雷帕霉素减轻氧糖剥夺对SH-SY5Y细胞的损伤

发布时间：2018-03-29 20:24

  本文选题：雷帕霉素　切入点：SH-SYY细胞　出处：《中国病理生理杂志》2017年01期

【摘要】：目的:观察雷帕霉素(Rapa)对氧糖剥夺(OGD)的人神经母细胞瘤SH-SY5Y细胞的影响,并探讨自噬在其中的作用。方法:SH-SY5Y细胞随机分为4组:正常对照组(常规培养,不进行OGD处理)、Rapa组、OGD组(无糖培养基、1%O_2的三气培养箱内孵育细胞12 h)和Rapa+OGD组。进行形态学观察;MTT法检测细胞活力;乳酸脱氢酶(LDH)漏出率判断细胞损伤的程度;caspase-3活性检测试剂盒检测酶活性;原位末端标记(TUNEL)法检测凋亡水平;Western blot法检测凋亡相关蛋白Bax和Bcl-2、自噬标志蛋白LC3B-Ⅱ及自噬调控蛋白beclin-1的表达。结果:与OGD组相比,Rapa+OGD组的细胞存活率明显升高(P0.05),LDH漏出率及caspase-3酶活性明显降低(P0.05)。TUNEL染色观察结果显示,与OGD组相比,Rapa+OGD组的细胞凋亡明显减少(P0.05);Western blot实验结果显示Rapa+OGD组的Bcl-2、beclin-1及LC3B-Ⅱ蛋白的表达水平显著高于OGD组(P0.05),而Bax蛋白水平明显低于OGD组(P0.05)。结论:Rapa对OGD损伤的SH-SY5Y细胞具有保护作用,其机制可能与上调beclin-1蛋白、激活自噬有关。
[Abstract]:Objective: to observe the effect of rapamycin Rapa on SH-SY5Y cells of human neuroblastoma with oxygen glucose deprivation and to explore the role of autophagy. Methods: SH-SY5Y cells were randomly divided into 4 groups: normal control group (conventional culture). The cells were incubated with OGD for 12 hours in the three-air incubator without OGD in Rapa group and Rapa OGD group. The viability of the cells was detected by morphological observation. The leakage rate of lactate dehydrogenase (LDH) was used to determine the degree of cell injury and the activity of caspase-3 was detected by the kit. The expression of apoptosis-related protein Bax and Bcl-2, autophagy marker LC3B- 鈪，

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