中华医学杂志
中華醫學雜誌
중화의학잡지
National Medical Journal of China
2015年
38期
3147-3149
,共3页
闫迪%顾宪民%姜淑娟%王玉红
閆迪%顧憲民%薑淑娟%王玉紅
염적%고헌민%강숙연%왕옥홍
人抗原%肌动蛋白类%肌细胞,平滑肌%支气管%血小板源性生长因子
人抗原%肌動蛋白類%肌細胞,平滑肌%支氣管%血小闆源性生長因子
인항원%기동단백류%기세포,평활기%지기관%혈소판원성생장인자
Human-antigens%Actins%Myocytes,smooth muscle%Bronchi%Platelet-derived growth factor
目的 观察人支气管平滑肌细胞(HBSMC)内mRNA结合蛋白人抗原R(HuR)对α-平滑肌肌动蛋白(α-SMA)表达的影响.方法 体外培养HBSMC,根据血小板源性生长因子(PDGF)对其刺激时间分别记为0、6、12、24 h组.采用实时荧光定量PCR法检测各组HuR与α-SMA mRNA的表达、Western印迹法检测各组HuR与α-SMA蛋白的表达.干扰RNA法观察抑制HuR蛋白表达后PDGF刺激下对细胞α-SMA蛋白表达的影响.结果 PDGF时间依懒性增强HuR的表达,0、6、12、24 h组全细胞HuR蛋白及mRNA相对表达量分别为0.23 ±0.09、0.42 ±0.11、0.93 ±0.21、1.37±0.28及1.00±0.00、1.09±0.03、1.16 ±0.03、1.27±0.02(均P＜0.05);各组α-SMA蛋白及mRNA相对表达量也呈时间依赖性增强(1.03±0.08、1.20±0.09、1.39±0.11、1.58±0.10及1.00±0.00、1.17 ±0.02、1.23±0.02、1.45±0.03,均P＜0.05).HuR特异地干扰RNA转染细胞后,HuR表达下调,PDGF诱导的α-SMA表达相应减弱.结论 PDGF可增加HBSMC中HuR及α-SMA的表达,HuR参与PDGF诱导的α-SMA表达过程.
目的 觀察人支氣管平滑肌細胞(HBSMC)內mRNA結閤蛋白人抗原R(HuR)對α-平滑肌肌動蛋白(α-SMA)錶達的影響.方法 體外培養HBSMC,根據血小闆源性生長因子(PDGF)對其刺激時間分彆記為0、6、12、24 h組.採用實時熒光定量PCR法檢測各組HuR與α-SMA mRNA的錶達、Western印跡法檢測各組HuR與α-SMA蛋白的錶達.榦擾RNA法觀察抑製HuR蛋白錶達後PDGF刺激下對細胞α-SMA蛋白錶達的影響.結果 PDGF時間依懶性增彊HuR的錶達,0、6、12、24 h組全細胞HuR蛋白及mRNA相對錶達量分彆為0.23 ±0.09、0.42 ±0.11、0.93 ±0.21、1.37±0.28及1.00±0.00、1.09±0.03、1.16 ±0.03、1.27±0.02(均P＜0.05);各組α-SMA蛋白及mRNA相對錶達量也呈時間依賴性增彊(1.03±0.08、1.20±0.09、1.39±0.11、1.58±0.10及1.00±0.00、1.17 ±0.02、1.23±0.02、1.45±0.03,均P＜0.05).HuR特異地榦擾RNA轉染細胞後,HuR錶達下調,PDGF誘導的α-SMA錶達相應減弱.結論 PDGF可增加HBSMC中HuR及α-SMA的錶達,HuR參與PDGF誘導的α-SMA錶達過程.
목적 관찰인지기관평활기세포(HBSMC)내mRNA결합단백인항원R(HuR)대α-평활기기동단백(α-SMA)표체적영향.방법 체외배양HBSMC,근거혈소판원성생장인자(PDGF)대기자격시간분별기위0、6、12、24 h조.채용실시형광정량PCR법검측각조HuR여α-SMA mRNA적표체、Western인적법검측각조HuR여α-SMA단백적표체.간우RNA법관찰억제HuR단백표체후PDGF자격하대세포α-SMA단백표체적영향.결과 PDGF시간의라성증강HuR적표체,0、6、12、24 h조전세포HuR단백급mRNA상대표체량분별위0.23 ±0.09、0.42 ±0.11、0.93 ±0.21、1.37±0.28급1.00±0.00、1.09±0.03、1.16 ±0.03、1.27±0.02(균P＜0.05);각조α-SMA단백급mRNA상대표체량야정시간의뢰성증강(1.03±0.08、1.20±0.09、1.39±0.11、1.58±0.10급1.00±0.00、1.17 ±0.02、1.23±0.02、1.45±0.03,균P＜0.05).HuR특이지간우RNA전염세포후,HuR표체하조,PDGF유도적α-SMA표체상응감약.결론 PDGF가증가HBSMC중HuR급α-SMA적표체,HuR삼여PDGF유도적α-SMA표체과정.
Objective To investigate the role of mRNA binding protein Human-antigen R (HuR) in the over-expression of α-Smooth muscle actin (ot-SMA) stimulated by Platelet-derived Growth Factor (PDGF) in cultured human bronchia smooth muscle cells.Methods Human bronchia smooth muscle cells cultured in vitro were divided into 0,6,12 and 24 h groups according to the time of PDGF treatment.Total HuR protein and total α-SMA protein expression were detected by Western blot.Total HuR mRNA and total α-SMA mRNA level were determined by quantitative real time-polymerase chain reaction.RNA interference technology was used to down-regulate HuR protein level to study the protective effect of HuR in PDGFstimulated α-SMA protein expression.Results PDGF up-regulated the expression of HuR in a timedependent manner.The relative expression levels of whole-cell HuR protein and mRNA in 0,6,12,24 h groups were0.23 ±0.09,0.42 ±0.11,0.93 ±0.21,1.37 ±0.28;1.00 ±0.00,1.09 ±0.03,1.16± 0.03,1.27 ± 0.02 (all P ＜ 0.05).The relative expression levels of α-SMA protein and mRNA in 0,6,12,24 h group also showed an increase trend marked in a time-dependent manner (1.03 ± 0.08,1.20 ± 0.09,1.39 ±0.11,1.58 ±0.10;1.00±0.00,1.17 ±0.02,1.23 ±0.02,1.45 ±0.03;all P＜0.05).Using RNA interference technology to down-regulate HuR protein level,there was a decrease in α-SMA protein expression.Conclusion PDGF stimulation can increase the expression of HuR and α-SMA in the smooth muscle cells,and HuR protein is involved in the expression of c-SMA protein stimulated by PDGF.
