通督调神法针刺联合石冰醒脑颗粒对血管性痴呆大鼠肠道菌群微生物的影响

Effect of Tongdu Tiaoshen Acupuncture Combined with Shibing Xingnao Granule on Gut Microbiota in Rats with Vascular Dementia

目的探讨通督调神法针刺联合石冰醒脑颗粒对血管性痴呆大鼠肠道菌群的影响及其可能的作用机制。方法将80只大鼠随机分为假手术组、模型组、通督调神法针刺组、石冰醒脑颗粒组、通督调神法针刺联合石冰醒脑颗粒组,每组10只。采用改良双侧颈总动脉永久结扎法制备血管性痴呆模型。石冰醒脑颗粒组采用石冰醒脑颗粒溶解液灌胃,通督调神法针刺组采用通督调神针刺治疗,通督调神法针刺联合石冰醒脑颗粒组采用石冰醒脑颗粒溶解液联合通督调神法针刺治疗。采用Morris水迷宫实验评价各组大鼠认知和记忆功能变化,16S rRNA测序技术分析肠道菌群结构变化。Western blot法检测caspase-3、Bax和Bcl-2蛋白表达水平。结果与假手术组比较,模型组大鼠平均逃避潜伏期显著延长(P<0.05),caspase-3和Bax蛋白表达水平显著升高(P<0.05),Bcl-2蛋白表达水平显著降低(P<0.05);肠道中罗姆布茨菌等有益菌相对丰度显著降低(P<0.05)。与模型组比较,通督调神法针刺组、石冰醒脑颗粒组、通督调神法针刺联合石冰醒脑颗粒组大鼠平均逃避潜伏期显著缩短(P<0.05);caspase-3、Bax蛋白表达水平均显著降低(P<0.05)、Bcl-2蛋白表达水平均显著升高(P<0.05),肠道中密螺旋菌等有害菌相对丰度降低(P<0.05)。结论通督调神法针刺联合石冰醒脑颗粒可改善血管性痴呆大鼠认知功能障碍,其作用可能是通过调节海马caspase-3、Bax和Bcl-2蛋白表达,进一步影响肠道菌群而实现的。

Objective To investigate the effect of Tongdu Tiaoshen acupuncture combined with Shibing Xingnao Granule on gut microbiota in rats with vascular dementia and its possible mechanism of action.Methods A total of 80 rats were randomly divided into sham-operation group,model group,Tongdu Tiaoshen acupuncture group,Shibing Xingnao Granule group,and Tongdu Tiaoshen acupuncture+Shibing Xingnao Granule group,with 10 rats in each group.The modified method of permanent ligation of both common carotid arteries was used to establish a model of vascular dementia.The rats in the Shibing Xingnao Granule group were given the solution of Shibing Xingnao Granule by gavage,those in the Tongdu Tiaoshen acupuncture group were given Tongdu Tiaoshen acupuncture,and those in the Tongdu Tiaoshen acupuncture+Shibing Xingnao Granule group were given the solution of Tongdu Tiaoshen Acupuncture combined with Shibing Xingnao Granule.The Morris water maze test was used to evaluate the changes in cognitive and memory functions,and 16S rRNA sequencing was used to analyze the change in gut microbiota.Western blot was used to measure the protein expression levels of caspase-3,Bcl-2,and Bax.Results Compared with the sham-operation group,the model group had significant increases in mean escape latency(P<0.05)and the protein expression levels of caspase-3 and Bax(P<0.05),a significant reduction in the protein expression level of Bcl-2(P<0.05),and a significant decrease in the relative abundance of probiotic bacteria in the intestine(P<0.05).Compared with the model group,the Tongdu Tiaoshen acupuncture group,the Shibing Xingnao Granule group,and the Tongdu Tiaoshen acupuncture+Shibing Xingnao Granule group had significant reductions in mean escape latency(P<0.05),the protein expression levels of caspase-3,Bax(P<0.05),and the relative abundance of harmful intestinal bacteria(P<0.05),a significant increase in the protein expression level of Bcl-2(P<0.05).Conclusion Tongdu Tiaoshen acupuncture combined with Shibing Xingnao Granule can improve cognitive function in rats with vascular dementia,possibly by regulating the protein expression of caspase-3,Bax,and Bcl-2 in the hippocampus and affecting gut microbiota.