摘要
目的探讨Rho相关卷曲螺旋形成蛋白激酶(Rho-associated coiled-coil forming prote in kinase,ROCK)-Ⅰ选择性阻断剂法舒地尔对大鼠肝纤维化组织ROCK-Ⅰ、α-SMA和p-MBSThr-697表达及细胞骨架变化的影响。方法应用Westernblot方法测定肝组织ROCK-Ⅰ、α-SMA和p-MBSThr-697表达,RT-PCR方法测定肝组织ROCK-Ⅰ和α-SMAmRNA表达,特异性免疫荧光方法检测细胞骨架变化。结果①Western blot结果显示,胆总管结扎(bileduct ligation,BDL)┼法舒地尔腹腔注射组造模第4周ROCK-Ⅰ蛋白、p-MBSThr-697蛋白和α-SMA蛋白与BDL组相比分别降低了38.68%、30.69%和20.50%。②RT-PCR结果显示,BDL┼法舒地尔腹腔注射组造模第4周ROCK-ⅠmRNA和α-SMA mRNA与BDL组相比分别降低了32.39%和34.46%。③FITC标记的鬼笔毒环肽荧光染色结果显示,BDL┼法舒地尔腹腔注射组与BDL组相比,可见大鼠肝组织细胞间网状结构稍规则,周边肌动蛋白丝带连续,荧光信号减弱。结论 ROCK-Ⅰ选择性阻断剂法舒地尔可抑制BDL大鼠模型肝组织ROCK-Ⅰ、p-MBSThr-697和α-SMA转录及翻译水平表达,可部分抑制BDL大鼠肝组织F-actin重构。
Objective To investigate the effects of Rho-associated coiled-coil forming protein kinase （ROCK）-I inhibitor, fasudil on the expression of ROCK-Ⅰ, α-SMA and p-MBS Thr-697 in liver fibrosis tissue and the changes of cytoskeleton. Methods The expressions of ROCK-Ⅰ, α-SMA and p-MBS Thr-697 protein in the liver were determined by Western blot. ROCK-Ⅰ, α-SMA mRNA were examined by reverse transcription-polymerase chain reaction （RT-PCR）. The changes of actin cytoskeleton were studied by immunofluorescence assay. Results The expressions of ROCK-Ⅰ, α-SMA and p-MBS Thr-697 protein at week 4 in BDL＋fasudil group decreased by 38.68%, 30.69% and 20.50%, respectively. RT-PCR showed that the expressions of ROCK-Ⅰ, α-SMA mRNA at week 4 in BDL^fasudil group reduced by 32.39% and 34.46%, respectively. Compared with BDL group, peripheral filament band of liver tissue became regular, actin became continuous and fluorescent signal was weaken in BDL＋fasudil group. Conclusions ROCK-Ⅰ inhibitor, fasudil may inhibit the expressions of transcription and translation levels of ROCK-Ⅰ, p-MBS Thr-697 and α-SMA in liver tissue of BDL rat model, and can partly inhibit recombination of F-aetin.
出处
《传染病信息》
2011年第3期140-142,170,共4页
Infectious Disease Information
基金
河北省自然科学基金(C2007000843)
