中华实验外科杂志
中華實驗外科雜誌
중화실험외과잡지
CHINESE JOURNAL OF EXPERIMENTAL SURGERY
2013年
5期
1002-1004
,共3页
周燕%魏捷%梁远红%陈静%唐其柱
週燕%魏捷%樑遠紅%陳靜%唐其柱
주연%위첩%량원홍%진정%당기주
p38丝裂原活化蛋白激酶%慢病毒%结缔组织生长因子%RNA 干扰
p38絲裂原活化蛋白激酶%慢病毒%結締組織生長因子%RNA 榦擾
p38사렬원활화단백격매%만병독%결체조직생장인자%RNA 간우
p38 mitogen-activated protein kinase%Lentiviral vectors%Connective tissue growth factor%RNA interference
目的 观察慢病毒介导的p38丝裂原活化蛋白激酶(p38MAPK)的短发夹环RNA(shRNA)对心肌成纤维细胞结缔组织生长因子(CTGF)的影响并探讨其机制.方法 构建慢病毒p38MAPK shRNA (PGLV-shRNA)并测序鉴定,观察其对转化生长因子-β(TGF-β)诱导心肌成纤维细胞的影响,检测p38MAPK、CTGF mRNA和CTGF、α-平滑肌肌动蛋白(α-SMA)蛋白的表达.结果 TGF-β3 4 nmol/L刺激24 h使心肌成纤维细胞p38MAPK mRNA、CTGF mRNA、CTGF蛋白和α-SMA蛋白明显升高(P＜0.01).PGLV-shRNA明显减少TGF-β诱导的p38MAPK mRNA、CTGF mRNA、CTGF蛋白和α-SMA蛋白表达(0.252±0.041比0.652±0.089,P＜0.01；0.418 ±0.071比0.838±0.099,P ＜0.01;0.418 ±0.076比0.991 ±0.117,P＜ 0.01;0.465 ±0.069比0.875 ±0.100,P＜0.05).结论 TGF-β经p38MAPK信号通路诱导心肌成纤维细胞CTGF表达,PGLV-shRNA能有效抑制TGF-β诱导的心肌细胞纤维化.
目的 觀察慢病毒介導的p38絲裂原活化蛋白激酶(p38MAPK)的短髮夾環RNA(shRNA)對心肌成纖維細胞結締組織生長因子(CTGF)的影響併探討其機製.方法 構建慢病毒p38MAPK shRNA (PGLV-shRNA)併測序鑒定,觀察其對轉化生長因子-β(TGF-β)誘導心肌成纖維細胞的影響,檢測p38MAPK、CTGF mRNA和CTGF、α-平滑肌肌動蛋白(α-SMA)蛋白的錶達.結果 TGF-β3 4 nmol/L刺激24 h使心肌成纖維細胞p38MAPK mRNA、CTGF mRNA、CTGF蛋白和α-SMA蛋白明顯升高(P＜0.01).PGLV-shRNA明顯減少TGF-β誘導的p38MAPK mRNA、CTGF mRNA、CTGF蛋白和α-SMA蛋白錶達(0.252±0.041比0.652±0.089,P＜0.01；0.418 ±0.071比0.838±0.099,P ＜0.01;0.418 ±0.076比0.991 ±0.117,P＜ 0.01;0.465 ±0.069比0.875 ±0.100,P＜0.05).結論 TGF-β經p38MAPK信號通路誘導心肌成纖維細胞CTGF錶達,PGLV-shRNA能有效抑製TGF-β誘導的心肌細胞纖維化.
목적 관찰만병독개도적p38사렬원활화단백격매(p38MAPK)적단발협배RNA(shRNA)대심기성섬유세포결체조직생장인자(CTGF)적영향병탐토기궤제.방법 구건만병독p38MAPK shRNA (PGLV-shRNA)병측서감정,관찰기대전화생장인자-β(TGF-β)유도심기성섬유세포적영향,검측p38MAPK、CTGF mRNA화CTGF、α-평활기기동단백(α-SMA)단백적표체.결과 TGF-β3 4 nmol/L자격24 h사심기성섬유세포p38MAPK mRNA、CTGF mRNA、CTGF단백화α-SMA단백명현승고(P＜0.01).PGLV-shRNA명현감소TGF-β유도적p38MAPK mRNA、CTGF mRNA、CTGF단백화α-SMA단백표체(0.252±0.041비0.652±0.089,P＜0.01；0.418 ±0.071비0.838±0.099,P ＜0.01;0.418 ±0.076비0.991 ±0.117,P＜ 0.01;0.465 ±0.069비0.875 ±0.100,P＜0.05).결론 TGF-β경p38MAPK신호통로유도심기성섬유세포CTGF표체,PGLV-shRNA능유효억제TGF-β유도적심기세포섬유화.
Objective To explore the effects of p38 mitogen-activated protein kinase (MAPK)short-hair RNA (shRNA) delivered by lentiviral vectors (PGLV) on connective tissue growth factor (CTGF) expression in myocardial fibroblasts.Methods The PGLN-shRNA was transfected into myocardial fibroblasts to to explore the role of p38MAPK pathway activation in transforming growth factor (TGF)-β-mediated myocardial fibrosis.The mRNA expression levels of p38MAPK and CTGF were detected by using reverse transcriptase-polymerase chain reaction (RT-PCR),and the protein expression levels of CTGF and α-smooth muscle actin (α-SMA) by using Western blotting.Results TGF-β stimulation increased p38MAPK and CTGF mRNA,and CTGF and α-SMA protein expression in myocardial fibroblasts (P ＜0.01).As compared with those in TGF-β group,PGLN-shRNA transfection decreased p38MAPK and CTGF mRNA,and CTGF and α-SMA protein expression in myocardial fibroblasts [(0.252 ± 0.041 vs.0.652±0.089,P＜0.01; 0.418 ±0.071 vs.0.838 ±0.099,P＜0.01; 0.418 ±0.076 vs.0.991 ±0.117,P＜0.01;0.465±0.069 vs.0.875 ± 0.100,P ＜ 0.05)].Conclusion TGF-β directly upregulates CTGF expression through p38MAPK pathway and gene silence of p38MAPK may protect TGF-β-mediated fibrosis in cardiac myoeytes.
