不同取穴方法对亚急性衰老大鼠的影响及其作用机制的比较研究
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摘要
目的:研究针刺防治衰老的作用及其机理,通过对三组抗衰老穴位防治衰老作用机制的比较,旨初步观察抗衰老穴位是否存在特异性,穴位间是否存在拮抗及协同作用,为针灸抗衰老在穴位的选择和配伍上提供理论支持和实验依据。
     方法:采用健康Wistar大鼠作为受试对象,随机分为五组,即正常组、模型组、督穴组、体穴组、督体穴组。以D-半乳糖致亚急性衰老大鼠作为模型组,正常大鼠作对照,观察大鼠的一般状态及脑组织形态学改变,通过对自由基的影响、抗氧化防御能力、细胞膜离子转运活性及脑、肝组织蛋白表达等方面的检测,对针刺防治衰老的部分作用机制进行探讨,并对三组抗衰老穴位防治衰老的作用及其机理进行横向对比研究。
     结果:
     1.针刺对血清中抗活性氧能力和抗超氧阴离子自由基活力的下降,及对血清、肝组织中H_2O_2含量的升高均有一定的抑制作用,具有督体穴组作用较强,督穴组、体穴组差异不大的趋势及特点,这可能与针刺疗程长短有一定的关系;
     2.各针刺组抗脂质过氧化作用均比较全面,总体而言,在延缓T-AOC、SOD、CAT和GSH-PX含量降低,抑制MDA、NO和NOS含量升高方面督体穴组作用最强;对于改善脑组织抗氧化能力,督穴组作用较体穴组好;
     3.各针刺组均有不同程度延缓衰老大鼠脑、肝细胞膜Na~+-K~+-ATPase及Ca~(2+)-Mg~(2+)-ATPase活性下降的作用,其中督体穴组作用最强:对于改善脑细胞膜离子转运能力,体穴组作用不及督穴组;
     4.各针刺组均有抑制脑、肝组织Bcl-2蛋白表达下降及Bax蛋白过度表达,从而抑制Bcl-2/Bax比值下降的作用,同时各针刺组还具有抑制脑、肝组织Caspase-3、p16蛋白表达升高的作用,体现出督体穴组作用最强;对脑蛋白表达的影响,督穴组较体穴组作用好;
     5.各针刺组均可延缓脑组织细胞形态的异常变化,督体穴组作用最好,督穴组次之。
     结论:
     1.针刺具有显著预防衰老的作用,其可能通过减少机体内自由基、提高机体抗氧化防御能力、增强细胞膜离子转运活性、抑制细胞凋亡和延缓因衰老所致的细胞形态学改变等方面实现的。
     2.针刺防治衰老的作用以督体穴组表现的最为全面和稳定,在改善脑组织衰老方面督穴组体现出相应的特异性,提示了抗衰老穴位间具有协同性和特异性的特点。
     3.针刺对机体内自由基浓度的升高具有一定的抑制作用,同时可增强机体的抗氧化能力,其作用较为全面,表现为延缓T-AOC、SOD、CAT和GSH-PX活性的降低,抑制MDA、NO和NOS含量的升高,从而提示,针刺防治衰老的作用可能与其通过多途径减少机体内自由基含量及抑制自由基的脂质过氧化反应相关。
     4.针刺具有延缓脑、肝细胞膜Na~+-K~+-ATPase及Ca~(2+)-Mg~(2+)-ATPase活性下降的作用,由此提示其具有减少自由基对细胞膜的损伤,维持细胞膜稳定性的作用,这可能是其发挥抗衰老作用的机制之一。
     5.针刺具有抑制脑、肝组织Bcl-2蛋白表达和Bcl-2/Bax比值下降的作用,还具有抑制Bax、Caspase-3和p16蛋白过度表达的作用,从而说明针刺防治衰老作用的机理之一可能与其对细胞凋亡具有调控作用相关。
     6.针刺可延缓衰老大鼠大脑组织学超微结构的变化,通过维持细胞形态的正常,以确保其各项功能的发挥,从而起到预防衰老的作用,这可能是其作用机制之一。
Objective:To study on the effect and its mechanism of anti-aging treatedby acupuncture. Through comparing the mechanism of aging prevention by 3groups of anti-aging points, to observe if there is specificity in anti-agingpoints selection, and whether or not there is antagonism and synergy amongthe points, and to provide theoretical support and experimental basis for theselection and compatibility Of points for anti-aging treated by acupuncture.
     Methods:Choose healthy wistar rats as test objects and divided them into5 groups at randomly which include normal group, model group, dumeridian(DU) point group, body point group, DU and body point group. Usesubacute aging rats that are caused by D-galactose as model group, normalrats as control. Observe general condition of the rats and changes ofhistomorphology of their brain. Through detection of the influence on freeradicals, anti-oxidation capability, the activity of ion transportation ofmembrane, and the expression of tissue protein in brain and liver, toinvestigate the mechanism of anti-aging by acupuncture, and makecomparative research on the effect and its mechanism of 3 groups anti-agingpoints for anti-aging.
     Results:
     1.Acupuncture has certain inhibit effect on the ability of anti-activityoxygen in blood serum and decrease the activity of super anion free radical,and increase the content of H_2O_2 in blood serum arid hepatic tissue. It hasthe feature that the effect of DU and body point group is stronger, and thereis not big difference between DU point group and body point group. Thatmay have some relationship with the length of treatment course.
     2.The function of anti-lipid peroxidation of every acupuncture group isrelatively comprehensive. In all, on the aspects of postponing the decreasingof the content of T-AOC、SOD、CAT and GSH-PX, inhibiting the increasingof the content of MDA、NO and NOS, DU and body point group have the strongest effect. The DU point group is better than body point group onimproving the ability of antioxidation within brain tissue.
     3.Every acupuncture group have the effect of postponing the decreasingof the activity of Na~+-K~+-ATPase and Ca~(2+)-Mg~(2+)-ATPase in brain andhepatocyte membrane of aging rats in different degree, and among all groupsthe DU and body point group have the strongest effect. DU point group isbetter than body point group in improving ion transport capacity ofamelioration.
     4.Every acupuncture group has the effect to inhibit the decline ofexpression of Bcl-2 albumen in brain and hepatic tissue and inhibit theexcessive expression of Bax albumen thus to inhibit the ratio ofBcl-2/Bax.Every acupuncture group also has the effect of inhibiting theincreasing of Caspase-3、p16 albumen. DU and body point group have thestrongest effect. DU point group is better than body point group ininfluencing on the expression of cerebral protein in brain albumen.
     5.Every acupuncture group has good effect of maintenance ofmorphology of cells in brain tissue, and the DU point group is better, andDU and body point group is the best.
     Conclusions:
     1.Acupuncture has significant effect on prevent aging. It may reducefree radical of body, improve the ability of antioxidation, enhance activity oftransportation of ion through membrane, and inhibit apoptosis and delaychanges of morphology which have been leaded by aging.
     2.DU and body point group has overall and stable effect on preventingand treating aging among those acupuncture methods. DU point group showsrelative specific features on improvement of aging in brain tissue. It cuesthat there is cooperativity and specificity in different anti-aging pointsgroups.
     3.Acupuncture has some inhibition effect on increasing concentration offree radical of body, at the same time, it can enhance ability of antioxidation, and its effect is overall. It shows that acupuncture can postpone thedecreasing of the content of T-AOC、SOD、CAT and GSH-PX, inhibit theincreasing of the content of MDA、NO and NOS. So it shows that the effectof anti-aging by acupuncture may relate With the reduction of the content offree radical through many ways and the inhibition of lipid peroxidation.
     4.Acupuncture has the effect of postponing the decreasing of theactivity of Na~+-K~+-ATPase and Ca~(2+)-Mg~(2+)-ATPase in brain and hepatocytemembrane, so that it can reduce the damage of membrane by free radical, andmaintain the stability of membrane,which may be one of the mechanisms ofanti-aging by acupuncture.
     5.Acupuncture has the effect to inhibit the expression of Bcl-2 albumenof brain and hepatic tissue and descent the ratio of Bcl-2/Bax, and also hasthe effect to inhibit excessive expression of Bax, Caspase-3 and p16 albumen.So it shows that one of the mechanisms of prevention and curing aging byacupuncture may have correlate with the regulation to apoptosis.
     6. Acupuncture can delay the changes of histology ultrastructure ofcerebral tissue of aging rats, through maintain the normal forms of cells, toprevent aging. That may be one of the mechanisms.
引文
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