灵龟八法艾灸对衰老大鼠肝脏端粒长度及细胞周期调节因子表达的影响

Effect of moxibustion on acupoints at “opening” time on telomere length and expression of liver cell cycle regulators in aging rats

目的:观察灵龟八法艾灸对衰老大鼠肝脏中端粒长度和抑癌基因p53及视网膜母细胞瘤基因(Rb)表达的影响,探讨灵龟八法艾灸延缓衰老的作用机制。方法:将40只SD大鼠随机分为正常组、模型组、预防组和治疗组,每组10只。除正常组外,其余3组大鼠用腹腔注射D-半乳糖溶液(200 mg/kg)的方法制备衰老模型,每天1次,共42 d。预防组在造模期间同时给予灵龟八法按时开穴灸,造模结束后继续常规喂养28 d;治疗组在造模期间不做治疗,在造模结束的次日进行灵龟八法按时开穴灸,共28 d。用实时荧光定量PCR法检测肝脏中端粒相对长度及p53和Rb的mRNA表达;用酶联免疫吸附法检测肝脏中p53和Rb的蛋白含量。结果:与正常组比较,模型组大鼠端粒相对长度明显缩短(P<0.01),p53和Rb的mRNA表达和蛋白含量均明显增加(P<0.01)。与模型组比较,预防组和治疗组的端粒相对长度均显著增加(P<0.01), p53和Rb mRNA表达和蛋白含量均显著降低(P<0.01,P<0.05)。灵龟八法预防组和灵龟八法治疗组组间各指标比较差异无统计学意义(P>0.05)。结论:灵龟八法按时开穴灸能抑制衰老大鼠肝脏端粒长度的缩短,并下调p53、Rb的mRNA表达和蛋白含量,延缓细胞衰老的进程。

Objective To observe the effects of "lingguibafa" moxibustion performing at the appropriate acupoints at their "opening" time on telomere length,expressions of p53 of tumor supressor genes and retinoblastoma gene(Rb)in the liver of aging rats,so as to explore its mechanisms underlying delaying senescence. Methods Forty male SD rats were randomly divi-ded into normal, model, prevention and treatment groups, with 10 rats in each group. The rat model was established by intrape-ritoneally injection of D-galactose(200 mg/kg) once a day for 42 days. The rats in the prevention group were given "lingguibafa" moxibustion during modeling, while those in the treatment group were given "lingguibafa" moxibustion after modeling. Quantitative real-time PCR was used to detect the telomere length and the mRNA expressions of p53 and Rb,ELISA was used to detect the protein contents of p53 and Rb in the liver tissues. Results Compared with the normal group, the relative telomere length of the model group was significantly shortened(P<0.01), the mRNA expressions and protein contents of p53 and Rb were significantly increased(P<0.01). After intervention and in comparison with the model group, the relative telomere length of the prevention group and the treatments group were significantly increased(P<0.01), and the expressions of p53 and Rb mRNA and protein contents were significantly reduced(P<0.01, P<0.05). There were no significant difference between the prevention and the treatment groups in the abovementioned indexes(P>0.05). Conclusion Moxibustion on acupoints at "opening" time can inhibit the shortening of telomere length and the down-regulation of the expressions of p53 and Rb in aging rats, which may contribute to its function in delaying the process of cell senescence.