犀角地黄汤对脑缺血大鼠的自噬相关蛋白Atg-5、Beclin-1表达的研究
Research of Xijiao Dihuang Decoction on expression of autophagy related protein Atg-5 and Beclin-1 in rats with cerebral ischemia
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摘要: 目的 探讨犀角地黄汤对内毒素诱导下持续性大脑中动脉栓塞模型(pMCAO)大鼠脑组织的自噬相关Atg-5、Beclin-1蛋白的mRNA表达,以及透射电镜下观察染色的LC-3抗体表达。方法 造模后进行分组,并采用不同剂量犀角地黄汤进行干预,分别提取RNA、PCR检测Rat-atg-5、Beclin-1基因表达,透射电镜下观察染色的LC-3抗体表达。结果 “瘀热”证pMCAO模型大鼠中Atg-5、Beclin-1 mRNA、模型组Atg-5和Beclin-1 mRNA明显上升,犀角地黄汤中剂量治疗组和阳性药组与模型组比较均显著降低其表达。LC-3抗体染色颗粒提示,空白组表达不明显,模型组造模成功,尤其在第6天,模型组明显增多;阳性药组明显抑制LC-3的表达,第3和第6天到达最佳;低剂量组抑制效果不明显;中剂量组抑制效果好于高剂量组,第6天抑制效果最佳。结论 犀角地黄汤通过显著的降低Atg-5、Beclin-1 mRNA的表达,以及明显抑制LC-3的表达,能够起到脑保护的作用。Abstract: OBJECTIVE To investigate the Xijiao Dihuang Decoction on mRNA expression of autophagy related Atg-5、Beclin-1 protein of the rats' brain tissues, which are endotoxin induced persistent middle cerebral artery occlusion (pMCAO) and to observe the anti-body expression of LC-3 through transmission electron microscope. METHODS Dived the rats into different groups after modeling, and then use different dose of Xijiao Dihuang Decoction for intervention. Extract RNA and use PCR to detect the gene expression of Rat-atg-5 and Beclin -1. The antibody expression of stained LC-3 was observed through transmission electron microscope. RESULTS Atg-5, Beclin-1 and mRNA significantly increased in "Stasis heat" syndrome rat model of "pMCAO" and the model group. The expression was significantly reduced Xijiao Dihuang Decoction middle dose group and positive drug group compared with the model group. LC-3 antibody staining granules suggested that the blank group did not have obvious expression. The model group rats, especially in the sixth day, the expression of model group was significantly increased and positive drug group significantly inhibited LC-3 expression. The third and sixth day reached the best point. Low dose group showed no inhibitory effect. The middle dose group had better effect than high dose group and reached the best point on the sixth day. CONCLUSION Xijiao Dihuang Decoction can protect the brain by significantly reducing the expression of Atg-5, Beclin-1 mRNA and inhibiting the expression of LC-3.
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Key words:
- Xijiao Dihuang Decoction /
- autophagy /
- Atg-5 /
- Beclin-1 /
- LC-3
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