补阳还五汤对SCII模型大鼠细胞自噬相关蛋白影响的研究

Research on the Effect of Buyang Huanwu Decoction on Autophagy Related Proteins in SCII Model Rats

目的:探讨补阳还五汤对大鼠脊髓缺血再灌注损伤(SCII)后细胞自噬相关蛋白Beclin 1、微管相关蛋白1轻链3(LC3)、P62(死骨片1)影响的机制研究。方法:48只SD大鼠随机分为假手术组、模型组、观察组和3-MA组,模型组腹腔内注射生理盐水,观察组在造模的前三天给予补阳还五汤汤药灌胃,3-MA(3-甲基腺嘌呤)组腹腔注射3-MA注射液,然后夹闭腹主动脉40min开放,造模后,在缺血3h、6h后进行行为学评分、取材,进行免疫组化检测Beclin 1、LC3、P62的表达变化、透视电镜检测。结果:在缺血3h后、6h后,假手术组与其他三组在Beclin1、LC3和P62等方面比较有统计学意义(P<0.05),模型组、观察组和3-MA组三种蛋白的表达均上调。行为学评分、透视电镜检测与免疫组化结果基本一致,可以相互印证。结论:补阳还五汤可能会对细胞自噬进行双向调节,保护神经细胞。

Objective:To investigate the mechanism of Buyang Huanwu Decoction on the effects of Beclin 1,LC3(microtubule-assaiated protein 1 light chain 3),P62(sequestosome 1)on autophagy related protein after SCII(Spinal cord ischemia-reperfbsion injury)in rats.Methods:Forty-eight SD rats were randomly divided into sham operation group,model group,observation group and 3-MA group.The model group was injected with normal saline intraperitoneally.The observation group was given Buyang Huanwu Decoction by gavage three days before modeling.In the 3-MA group,3-MA injection was injected intraperitoneally,and then the abdominal aorta was clamped for 40 minutes to open.After modeling,behavioral scores and materials were obtained after 3 hours and 6 hours of ischemia.The expression changes of Beclin 1,LC3 and p62 were detected by immunohistochemistry and fluoroscopy electron microscopy.Results:After 3 hours and 6 hours of ischemia,the expression of Beclin 1,LC3 and p62 in the sham operation group was significantly higher than that in the other three groups(P<0.05).The expression of three proteins in the model group,the observation group and the 3-MA group were all up-regulated.The results of behavioral score and electron microscopy were consistent with those of immunohistochemistry.Conclusion:Buyang Huanwu Ddecoction may have bidirectional regulation on autophagy to protect nerve cells.