补阳还五汤对大鼠SCII自噬相关蛋白影响的实验研究

Experimental Study on the Effect of Buyang Huanwu Decoction on the Autophagy Related Proteins of SCII in Rats

目的探讨补阳还五汤对大鼠脊髓缺血再灌注损伤(spinal cord ischemia-reperfbsion injury,SCII)后自噬蛋白Beclin 1、LC3、P62影响的机制研究。方法将48只SD大鼠随机均分为假手术组、模型组、观察组和自噬抑制剂3-甲基腺嘌呤(3-methyladenine,3-MA)组,每组12只大鼠,并进行造模。模型制备后,在缺血3、6 h后进行行为学评分、取材,免疫组织化学检测Beclin 1、LC3、P62表达变化;透射电镜观察自噬小体情况;Western blot检测Beclin 1、LC3及P62共3种蛋白的表达情况。结果各组大鼠脊髓损伤后行为学评分比较差异具有统计学意义(P<0.05)。免疫组织化学显示在各组缺血3、6 h后脊髓组织中自噬相关蛋白Beclin 1、LC3和P62表达组间比较差异均有统计学意义(P<0.05)。Western blot检测结果显示Beclin 1、LC3、P62蛋白在各组中差异明显。结论补阳还五汤通过对自噬系统进行双向调节,保护神经细胞。

Objective To study the effect mechanism of Buyang Huanwu Decoction on the autophagy related proteins Beclin 1,LC3 and P62 after spinal cord ischemia-reperfbsion injury(SCII) in rats.Methods A total of 48 SD rats were randomly divided into sham operation group, model group, observation group and 3-methyladenine(3-MA) group,12 cases in each group. Preparation of model, the behavioral scores and samples were obtained after 3 and 6 hours of ischemia.The change expression of Beclin 1,LC3 and P62 were detected by immunohistochemistry.The autophagosomes were observed by transmission electron microscopy. The expressions of Beclin 1, LC3 and P62 were detected by Western blot.Results The behavioral scores were significantly different in each group(P<0.05). The results of immunohistochemistry showed that the autophagy related proteins Beclin 1,LC3 and P62 were significantly different in each group after 3 and 6 hours of ischemia(P<0.05). The results of Western blot showed that the expressions of Beclin 1, LC3 and P62 were significantly different in each group.Conclusion Buyang Huanwu Decoction can protect nerve cells by two-way regulation of autophagy system.