中国医学科学院学报
中國醫學科學院學報
중국의학과학원학보
ACTA ACADEMIAE MEDICINAE SINICAE
2010年
1期
32-38,后插1
,共8页
苗立夫%黄超联%陈连凤%朱文玲%杨菁%王以光%张华%刘佩毛%佘铭鹏%宋存先
苗立伕%黃超聯%陳連鳳%硃文玲%楊菁%王以光%張華%劉珮毛%佘銘鵬%宋存先
묘립부%황초련%진련봉%주문령%양정%왕이광%장화%류패모%사명붕%송존선
雷帕霉素%聚乳酸-聚乙醇酸共聚物%纳米粒子%人脐动脉平滑肌细胞%细胞周期%细胞增殖%p27蛋白
雷帕黴素%聚乳痠-聚乙醇痠共聚物%納米粒子%人臍動脈平滑肌細胞%細胞週期%細胞增殖%p27蛋白
뢰파매소%취유산-취을순산공취물%납미입자%인제동맥평활기세포%세포주기%세포증식%p27단백
rapamycin%poly(lactic-co-glycolic)acid%nanoparticles%human umbilical arterial vascular smooth muscle cell%cell cycle%proliferation%p27 protein
目的 评估本实验室自制包载雷帕霉素(RPM)的聚乳酸-聚乙醇酸共聚物(PLGA)纳米粒子(NPs)对离体培养的人脐动脉平滑肌细胞(HUASMC)细胞周期时相、p27蛋白表达和细胞增殖的影响.方法 按不同浓度RPM-PLGA NPs设定药物作用组,并设立RPM组、PLGA组和M231培养基及平滑肌细胞生长添加剂(M231-SMGs)组作为对照.采用细胞免疫组织化学染色比较不同处理组HUASMC p27蛋白表达阳性率和表达水平的差异,流式细胞技术评价各处理组对HUASMC细胞周期时相的影响,噻唑蓝(MTT)比色法观察不同浓度RPM和RPM-PLGA NPs对HUASMC存活率的影响.结果 10μg/L及以上浓度的RPM和50 μg/L及以上浓度的RPM-PLGA NPs可明显抑制HUA-SMC生长,并呈浓度依赖性.细胞计数绘制生长-时间曲线显示,100μg/L RPM和500μg/L RPM-PLGA NPs作用于HUASMC 24h后细胞计数值明显低于M231-SMGs对照组;单纯PLGA NPs对细胞生长无明显影响;与PLGA组和M231-SMGs培养基对照组相比,RPM组和RPM-PLGA NPs组G_0/G_1期细胞比例明显增多,S期及G_2/M期细胞比例明显减少(P均<0.01),但两组间细胞周期时相比例差异无统计学意义(P>0.05).细胞免疫组织化学染色结果显示,100μg/LRPM和500μg/L RPM-PLGA NPs组的HUASMC p27蛋白表达阳性率和表达水平与对照组相比差异无统计学意义(P>0.05),但均较PLGA组和M231-SMGs培养基组明显增加(P<0.01).结论 RPM-PLGA NPs抑制体外培养的HUASMC生长效果与RPM相似,并可显著抑制体外培养HUASMC p27蛋白表达,阻抑其细胞周期进程于G1/S期而抑制其细胞增殖.
目的 評估本實驗室自製包載雷帕黴素(RPM)的聚乳痠-聚乙醇痠共聚物(PLGA)納米粒子(NPs)對離體培養的人臍動脈平滑肌細胞(HUASMC)細胞週期時相、p27蛋白錶達和細胞增殖的影響.方法 按不同濃度RPM-PLGA NPs設定藥物作用組,併設立RPM組、PLGA組和M231培養基及平滑肌細胞生長添加劑(M231-SMGs)組作為對照.採用細胞免疫組織化學染色比較不同處理組HUASMC p27蛋白錶達暘性率和錶達水平的差異,流式細胞技術評價各處理組對HUASMC細胞週期時相的影響,噻唑藍(MTT)比色法觀察不同濃度RPM和RPM-PLGA NPs對HUASMC存活率的影響.結果 10μg/L及以上濃度的RPM和50 μg/L及以上濃度的RPM-PLGA NPs可明顯抑製HUA-SMC生長,併呈濃度依賴性.細胞計數繪製生長-時間麯線顯示,100μg/L RPM和500μg/L RPM-PLGA NPs作用于HUASMC 24h後細胞計數值明顯低于M231-SMGs對照組;單純PLGA NPs對細胞生長無明顯影響;與PLGA組和M231-SMGs培養基對照組相比,RPM組和RPM-PLGA NPs組G_0/G_1期細胞比例明顯增多,S期及G_2/M期細胞比例明顯減少(P均<0.01),但兩組間細胞週期時相比例差異無統計學意義(P>0.05).細胞免疫組織化學染色結果顯示,100μg/LRPM和500μg/L RPM-PLGA NPs組的HUASMC p27蛋白錶達暘性率和錶達水平與對照組相比差異無統計學意義(P>0.05),但均較PLGA組和M231-SMGs培養基組明顯增加(P<0.01).結論 RPM-PLGA NPs抑製體外培養的HUASMC生長效果與RPM相似,併可顯著抑製體外培養HUASMC p27蛋白錶達,阻抑其細胞週期進程于G1/S期而抑製其細胞增殖.
목적 평고본실험실자제포재뢰파매소(RPM)적취유산-취을순산공취물(PLGA)납미입자(NPs)대리체배양적인제동맥평활기세포(HUASMC)세포주기시상、p27단백표체화세포증식적영향.방법 안불동농도RPM-PLGA NPs설정약물작용조,병설립RPM조、PLGA조화M231배양기급평활기세포생장첨가제(M231-SMGs)조작위대조.채용세포면역조직화학염색비교불동처리조HUASMC p27단백표체양성솔화표체수평적차이,류식세포기술평개각처리조대HUASMC세포주기시상적영향,새서람(MTT)비색법관찰불동농도RPM화RPM-PLGA NPs대HUASMC존활솔적영향.결과 10μg/L급이상농도적RPM화50 μg/L급이상농도적RPM-PLGA NPs가명현억제HUA-SMC생장,병정농도의뢰성.세포계수회제생장-시간곡선현시,100μg/L RPM화500μg/L RPM-PLGA NPs작용우HUASMC 24h후세포계수치명현저우M231-SMGs대조조;단순PLGA NPs대세포생장무명현영향;여PLGA조화M231-SMGs배양기대조조상비,RPM조화RPM-PLGA NPs조G_0/G_1기세포비례명현증다,S기급G_2/M기세포비례명현감소(P균<0.01),단량조간세포주기시상비례차이무통계학의의(P>0.05).세포면역조직화학염색결과현시,100μg/LRPM화500μg/L RPM-PLGA NPs조적HUASMC p27단백표체양성솔화표체수평여대조조상비차이무통계학의의(P>0.05),단균교PLGA조화M231-SMGs배양기조명현증가(P<0.01).결론 RPM-PLGA NPs억제체외배양적HUASMC생장효과여RPM상사,병가현저억제체외배양HUASMC p27단백표체,조억기세포주기진정우G1/S기이억제기세포증식.
Objective To evaluate the effects of rapamycin(RPM)-loaded poly(lactic-co-glycolic)acid(PLGA)nanoparticles(NPs)on the proliferation,distribution of cell cycle,and expression of p27 protein in human umbilical arterial vascular smooth muscle cell(HUASMC)in vitro.Methods The primarily culture model of HUASMC was successfully established by explant-attached method in vitro.The cells were administrated with different doses of BPM,and RPM-PLGA NPs were observed as treat groups compared with PLGA NPs and M231-SMGs medium cultured group.The effect of RPM-PLGA NPs on proliferation of HUASMC was assessed using 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide(MTT)colorimetry method.The influences of RPM-PLGA NPs on the cell cycle and cellular growth kinetics of HUASMCs were tested by flow cytometry.The effect of RPM-PLGA NPs on the expression of p27 protein of HUASMCs was assessed through an immunohistochemical method.Results Compared with the control group,the proliferation of HUASMCs was inhibited by 50μg/L and higher concentration of RPM-PLGA NPs in a dose-dependent manner(P < 0.05).The numbers of cells entering cell cycle of S/G_2/M phases were significantly lower in BPMPLGA NPs and RPM treated groups.Histologically,the expression of p27 were up-regulated in 500 μg/L RPMPLGA NPs and 100μg/L RPM treated group(all P <0.01)when compared with the control group.Conclusions RPM-PLGA NPs has a similar effects as RPM in inhibiting the growth of in vitro cultured HUASMC.It can remarkably suppress the expression of in vitro cultured HUASMC p27 protein,arrest its cell cycle at G_1/S phase,and inhibit its proliferation.
