摘要
目的探讨HMGB1在肺泡上皮细胞间质转分化中的作用。方法设计并合成针对HMGB1的小干扰RNA(siRNA),体外培养Ⅱ型肺泡上皮细胞系细胞-A549细胞,将细胞分为4组:1)对照组,2)TGF-β1刺激组,3)RNAi组(TGF-β1+HMGB1 siRNA),4)RNAi阴性对照组(TGF-β1+siRNA阴性对照);倒置相差显微镜观察细胞形态学;RT-PCR法检测HMGB1和α平滑肌肌动蛋白(α-SMA)的mRNA表达;Western blot法检测α-SMA和HMGB1蛋白表达。结果 TGF-β1刺激组细胞HMGB1和α-SMA的mRNA和蛋白表达均较对照组显著增加(P<0.01);RNAi组HMGB1、α-SMA mRNA和蛋白表达均较正常对照组显著下降(P<0.01)。结论 HMGB1可能参与了TGFβ1诱导的肺泡上皮细胞转分化。
Objective To investigate the role of HMGB1 in epithelial-mesenchymal transition. Methods Specific siRNA of HMGB1 was designed and synthesized. Cultured typeⅡ alveolar epithelial cell line-A549 cells were divided into 4 groups： 1） control group,2） model group induced by TGF-β1,3） HMGB1 RNAi group,4） RNAi negative control group. Cellular morphology changes were observed by phase-contrast microscope. HMGB1 and α-SMA expression in A549 cells was detected by RT-PCR and Western blotting respectively. Results mRNA and protein expression of HMGB1 and α-SMA in TGF-β1 group increased significantly than that in control group（ P〈 0. 01）. HMGB1,α-SMA mRNA and protein expression in siRNA-treated cells decreased significantly as compared with that in TGF-β1 group（ P〈 0. 01）. Conclusions HMGB1 may be involved in the TGF-β 1 induced EMT.
出处
《基础医学与临床》
CSCD
2015年第2期183-186,共4页
Basic and Clinical Medicine
基金
广东省医学科研基金(B2013068)
广东食品药品职业学院自然科学研究项目(2012YZ006)
