[1]郭亮,毕胜,王珍祥,等.PTEN抑制增生性瘢痕成纤维细胞转分化作用及其机制[J].陆军军医大学学报(原第三军医大学学报),2011,33(02):160-163.
 Guo Liang,Bi Sheng,Wang Zhenxiang,et al.PTEN inhibits trans-differentiation of hypertrophic scar fibroblasts in vitro[J].J Amry Med Univ (J Third Mil Med Univ),2011,33(02):160-163.
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PTEN抑制增生性瘢痕成纤维细胞转分化作用及其机制(/HTML )
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陆军军医大学学报(原第三军医大学学报)[ISSN:1000-5404/CN:51-1095/R]

卷:
33卷
期数:
2011年第02期
页码:
160-163
栏目:
论著
出版日期:
2011-01-30

文章信息/Info

Title:
PTEN inhibits trans-differentiation of hypertrophic scar fibroblasts in vitro
作者:
郭亮毕胜王珍祥陈亮李喆黄书鹏李世荣
第三军医大学西南医院整形美容专科医院;广州军区武汉总医院急诊科
Author(s):
Guo Liang Bi Sheng Wang Zhenxiang Chen Liang Li Zhe Huang Shupeng Li Shirong
Department of Plastic and Reconstructive Surgery, Southwest Hospital, Third Military Medical University, Chongqing, 400038; Department of Emergency, Wuhan General Hospital of Guangzhou Military Command, Wuhan, Hubei Province, 430070, China
关键词:
PTEN成纤维细胞细胞转分化PI3K/Akt
Keywords:
phosphatase and tensin homologue deleted on chromosome ten fibroblast trans-differentiation PI3K/ Akt
分类号:
R341;R364.31;R394.2
文献标志码:
A
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36 h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。
Abstract:
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 P<0.05). In Ad-PTEN group, the expression levels of p-Akt and α-SMA were lower than those in normal group and Ad-Laz group(P<0.05). 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 (P<0.01). The expression levels of p-Akt and α-SMA had no diversity in normal group and Ad-Laz group (P>0.05), and in Ad-Laz+TGF-β1group and TGF-β1 group  (P>0.05). 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.

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更新日期/Last Update: 2011-01-19