中华医学杂志
中華醫學雜誌
중화의학잡지
National Medical Journal of China
2013年
19期
1490-1493
,共4页
李凤贺%赵渝%王学虎%时德%傅麒宁%刘洪
李鳳賀%趙渝%王學虎%時德%傅麒寧%劉洪
리봉하%조투%왕학호%시덕%부기저%류홍
血小板源性生长因子%肌细胞,平滑肌%增殖%sonic hedgehog
血小闆源性生長因子%肌細胞,平滑肌%增殖%sonic hedgehog
혈소판원성생장인자%기세포,평활기%증식%sonic hedgehog
Platelet-derived growth factor%Myocytes,smooth muscle%Proliferation%SHH
目的 探讨sonic hedgehog (SHH)信号通路在血小板源性生长因子(PDGF)诱导的血管平滑肌细胞(VSMC)增殖中的作用.方法 体外培养人大隐静脉VSMC,施加PDGF刺激后应用激光共聚焦显微成像及蛋白印迹(Western blot)检测SHH信号通路分子mRNA的表达变化；分别应用慢病毒介导的RNAi技术及SHH信号通路特异抑制剂cyclopamine阻断SHH通路后,用BrdU掺入法联合Ki-67染色评价细胞增殖情况.结果 激光共聚焦显微成像发现与对照组相比,PDGF刺激能够显著诱导VSMC中SHH信号通路蛋白shh,Patched1及Gli2的表达；同时,蛋白印迹也显示PDGF刺激能够明显增加SHH信号通路分子的表达；Gli2-siRNA慢病毒载体能够抑制Gli2的表达,Gli2-siRNA组BrdU掺入量为对照组的(49±0.6)％；与对照组相比,Gli2-siRNA组Ki-67阳性细胞数也明显减少；蛋白印迹显示SHH信号通路特异抑制剂cyclopamine能够抑制SHH信号通路下游转录因子Gli2的表达；cyclopamine组BrdU掺入量为对照组的(52±0.3)％,与对照组相比,cyclopamine组Ki-67阳性细胞数明显减少.结论 SHH信号通路在PDGF诱导的VSMC增殖中有重要作用.
目的 探討sonic hedgehog (SHH)信號通路在血小闆源性生長因子(PDGF)誘導的血管平滑肌細胞(VSMC)增殖中的作用.方法 體外培養人大隱靜脈VSMC,施加PDGF刺激後應用激光共聚焦顯微成像及蛋白印跡(Western blot)檢測SHH信號通路分子mRNA的錶達變化；分彆應用慢病毒介導的RNAi技術及SHH信號通路特異抑製劑cyclopamine阻斷SHH通路後,用BrdU摻入法聯閤Ki-67染色評價細胞增殖情況.結果 激光共聚焦顯微成像髮現與對照組相比,PDGF刺激能夠顯著誘導VSMC中SHH信號通路蛋白shh,Patched1及Gli2的錶達；同時,蛋白印跡也顯示PDGF刺激能夠明顯增加SHH信號通路分子的錶達；Gli2-siRNA慢病毒載體能夠抑製Gli2的錶達,Gli2-siRNA組BrdU摻入量為對照組的(49±0.6)％；與對照組相比,Gli2-siRNA組Ki-67暘性細胞數也明顯減少；蛋白印跡顯示SHH信號通路特異抑製劑cyclopamine能夠抑製SHH信號通路下遊轉錄因子Gli2的錶達；cyclopamine組BrdU摻入量為對照組的(52±0.3)％,與對照組相比,cyclopamine組Ki-67暘性細胞數明顯減少.結論 SHH信號通路在PDGF誘導的VSMC增殖中有重要作用.
목적 탐토sonic hedgehog (SHH)신호통로재혈소판원성생장인자(PDGF)유도적혈관평활기세포(VSMC)증식중적작용.방법 체외배양인대은정맥VSMC,시가PDGF자격후응용격광공취초현미성상급단백인적(Western blot)검측SHH신호통로분자mRNA적표체변화；분별응용만병독개도적RNAi기술급SHH신호통로특이억제제cyclopamine조단SHH통로후,용BrdU참입법연합Ki-67염색평개세포증식정황.결과 격광공취초현미성상발현여대조조상비,PDGF자격능구현저유도VSMC중SHH신호통로단백shh,Patched1급Gli2적표체；동시,단백인적야현시PDGF자격능구명현증가SHH신호통로분자적표체；Gli2-siRNA만병독재체능구억제Gli2적표체,Gli2-siRNA조BrdU참입량위대조조적(49±0.6)％；여대조조상비,Gli2-siRNA조Ki-67양성세포수야명현감소；단백인적현시SHH신호통로특이억제제cyclopamine능구억제SHH신호통로하유전록인자Gli2적표체；cyclopamine조BrdU참입량위대조조적(52±0.3)％,여대조조상비,cyclopamine조Ki-67양성세포수명현감소.결론 SHH신호통로재PDGF유도적VSMC증식중유중요작용.
Objective To discuss the role of sonic hedgehog (SHH) pathway in PDGF-induced vascular smooth muscle cell proliferation.Method Human vein VSMC were cultued in vitro.Laser confocal microscopy and Western blot were used to detect the expression of SHH pathway related proteins.The cell proliferation was evaluated by Ki-67 staining and BrdU incorporation after treatment by siRNA of Gli2 or shh pathway inhibitor cyclopamine.Results The results of laser confocal microscopy and Western blot showed that SHH pathway protein which including shh,Patchedl and G1i2 were activated in the PDGF-induced VSMC proliferation.BrdU incorporation assay and Ki-67 stianing showed that the cell proliferation which induced by PDGF was inhibited by Gli2-siRNA and cyclopamine which can both block SHH pathway.Conclusion SHH pathway play an important role in PDGF-induced VSMC proliferation.
