滋肾活血方对血管性痴呆大鼠分裂与融合蛋白Mfn1、Mfn2、Drp1、Fis1的影响

Effects of Zishen Huoxue Formula on fission and fusion proteins of Mfn1,Mfn2,Drp1 and Fis1 in vascular dementia rats

目的观察滋肾活血方对血管性痴呆(vascular dementia,VD)大鼠海马组织线粒体融合蛋白1(mitofusin 1,Mfn1)、线粒体融合蛋白2(mitofusin 2,Mfn2)、线粒体动力蛋白相关蛋白1(dynamin-related protein 1,Drp1)、线粒体分裂蛋白1(fission mitochondrial 1,Fis1)表达的影响。方法选用雄性SD大鼠60只,随机均分为假手术组、模型组、滋肾活血高剂量组、滋肾活血中剂量组、滋肾活血低剂量组、西药组。除假手术组外,其余各组均采用改良2-VO法建立VD大鼠模型。每组大鼠按9 mL/(kg·d)剂量灌胃相应药物。模型组、假手术组给予蒸馏水,滋肾活血低剂量组、滋肾活血中剂量组、滋肾活血高剂量组予以滋肾活血方溶液[9.8、17.8、35.6 g/(kg·d)]灌胃,西药组以多奈哌齐溶液[150 mg/(kg·d)]灌胃。连续喂药2周后,采用水迷宫实验评估大鼠学习记忆功能;取海马组织,采用免疫组化法检测实验大鼠海马组织中的Mfn1、Mfn2、Drp1、Fis1蛋白表达量。结果与假手术组比较,模型组大鼠逃避潜伏期(escape latency,EL)延长(P<0.05),Mfn1、Mfn2、Drp1蛋白的表达量降低(P<0.05)。与模型组对比,滋肾活血中、高剂量组大鼠EL缩短(P<0.05),Mfn1、Mfn2、Drp1蛋白表达量升高(P<0.05)。与滋肾活血低剂量组比较,滋肾活血中、高剂量组大鼠EL缩短(P<0.05),Mfn1、Drp1蛋白表达量升高(P<0.05)。结论滋肾活血方可能通过调节细胞内线粒体分裂与融合,上调Mfn1、Mfn2、Drp1蛋白的表达,从而改善认知功能。

Objective To observe the effects of Zishen Huoxue Formula on the expressions of mitofusin 1(Mfn1),mitofusin 2(Mfn2),dynamin-related protein 1(Drp1)and fission mitochondrial 1(Fis1)in hippocampus of rats with vascular dementia(VD).Methods A total of 60 male SD rats were randomly divided into sham operation group,model group,Zishen Huoxue high-dose group,Zishen Huoxue medium-dose group,Zishen Huoxue low-dose group and western medicine group.Except the sham operation group,the VD rat models were established by modified 2-VO method in the other groups.Rats in each group were given the corresponding drug by gavage at a dose of 9 mL/(kg·d).The model group and the sham operation group were given distilled water,the Zishen Huoxue low-dose group,Zishen Huoxue medium-dose group and the Zishen Huoxue high dose group were given solution of Zishen Huoxue Formula[9.8,17.8,35.6 g/(kg·d)],and the western medicine group was given donepezil solution[150 mg/(kg·d)].After 2 weeks of continuous administration,the learning and memory function of rats was evaluated by the Morris water maze,and then the protein expressions of Mfn1,Mfn2,Drp1 and Fis1 in the hippocampi of the experimental rats were detected by immunohistochemistry.Results Compared with the sham operation group,the escape latency(EL)of the model group was prolonged(P<0.05),and the expressions of Mfn1,Mfn2,Drp1 proteins were lower(P<0.05).Compared with the model group,the escape latency of Zishen Huoxue medium-dose group and Zishen Huoxue high-dose group was reduced,and the expressions of Mfn1,Mfn2,Drp1 proteins were higher(P<0.05).Compared with the Zishen Huoxue low-dose group,the escape latency of the medium-dose and high-dose group was reduced(P<0.05),and the expressions of Mfn2 and Drp1 were higher(P<0.05).Conclusion Zishen Huoxue Formula may improve cognitive function by regulating the mitochondrial fission and fusion in cells,and up-regulating the expressions of Mfn1,Mfn2 and Drp1 proteins.