PTEN抑制增生性瘢痕成纤维细胞转分化作用及其机制  

PTEN inhibits trans-differentiation of hypertrophic scar fibroblasts in vitro

作　　者：郭亮[1,2] 毕胜[1] 王珍祥[1] 陈亮[1] 李喆[1] 黄书鹏[1] 李世荣[1] 

机构地区：[1]第三军医大学西南医院整形美容专科医院,重庆400038 [2]广州军区武汉总医院急诊科,武汉430070

出　　处：《第三军医大学学报》2011年第2期160-163,共4页Journal of Third Military Medical University

摘　　要：目的观察染色体10上缺失的磷酸酶和张力蛋白同源物(phosphatase and tensin homologue deleted on chromosome ten,PTEN)编码蛋白对TGF-β1刺激下瘢痕成纤维细胞转分化的抑制作用及其机制。方法用重组PTEN腺病毒转染体外培养的人瘢痕成纤维细胞,倒置荧光显微镜检测PTEN转染组及空载体转染组绿色荧光蛋白表达;将实验分为对照组、PTEN转染组、空载体转染组、TGF-β1刺激组(给予TGF-β1刺激)、PTEN+TGF-β1组(PTEN基因转染后给予TGF-β1刺激)、空载体+TGF-β1组(空载体转染后给予TGF-β1刺激),RT-PCR和Western blot分别检测对照组、PTEN转染组、空载体转染组中PTEN mRNA和蛋白表达;Western blot检测各组中α-SMA、Akt、p-Akt表达水平。结果 PTEN基因转染瘢痕成纤维细胞36h后可见明显绿色荧光蛋白表达,PTEN转染组PTEN mRNA及蛋白的表达明显升高,与对照组及空载体转染组相比,差异有统计学意义(P<0.05),而p-Akt和α-SMA的表达明显下降(P<0.05);TGF-β1组α-SMA表达较对照组明显升高,同时伴有p-Akt表达上升,与对照组、PTEN转染组、PTEN+TGF-β1组相比,差异均有统计学意义(P<0.01);空载体转染组与对照组相比,p-Akt、α-SMA蛋白表达差异均无统计学意义(P>0.05);空载体+TGF-β1组与TGF-β1刺激组相比,p-Akt、α-SMA蛋白表达差异均无统计学意义(P>0.05)。各组细胞的Akt表达差异均无统计学意义(P>0.05)。结论 TGF-β1可促进PI3K/Akt通路活化,使瘢痕成纤维细胞α-SMA表达升高,促进其转分化;空载体转染对p-Akt、α-SMA的表达无明显影响,高表达PTEN可使瘢痕成纤维细胞α-SMA的表达明显降低,并且可拮抗TGF-β1对瘢痕成纤维细胞的转分化作用,其机制可能是抑制PI3K/Akt通路活化。Objective To investigate the inhibitory effects of overexpression of phosphatase and tensin homologue deleted on chromosome ten（PTEN） on the trans-differentiation of hypertrophic scar fibroblast induced by TGF-β1,and the signaling transduction mechanism.Methods Hypertrophic scar fibroblasts were transfected with GFP-PTEN via adenovirus.The cells were divided into 6 groups,normal group,Ad-PTEN group,Ad-Laz group,TGF-β1 stimulation group（10 ng/ml）,Ad-PTEN ＋TGF-β1 group,and empty vector ＋TGF-β1 group.The efficiency of transfection was detected by fluorescence microscopy.The expression of PTEN protein and mRNA in the transfected cells was detected by Western blot analysis and RT-PCR respectively.The expressions of α-SMA,Akt and p-Akt were detected by Western blot analysis.Results Most cells transfected with Ad-PTEN expressed GFP.The expression of PTEN protein and mRNA were strongly increased when the cells were transfected with Ad-PTEN（all P0.05）.In Ad-PTEN group,the expression levels of p-Akt and α-SMA were lower than those in normal group and Ad-Laz group（P0.05）.In TGF-β1 group,the expression levels of α-SMA and p-Akt were higher than those in normal group,Ad-PTEN group and PTEN＋TGF-β1 group（P0.01）.The expression levels of p-Akt and α-SMA had no diversity in normal group and Ad-Laz group（P0.05）,and in Ad-Laz＋TGF-β1group and TGF-β1 group（P0.05）.The expression of Akt was similar in the 6 groups.Conclusion TGF-β1 can promote the expression of α-SMA by activating the PI3K/Akt signal pathway.PTEN inhibits the trans-differentiation of hypertrophic scar fibroblasts induced by TGF-β1 through suppressing the activation of PI3K/Akt signal pathway.

关 键 词：PTEN 成纤维细胞 细胞转分化 PI3K/AKT 

分 类 号：R341[医药卫生—基础医学] R364.31