活血荣络方对脑缺血再灌注损伤大鼠SIRT1/PGC-1α信号通路的影响

Effects of Huoxue Rongluo Decoction on Rats with Cerebral Ischemia-reperfusion Injury Based on SIRT1/PGC-1αSignaling Pathway

目的观察活血荣络方对脑缺血再灌注损伤(CIRI)大鼠SIRT1/PGC-1α信号通路的影响,初步探讨其对CIRI的保护机制。方法60只大鼠随机分为假手术组、模型组、活血荣络方组、EX-527组、EX-527+活血荣络方组和艾地苯醌组,采用线栓法建立CIRI大鼠模型。给药组于造模前36 h首次予相应药物灌胃(10 mL/kg),之后每24 h给药1次,共3次,EX-527组和活血荣络方+EX-527组于再灌注前20 min腹腔注射EX-527(5 mL/kg),其余各组在相同时间灌胃蒸馏水或腹腔注射生理盐水。再灌注24 h后对大鼠进行神经功能缺损评分,HE染色观察缺血皮质区损伤情况,试剂盒检测皮质区三磷酸腺苷(ATP)含量,免疫组化检测缺血皮质区沉默信息调节因子1(SIRT1)、过氧化物酶体增殖物激活受体γ辅激活因子1α(PGC-1α)蛋白表达,RT-qPCR检测缺血皮质区SIRT1、PGC-1αmRNA表达。结果与假手术组比较,模型组大鼠神经功能缺损评分升高,缺血皮质区神经元损伤严重,ATP含量减少,SIRT1、PGC-1α蛋白和mRNA表达降低(P<0.05)。与模型组比较,活血荣络方组和艾地苯醌组大鼠神经功能缺损评分降低,缺血皮质区神经元损伤改善,ATP含量增加,SIRT1、PGC1-α蛋白和mRNA表达升高(P<0.05);EX-527组大鼠神经功能缺损评分升高,缺血皮质区神经元损伤加重,ATP含量减少,SIRT1、PGC-1α蛋白和mRNA表达降低(P<0.05)。与活血荣络方组比较,活血荣络方+EX-527组大鼠神经功能缺损评分升高,缺血皮质区神经元坏死和凋亡增多,ATP含量减少,SIRT1、PGC-1α蛋白和mRNA表达降低(P<0.05)。结论活血荣络方可促进CIRI大鼠ATP生成,改善神经功能缺损症状,其机制可能与激活SIRT1/PGC-1α信号通路有关。

Objective To observe the effects of Huoxue Rongluo Decoction on SIRT1/PGC-1αsignaling pathway in rats with cerebral ischemia-reperfusion injury(CIRI);To conduct preliminary study on its mechanism of intervention on CIRI.Methods Sixty SD rats were randomly divided into sham-operation group,model group,Huoxue Rongluo Decoction group,EX-527 group,EX-527+Huoxue Rongluo Decoction group and idebenone group.The CIRI rat model was established by thread occlusion.The corresponding drugs were given by intragastric administration(10 mL/kg)to medicine groups 36 h before the establishment of the model,and then every 24 hours for 3 times,the EX-527 group and EX-527+Huoxue Rongluo Decoction group were given EX-527(5 mL/kg)by intraperitoneal injection 20 minutes before reperfusion by gavage or injected intraperitoneallywith normal saline,the other groups were given distilled water at the same time.The rats were scored for neurological deficits 24 h after reperfusion,the injury of ischemic cortex was observed by HE staining,the content of ATP in ischemic cortex was detected by kit,the expressions of SIRT1 and PGC-1αprotein in ischemic cortex were detected by immunohistochemistry,the expressions of SIRT1 and PGC-1αmRNA in ischemic cortex were detected by RT-qPCR.Results Compared with the sham-operation group,the neurological deficit score of rats in the model group increased significantly,the neuron injury in ischemic cortex was serious,the content of ATP reduced,and the expressions of SIRT1,PGC-1αprotein and mRNA decreased(P<0.05).Compared with the model group,the neurological deficit score of rats in the Huoxue Rongluo Decoction group and the idebenone group decreased,the neuronal injury in ischemic cortex improved,the content of ATP increased,and the expressions of SIRT1,PGC-1αprotein and mRNA increased(P<0.05);The neurological deficit score in the EX-527 group increased,the neuronal injury in ischemic cortex aggravated,the content of ATP reduced,the expressions of SIRT1,PGC-1αprotein and mRNA decreased(P<0.05).Compared with Huoxue Rongluo Decoction group,the neurological deficit score in Huoxue Rongluo Decoction+EX-527 group increased,the neuronal necrosis and apoptosis in ischemic cortex increased,the content of ATP reduced,the expression of SIRT1,PGC-1αprotein and mRNA decreased(P<0.05).Conclusion Huoxue Rongluo Decoction can promote the production of ATP in rats with CIRI,improve the symptoms of neurological deficit,and the mechanism may be related to the activation of SIRT/PGC-1αsignaling pathway.