门+力竭组大鼠下丘脑GnRH含量极显著降低(P<0.01)。两个逆灸+力竭组之间的下丘脑GnRH含量有显著统计学差异(P<0.05)。
9血清睾酮含量
与空白对照组相比,逆灸命门穴组大鼠的血清睾酮含量无明显变化(P>0.05),力竭对照组大鼠的血清睾酮极显著升高(P<0.01)。与力竭对照组比较,逆灸关元+力竭组大鼠血清睾酮含量没有明显变化(P>0.05)；逆灸命门+力竭组大鼠血清睾酮含量极显著降低(P<0.01)。两个逆灸+力竭组之间的血清睾酮含量差异有显著统计学意义(P<0.05)。
10下丘脑和血清Ghrelin含量
与空白对照组相比,逆灸命门穴组大鼠的下丘脑Ghrelin含量无明显变化(P>0.05),力竭对照组大鼠的下丘脑Ghrelin含量极显著升高(P<0.01)。与力竭对照组比较,逆灸关元+力竭组大鼠下丘脑Ghrelin含量没有明显变化(P>0.05)；逆灸命门+力竭组大鼠下丘脑Ghrelin含量极显著降低(P<0.01)。两个逆灸+力竭组之间的下丘脑Ghrelin含量有显著统计学差异(P<0.05)。
各组血清Ghrelin和下丘脑Ghrelin含量的变化情况一致。
结论：
1逆灸对机体是一种适度的应激,对机体的调节作用不会预先在单个生化指标上体现出来。但却能显著改善机体的健康程度,提高机体对不良应激的抵抗力。
2力竭运动是一种能够引起机体过度应激反应的不良应激原。力竭导致机体应激轴过度激活,过分调动了能源储备,能量代谢处于紊乱状态。
3我们认为“正气”的核心是机体的“自稳调节能力”。逆灸正是通过提高机体的自稳调节能力以扶助正气,进而增强机体素质,提高运动耐力。这可能能够为运动医学提供一定的参考。其具体机制可能与调节应激激素,能量代谢相关因子和Ghrelin水平等有关。
4关元穴与命门穴的效应有所不同。关元穴在延长大鼠游泳时间、调节糖原储备和乳酸代谢方面与命门穴无异,但对应激激素、性激素、NOS活性及Ghrelin水平的调节效应不明显,关元穴对这些指标的效应可能具有滞后性,也可能其对大鼠体能的提升作用以及对能量代谢的调节作用是通过其他途径实现的。两穴差别有待我们进一步研究探讨。
Preventive acupuncture and moxibustion are important methods of treating pre-disease in traditional Chinese medicine, which have played an important role in disease prevention, treatment, health care and life extension. According to recent researches, the regulation effect of preventive mexibustion is extensive and involving many fundamental physiological processes. The features of preventive moxibustion can be summarized into three traits:beneficial, extensive, and latent. According to the theory of traditional Chinese medicine, preventive moxibustion enhance the anti-disease ability by building up the health energy (Zheng Qi). We hold that self-rebalance ability is the core part of health energy (Zheng Qi). As a benign stressor giving mild pre-stress to the body, moxibustion will enhance the self-rebalance ability and correct the potential dysfunctions to enable the body to mobilize more potential abilities to resist the damages and illnesses causing by the following over-stress. In our latest experiment, we observed that preventive moxibustion could reduce incidence and severity of disease (adjuvant arthritis, dysmenorrhea model) and give benign regulation to "sub-health" state (menopausal model) in modeled rats. Therefore, we are going to observe the effect of preventive moxibustion on healthy state of animals. Considering the complexity of the comprehensive effect, we need a physiological index that can be representative for the comprehensive quality of the body and the "benign pre-stress" effect of preventive moxibustion. Locomotivity is one of the basic abilities representing the vitality of the body. According to physiology studies, locomotivity depend on the ensemble coordinating operation of all the organs and systems of the body. According to the theory of traditional Chinese medicine, strong locomotivity means strong health energy (Zheng Qi), which is the comprehensive representation of the health condition of the human body. Through observing the effect of preventive moxibustion on locomotivity of animals, we can know how the body will be healthier by strengthening the health energy (Zheng Qi) with preventive moxibustion. As a kind of locomotivity, extreme exercise endurance, being able to force the body to fully perform its potential, is representative of the comprehensive quality of the body. Meanwhile, being a harmful over-stressor that will lead to excessive stress, extreme exercise endurance can be paired with preventive moxibustion in the comparison of the effect of benign pre-stress and harmful over-stress.
Energy metabolism is fundamental for all the functional activity of animal; and stress is the change of internal environment to external environment. The stability of energy metabolism and stress is important in the maintaining of the stability of the whole body. Being a harmful stressor, extreme exercise will not only cause over-stress response but also needs continuous supply of energy, therefore it can be a link between stress and energy metabolism. As a brain-gut peptide, ghrelin may be playing a key role in regulating the balance between energy metabolism and stress response or even other systems. Stress, energy metabolism and ghrelin level may have influence on each other in the keeping of the homeostasis. We hold that these index can represent the self-rebalance ability of the body, and their changes in exhaustive exercise can be useful in the study of effect of preventive moxibustion on the health of the body. Guanyuan(RN-4) and Mingmen(DU-4)point point are the most commonly used points in health keeping because of their extensive effects and their opposite nature in Yin-Yang. Comparing the two point in the experiment not only can represent the common effect of moxibustion but also show the deviation effect of different acu-points.
Objective:To discuss the enhancement of health energy and improvement of constitution with preventive moxibustion through observing the effect of preventive moxibustion at Guanyuan and Mingmen points on exercise endurance of rats. To discuss the correlation and influence of preventive moxibustion on ghrelin level and energy metabolism of exhaustive swimming rats through observation of the changes of liver glycogen, serum lactate, myocardium NOS, serum NOS, hormone of HPA axis, hormone of HPG axis, hypothalamus ghrelin and serum ghrelin before and after preventive moxibustion and exhausted swimming. And also introduce preventive moxibustion to sports medicine.
Methods:Forty-eight male SD rats were divided into six groups:normal control group, Guanyuan moxibustion group, Mingmen moxibustion group, exhaustion control group, Guanyuan moxibustion+exhaustion group and Mingmen moxibustion+exhaustion group. All moxibustion groups received a ten-minute moxibustion every two days, ten times in all. The two control groups received sham treatment at the same time with moxibustion groups. In the twentith day, twenty four hours after the last moxibustion, exhaustion control group, Mingmen moxibustion+exhaustion group and Guanyuan moxibustion+exhaustion group received a warm water exhaustive swimming and the swimming exhausted time was recorded. After swimming exhaustion, all rats were executed and sampled whole blood, right lobe of liver, heart muscle, hypothalamus, hypophysis and right side adrenal gland. The swimming time of all swimming groups were compared. Glycogen level in liver, lactate level in serum, NOS level in myocardium, NOS level in serum, hormone level of HPA axis, hormone level of HPG axis, ghrelin level in hypothalamus and ghrelin level in serum were detected.
Results:
1Swimming exhausted time
Compared to the exhaustion control group, the swimming exhausted time of Guanyuan moxibustion+exhaustion group and Mingmen moxibustion+exhaustion group were significantly higher(P<0.01, P＜0.05), there was no difference between the two moxibustion+exhaustion groups (P>0.05)
2Glycogen level in liver
Compared to the normal control group, the glycogen level in liver of Guanyuan moxibustion group and Mingmen moxibustion group had no significant changes (P>0.05), that of the exhaustion control group significantly decreased (P＜0.01).
Compared to the exhaustion control group, the glycogen level in liver of Guanyuan moxibustion+exhaustion group and Mingmen moxibustion+exhaustion group were significantly increased (P＜0.01, P<0.05), and there was no difference between the two moxibustion+exhaustion groups (P>0.05).
3lactate level in blood
Compared to the normal control group, the lactate level in blood of Guanyuan moxibustion group and Mingmen moxibustion group had no significant changes (P>0.05), that of the exhaustion control group significantly increased (P<0.01).
Compared to the exhaustion control group, the lactate level in blood of Guanyuan moxibustion+exhaustion group and Mingmen moxibustion+exhaustion group were significantly dropped (P＜0.01), and there was no difference between the two moxibustion+exhaustion groups (P>0.05).
4NOS level in myocardium and serum
Compared to the normal control group, the NOS level in myocardium of Guanyuan moxibustion group and Mingmen moxibustion group had no significant changes (P>0.05), that of the exhaustion control group significantly increased (P＜0.01).
Compared to the exhaustion control group, the NOS level in myocardium of Mingmen moxibustion+exhaustion group was significantly decreased (P<0.01), but the change of Guanyuan moxibustion+exhaustion group was not significant (P>0.05), and there were significant differences between the two moxibustion+exhaustion groups (P＜0.05). The change pattern of NOS level in serum was the same as that in myocardium.
5CRH level in hypothalamus
Compared to the normal control group, the CRH level in hypothalamus of Guanyuan moxibustion group and Mingmen moxibustion group had no significant changes (P>0.05), that of the exhaustion control group significantly increased (P<0.01).
Compared to the exhaustion control group, the CRH level in hypothalamus of Mingmen moxibustion+exhaustion group was significantly decreased (P<0.01), but the change of Guanyuan moxibustion+exhaustion group was not significant (P>0.05), and there were significant differences between the two moxibustion+exhaustion groups (P<0.05).
6ACTH level in hypophysis
Compared to the normal control group, the ACTH level in hypophysis of Guanyuan moxibustion group and Mingmen moxibustion group had no significant changes (P>0.05), that of the exhaustion control group significantly increased (P<0.01).
Compared to the exhaustion control group, the ACTH level in hypophysis of Mingmen moxibustion+exhaustion group was significantly decreased (P<0.01), but the change of Guanyuan moxibustion+exhaustion group was not significant (P>0.05), and there were significant differences between the two moxibustion+exhaustion groups (P＜0.05).7Cortisol level in serum
Compared to the normal control group, the cortisol level in serum of Guanyuan moxibustion group and Mingmen moxibustion group had no significant changes (P>0.05), that of the exhaustion control group significantly increased (P<0.01).
Compared to the exhaustion control group, the cortisol level in serum of Mingmen moxibustion+exhaustion group was significantly decreased (P＜0.01), but the change of Guanyuan moxibustion+exhaustion group was not significant (P>0.05), and there were significant differences between the two moxibustion+exhaustion groups (P<0.01).
8GnRH level in hypothalamus
Compared to the normal control group, the GnRH level in hypothalamus of Guanyuan moxibustion group and Mingmen moxibustion group had no significant changes (P>0.05), that of the exhaustion control group significantly increased (P<0.01).
Compared to the exhaustion control group, the GnRH level in hypothalamus of Mingmen moxibustion+exhaustion group was significantly decreased (P<0.01), but the change of Guanyuan moxibustion+exhaustion group was not significant (P>0.05), and there were significant differences between the two moxibustion+exhaustion groups (P＜0.05).
9Testosterone level in serum
Compared to the normal control group, the testosterone level in serum of Guanyuan moxibustion group and Mingmen moxibustion group had no significant changes (P>0.05), that of the exhaustion control group significantly increased (P＜0.01).
Compared to the exhaustion control group, the testosterone level in serum of Mingmen moxibustion+exhaustion group was significantly decreased (P<0.01), but the change of Guanyuan moxibustion+exhaustion group was not significant (P>0.05), and there were significant differences between the two moxibustion+exhaustion groups (P<0.05).
10Ghrelin level in hypothalamus and serum
Compared to the normal control group, the ghrelin level in hypothalamus of Guanyuan moxibustion group and Mingmen moxibustion group had no significant changes (P>0.05), that of the exhaustion control group significantly increased (P<0.01).
Compared to the exhaustion control group, the ghrelin level in hypothalamus of Mingmen moxibustion+exhaustion group was significantly decreased (P<0.01), but the change of Guanyuan moxibustion+exhaustion group was not significant (P>0.05), and there were significant differences between the two moxibustion+exhaustion groups (P<0.05). The change pattern of ghrelin level in serum was the same as that in hypothalamus.
Conclusions:
1Preventive moxibustion was benign stress. Its regulation on the body was not be shown by the changes of biochemical index. But obviously, it enabled the body to perform better in the resistance to harmful stress.
2Exhaustive exercise was a harmful over-stressor that led to the excessive stress of the body. In the excessive stress, stress axis was over-activated, energy reserve was over-used and energy metabolism was in a state of disorder.
3Preventive moxibustion at both Guanyuan(RN-4) and Mingmen(DU-4) have a significant improvement on rat's exercise tolerance, which indicates its potential value in sport medicine. The beneficial regulation of preventive moxibustion possibly related to the balance of energy metabolism as a result of the adjustment of stress response, HPG axis, NOS activity and ghrelin level. The effect was accomplished by the increase of the comprehensive abilities of the body to keep the balance and stability in the internal environment. We called it the self-rebalance ability. We hold that self-rebalance ability is the core of health energy (Zheng Qi).
4The effect of preventive moxibustion can be diverted by the property of different acu-points. Although the effect on swimming exhausted time and energy metabolism of Guanyuan point had no difference to that of Mingmen point, its effect on some indices might be delayed. Or the regulating effect of Guanyuan point on energy metabolism though different pathway. The difference between two acu-points needs to be further researched.
引文
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