针刺对SAMP8小鼠海马线粒体SIRT3/PGC-1α蛋白表达的影响

Effect of acupuncture on SIRT3/PGC-1αprotein expression in hippocampus of SAMP8 mice

目的观察针刺百会穴、涌泉穴对阿尔兹海默病小鼠学习记忆功能、海马区线粒体SIRT3、PGC-1α蛋白表达的影响。方法选取6月龄SAMP8小鼠,随机分为模型组、治疗组、抑制剂组,以SAMR1小鼠作为空白组。治疗组、抑制剂组均予以百会、涌泉穴针刺治疗,每日1次,5日1疗程,共4个疗程;针刺结束后行3-TYP、Morris水迷宫干预处理,行为学检测评估空间学习记忆能力,RT-PCR、免疫荧光、蛋白免疫印迹检测海马SIRT3、PGC-1α的mRNA及蛋白表达。结果与空白组比较,模型组的逃避潜伏期在第4、5、7天明显延长(P<0.05,P<0.01);在第4、7天,治疗组的逃避潜伏期较模型组明显降低(P<0.05);与治疗组相比,抑制剂组第6天的潜伏期明显延长,结果具有统计学意义(P<0.05);与空白组比较,模型组小鼠的跨平台次数、目标象限的游泳时间/总时间比、目标象限的游泳路程/总路程比均显著减少(P<0.01),与模型组比较,治疗组小鼠均显著增加(P<0.01);抑制剂组与治疗组结果无统计学差异(P>0.05)。与空白组比较,模型组SIRT3、PGC-1α的mRNA相对表达含量显著增加,有统计学差异(P<0.01,P<0.05);与模型组比较,治疗组的表达呈下调趋势(P<0.01,P<0.05),且抑制剂组和治疗组之间无统计学差异(P>0.05)。与空白组比较,模型组小鼠海马区SIRT3、PGC-1α的表达均明显减少(P<0.01);与模型组相比,治疗组的蛋白阳性表达均明显增加(P<0.01);与治疗组比较,抑制剂组均显著减少(P<0.01)。结论针刺对SAMP8小鼠学习记忆能的提升,可能与针刺调控SIRT3、PGC-1α蛋白表达,保护能量代谢过程,改善线粒体功能障碍有关。

Objective To observe the effect of acupuncture at Baihui and Yongquan on the learning and memory function,the expression of SIRT3 and PGC-1αprotein in the hippocampus of Alzheimer’s disease(AD)mice.Methods SAMP8 mice of 6 month old were randomly divided into model group,treatment group and inhibitor group,and SAMR1 mice as the blank group.The treatment group and the inhibitor group were treated with Baihui and Yongquan points acupuncture once a day,five days a course,a total of four courses;after the acupuncture,3-typ and Morris water maze intervention were conducted,and the spatial learning and memory ability was assessed by behavioral test,and the mRNA and protein expression of SIRT3 and PGC-1αin hippocampus were detected by RT-PCR,immunofluorescence and Western blotting.Results compared with the blank group,the escape latency of the model group was significantly longer on the 4 th,5 th and 7 th day(P<0.05,P<0.01);on the 4 th and 7 th day,the escape latency of the treatment group was significantly lower than that of the model group(P<0.05);compared with the treatment group,the latency of the inhibitor group on the 6 th day was significantly longer,and the results were statistically significant(P<0.05);compared with the blank group,the model group mice had a cross leveling Compared with the model group,the mice in the treatment group were significantly increased(P<0.01);the results of the inhibitor group and the treatment group were not statistically different(P>0.05).Compared with the blank group,the relative expression of SIRT3 and PGC-1αmRNA in the model group increased significantly(P<0.01,P<0.05);compared with the model group,the expression in the treatment group decreased(P<0.01,P<0.05),and there was no statistical difference between the inhibitor group and the treatment group(P>0.05).Compared with the blank group,the expression of SIRT3 and PGC-1αin the hippocampus of the model group was significantly reduced(P<0.01);compared with the model group,the protein positive expression in the treatment group was significantly increased(P<0.01);compared with the treatment group,the inhibitor group was significantly reduced(P<0.01).Conclusion the improvement of learning and memory ability of SAMP8 mice by acupuncture may be related to the regulation of SIRT3 and PGC-1αprotein expression by acupuncture,the protection of energy metabolism and the improvement of mitochondrial dysfunction.