TGF-β1诱导人Tenon囊成纤维细胞增殖及其机制  

TGF-β1 induces proliferation of human Tenon capsule fibroblast and its mechanism

作　　者：童俊[1] 周宁天[2] 袁志兰[1] 

机构地区：[1]南京医科大学第一附属医院眼科,江苏南京210029 [2]南京医科大学第一附属医院心内科,江苏南京210029

出　　处：《南京医科大学学报（自然科学版）》2014年第4期452-456,477,共6页Journal of Nanjing Medical University(Natural Sciences)

基　　金：国家自然科学基金资助(81271001)

摘　　要：目的：观察TGF-β1是否促进人Tenon囊成纤维细胞（HTFs）增殖并且探讨其作用机制。方法：用TGF-β1（0、0.5、1.0、2.0、5.0、10.0 ng/ml）作用于体外培养的3-6代HTFs 24 h,CCK-8法、EdU染色法检测其增殖;采用Western blot法检测HTFs中RhoE的表达及其下游细胞增殖相关蛋白cyclinD1和增殖细胞核抗原（PCNA）的表达,用慢病毒建立RhoE敲降细胞系。结果：TGF-β1可以剂量依赖性促进HTFs增殖,以5 ng/ml TGF-β1处理后有统计学差异,EdU掺入法检测发现,5 ng/ml TGF-β1组EdU标记率明显高于对照组。5 ng/ml TGF-β1组HTFs中RhoE蛋白的表达平均灰度值明显低于对照组,细胞增殖相关蛋白cyclinD1、PCNA的表达平均灰度值高于对照组。细胞中RhoE沉默后,CCK-8法检测细胞活力较对照组上升,流式细胞术发现S期也升高,cyclinD1、PCNA灰度值也上升。结论：TGF-β1可以通过降低RhoE引起下游周期蛋白cyclinD1的表达升高,诱导人Tenon囊成纤维细胞增殖。Objective：To determine whether TGF-β1 induces the proliferation of Tenon capsule fibroblast （HTFs） and explored its mechanism. Methods：In vitro cultured 3-6 generations of HTFs were treated with various doses of TGF-β 1 （0,0.5,1.0,2.0,5.0 and 10.0 ng/ml） for 24 h. Cell proliferation was detected by the cell counting kit-8 （CCK-8） assay and 5-ethynyl-2-deoxyuridine （EdU） incorporation assay. The expression of RhoE and its downstream cell cycle related protein cyclinD1 and proliferating cell nuclear antigen （PCNA） were detected by Western blot assay. The lentivirns was performed to establish the RhoE knockdown cell lines. Results：The proliferation of the TGF-β1-treated HTFs increased in a dose-dependent manner. Treatment with 5 ng/ml TGF-β1 caused a significant difference. EdU incorporation assay showed that the EdU labeling rate in the 5 ng/ml TGF-β1 group were higher than that in the control group. The average gray value of the expression of RhoE protein in HTFs in the 5 ng/ml TGF-β1 group was lower than that in the control group,while The average gray value of the expressions of cell proliferation protein cyclinD1 and PCNA were higher than those in the control group. After RhoE silenced in the cell,an increase of cell proliferation was detected by CCK-8 assay compared to the control group,flow cytometry showed that S period also increased and the gray value of cell proliferation protein cyclinD1 and PCNA were increased. Conclusion：TGF-β1 increases the expression of downstream cell cycle related protein cyclinD 1 via downregulating RhoE, and induces the proliferation of Tenon capsule fibroblast.

关 键 词：TGF-Β1 成纤维细胞 细胞增殖 青光眼 

分 类 号：R775[医药卫生—眼科]