中华结核和呼吸杂志
中華結覈和呼吸雜誌
중화결핵화호흡잡지
Chinese Journal of Tuberculosis and Respiratory Diseases
2012年
7期
511-514
,共4页
王小双%范志宇%王涛%谢亮%余莉%高正祥%刘瀚旻
王小雙%範誌宇%王濤%謝亮%餘莉%高正祥%劉瀚旻
왕소쌍%범지우%왕도%사량%여리%고정상%류한민
紫杉醇%肌细胞,平滑肌%细胞表型
紫杉醇%肌細胞,平滑肌%細胞錶型
자삼순%기세포,평활기%세포표형
Paclitaxel%Myocytes,smooth muscle%Phenotype
目的 探讨不同浓度的紫杉醇对大鼠肺血管平滑肌细胞增殖的抑制作用及其对细胞骨架和表型转化的影响.方法 采用MTT、[3H]-胸腺嘧啶掺入法检测不同药物浓度紫杉醇对血小板源性生长因子BB( PDGF-BB)诱导的血管平滑肌细胞增殖的影响;采用蛋白质免疫印迹法(Westernblot)对平滑肌α肌动蛋白及平滑肌22α蛋白(SM22α)表达活性进行检测;利用激光共聚焦显微镜观察F-肌动蛋白的改变.结果 与PDGF组比较,紫杉醇药物干预组细胞增殖受到明显抑制,四甲基偶氮唑盐微量酶反应比色法(MTT)检测抑制率分别为40.0%,30.0%,18.0% (P<0.01),[3H]-胸腺嘧啶掺入法测得的细胞增殖抑制率分别为45.4%,35.4%,21.6% (P <0.01),平滑肌α肌动蛋白和SM22α表达均升高,细胞骨架F-肌动蛋白的荧光强度明显下降.结论 在PDGF-BB存在的情况下,紫杉醇可剂量依赖性抑制血管平滑肌细胞的增殖,通过作用于微丝细胞骨架促进血管平滑肌细胞由合成型向收缩型转化,进而影响血管平滑肌细胞的增殖.
目的 探討不同濃度的紫杉醇對大鼠肺血管平滑肌細胞增殖的抑製作用及其對細胞骨架和錶型轉化的影響.方法 採用MTT、[3H]-胸腺嘧啶摻入法檢測不同藥物濃度紫杉醇對血小闆源性生長因子BB( PDGF-BB)誘導的血管平滑肌細胞增殖的影響;採用蛋白質免疫印跡法(Westernblot)對平滑肌α肌動蛋白及平滑肌22α蛋白(SM22α)錶達活性進行檢測;利用激光共聚焦顯微鏡觀察F-肌動蛋白的改變.結果 與PDGF組比較,紫杉醇藥物榦預組細胞增殖受到明顯抑製,四甲基偶氮唑鹽微量酶反應比色法(MTT)檢測抑製率分彆為40.0%,30.0%,18.0% (P<0.01),[3H]-胸腺嘧啶摻入法測得的細胞增殖抑製率分彆為45.4%,35.4%,21.6% (P <0.01),平滑肌α肌動蛋白和SM22α錶達均升高,細胞骨架F-肌動蛋白的熒光彊度明顯下降.結論 在PDGF-BB存在的情況下,紫杉醇可劑量依賴性抑製血管平滑肌細胞的增殖,通過作用于微絲細胞骨架促進血管平滑肌細胞由閤成型嚮收縮型轉化,進而影響血管平滑肌細胞的增殖.
목적 탐토불동농도적자삼순대대서폐혈관평활기세포증식적억제작용급기대세포골가화표형전화적영향.방법 채용MTT、[3H]-흉선밀정참입법검측불동약물농도자삼순대혈소판원성생장인자BB( PDGF-BB)유도적혈관평활기세포증식적영향;채용단백질면역인적법(Westernblot)대평활기α기동단백급평활기22α단백(SM22α)표체활성진행검측;이용격광공취초현미경관찰F-기동단백적개변.결과 여PDGF조비교,자삼순약물간예조세포증식수도명현억제,사갑기우담서염미량매반응비색법(MTT)검측억제솔분별위40.0%,30.0%,18.0% (P<0.01),[3H]-흉선밀정참입법측득적세포증식억제솔분별위45.4%,35.4%,21.6% (P <0.01),평활기α기동단백화SM22α표체균승고,세포골가F-기동단백적형광강도명현하강.결론 재PDGF-BB존재적정황하,자삼순가제량의뢰성억제혈관평활기세포적증식,통과작용우미사세포골가촉진혈관평활기세포유합성형향수축형전화,진이영향혈관평활기세포적증식.
Objective To investigate the effects of paclitaxel on the phenotypic modulation induced by platelet-derived growth factor ( PDGF-BB ) in rat pulmonary vascular smooth muscle cells ( PVSMC ).Methods The proliferation of PVSMC isolated from SD rats cultured in vitro was induced by PDGF-BB and then intervened by different concentration of paclitaxel.MTT and [ 3H ]-thymidine incorporation were used to detect the changes of cell proliferation.The expression level of alpha-smooth muscle-actin (SM-α-actin) and smooth muscle protein 22alpha (SM22t) were tested by Western blot.Confocal laser scanning microscopy was applied to observe the change of fluorescence intensity.Results Treatment with PDGF-BB for 24 hours results in a significant increase in [ 3H ]-thymidine incorporation and marked change in phenotype and cytoskeleton,Paclitaxel inhibited the proliferation of PVSMC induced by PDGF-BB,the inhibition rate was 45.4%,35.4%,21.6% ( P < 0.01 ) tested by [ 3 H ] -thymidine incorporation and 40.0%,30.0%,18.0%( P < 0.01 ) tested by MTT.Meanwhile,the paclitaxel promoted the expression level of SM-α-actin and SM22α.Fluorescence intensity of F-actin decreased significanthy.Conclusion Paclitaxel may play an important role in vascular remodeling by changing the phenotypes and cytoskeleton of VSMC stimulated by PDGF-BB.