【摘要】：目的:通過研究辛二酰苯胺異羥肟酸(SAHA)對大鼠原代肝星狀細胞(HSCs)凋亡及相關(guān)蛋白表達的影響,探討SAHA誘導HSCs凋亡的作用機制。方法:采用Opti Prep梯度離心法分離大鼠原代HSCs;通過實時細胞分析技術(shù)檢測SAHA對HSCs增殖的影響;倒置顯微鏡觀察不同濃度SAHA處理HSCs后的形態(tài)變化;熒光顯微鏡及流式細胞術(shù)Annexin V-FITC/PI法檢測細胞凋亡率;Western blotting法檢測α-平滑肌肌動蛋白(α-SMA)、Ⅰ型膠原、金屬蛋白酶組織抑制物1(TIMP1)、葡萄糖調(diào)節(jié)蛋白78(GRP78)和組蛋白去乙�；�6(HDAC6)的蛋白表達;免疫共沉淀法檢測GRP78蛋白與HDAC6蛋白是否形成復合物。結(jié)果:成功分離的HSCs連續(xù)培養(yǎng)14 d,HSCs逐漸由靜止狀態(tài)變?yōu)榧せ顮顟B(tài)。SAHA可顯著抑制HSCs增殖,且呈劑量時間依賴性(P0.05);SAHA對HSCs的促凋亡作用具有時間依賴性(P0.05)。SAHA處理HSCs后,α-SMA、Ⅰ型膠原、HDAC6和TIMP1的蛋白表達水平明顯降低(P0.05),GRP78的蛋白表達水平明顯升高(P0.05)。與激活型的HSCs相比,SAHA處理后的HSCs中免疫共沉淀下的蛋白復合物中GRP78與總的乙�；嚢彼岬鞍罪@著增多,而HDAC6蛋白顯著降低,同時證明GRP78與HDAC6形成復合物。結(jié)論:SAHA抗肝纖維化的機制可能是,SAHA下調(diào)HDAC6蛋白表達水平,上調(diào)乙�；疓RP78蛋白表達水平,誘導HSCs內(nèi)質(zhì)網(wǎng)應激,促進肝星狀細胞凋亡,從而起到抗肝纖維化的作用。
[Abstract]:Aim: to investigate the effect of (SAHA) on (HSCs) apoptosis and related protein expression in rat primary hepatic stellate cells, and to explore the mechanism of HSCs apoptosis induced by SAHA. Methods: primary rat HSCs were isolated by Opti Prep gradient centrifugation, the effects of SAHA on HSCs proliferation were detected by real-time cell analysis, morphological changes of HSCs treated with different concentrations of SAHA were observed by inverted microscope. The expression of 偽 -smooth muscle actin (偽 -SMA), type 鈪，

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