本文選題：肝纖維化 + UBC　； 參考：《實(shí)用醫(yī)學(xué)雜志》2017年13期

【摘要】：目的研究類泛素綴和酶9(UBC9)在肝星狀細(xì)胞的激活及肝纖維化中的作用機(jī)制。方法通過蛋白印記法(Western blot)檢測UBC9在不同濃度的轉(zhuǎn)化生長因子-β1(TGF-β1)刺激下的肝星狀細(xì)胞LX-2中的表達(dá)情況;構(gòu)建UBC9質(zhì)粒干擾片段(UBC9-shRNA),采用脂質(zhì)體介導(dǎo)轉(zhuǎn)染LX-2,同時(shí)設(shè)置無義序列組和空白對(duì)照組,實(shí)時(shí)熒光定量PCR檢測轉(zhuǎn)染后UBC9 mRNA的表達(dá);Western blot檢測UBC9在蛋白水平的表達(dá);Western blot檢測下調(diào)UBC9對(duì)α-SMA、Collagen Ⅰ及磷酸化的Smad3表達(dá)的影響。CCK8檢測轉(zhuǎn)染后細(xì)胞增殖能力。結(jié)果 UBC9在TGF-β1因子刺激下的肝星狀細(xì)胞LX-2表達(dá)增加,一定劑量下呈濃度依賴性。轉(zhuǎn)染干擾片段UBC9-shRNA后,激活的LX-2細(xì)胞增殖減弱,α-SMA和Collagen Ⅰ蛋白的表達(dá)減少,而且下調(diào)UBC9也能抑制磷酸化的Smad3的表達(dá)。結(jié)論下調(diào)UBC9可以通過抑制TGF-β1/Smad3信號(hào)通路進(jìn)而影響肝星狀細(xì)胞LX-2的激活,這些都提示UBC9在肝纖維化過程具有重要作用,為肝纖維化治療提供一些新的理論依據(jù)。
[Abstract]:Objective to study the role of UBC9 and UBC9 in the activation and fibrosis of hepatic stellate cells. Methods the expression of UBC9 in hepatic stellate cells stimulated by TGF- 尾 1 at different concentrations was detected by Western blot. UBC9 plasmid interference fragment UBC9-shRNA was constructed and transfected into LX-2 by liposome-mediated transfection. At the same time, nonsense sequence group and blank control group were set up. The expression of UBC9 mRNA after transfection was detected by real-time fluorescence quantitative PCR. Western blot was used to detect the expression of UBC9 at protein level. Western blot down-regulated the effect of UBC9 on the expression of 偽 -SMA-Collagen 鈪，

本文編號(hào)：1858941