發(fā)布時間：2018-03-29 20:24

  本文選題：雷帕霉素　切入點(diǎn)：SH-SYY細(xì)胞　出處：《中國病理生理雜志》2017年01期

【摘要】：目的:觀察雷帕霉素(Rapa)對氧糖剝奪(OGD)的人神經(jīng)母細(xì)胞瘤SH-SY5Y細(xì)胞的影響,并探討自噬在其中的作用。方法:SH-SY5Y細(xì)胞隨機(jī)分為4組:正常對照組(常規(guī)培養(yǎng),不進(jìn)行OGD處理)、Rapa組、OGD組(無糖培養(yǎng)基、1%O_2的三氣培養(yǎng)箱內(nèi)孵育細(xì)胞12 h)和Rapa+OGD組。進(jìn)行形態(tài)學(xué)觀察;MTT法檢測細(xì)胞活力;乳酸脫氫酶(LDH)漏出率判斷細(xì)胞損傷的程度;caspase-3活性檢測試劑盒檢測酶活性;原位末端標(biāo)記(TUNEL)法檢測凋亡水平;Western blot法檢測凋亡相關(guān)蛋白Bax和Bcl-2、自噬標(biāo)志蛋白LC3B-Ⅱ及自噬調(diào)控蛋白beclin-1的表達(dá)。結(jié)果:與OGD組相比,Rapa+OGD組的細(xì)胞存活率明顯升高(P0.05),LDH漏出率及caspase-3酶活性明顯降低(P0.05)。TUNEL染色觀察結(jié)果顯示,與OGD組相比,Rapa+OGD組的細(xì)胞凋亡明顯減少(P0.05);Western blot實驗結(jié)果顯示Rapa+OGD組的Bcl-2、beclin-1及LC3B-Ⅱ蛋白的表達(dá)水平顯著高于OGD組(P0.05),而Bax蛋白水平明顯低于OGD組(P0.05)。結(jié)論:Rapa對OGD損傷的SH-SY5Y細(xì)胞具有保護(hù)作用,其機(jī)制可能與上調(diào)beclin-1蛋白、激活自噬有關(guān)。
[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，

本文編號：1682705