TSG-6对人病理性瘢痕成纤维细胞增殖与凋亡的影响  

Effects of TSG-6 on the proliferation and apoptosis of human pathological scar fibroblasts

作　　者：张苏文[1] 李小静[1] 陈钊[1] 王晖[1] 

机构地区：[1]安徽医科大学第一附属医院整形外科,合肥230022

出　　处：《安徽医科大学学报》2016年第1期102-105,共4页Acta Universitatis Medicinalis Anhui

基　　金：国家自然科学基金(编号:81272107)

摘　　要：目的探讨肿瘤坏死因子α刺激基因6(TSG-6)对人病理性瘢痕成纤维细胞增殖与凋亡的影响。方法取人病理性瘢痕组织共6例,用组织块培养法培养人病理性瘢痕成纤维细胞,体外培养的第3、4代细胞,用浓度为2μg/ml的重组人TSG-6蛋白处理,在作用48 h后,用免疫组化法检测TSG-6作用前后人病理性瘢痕成纤维细胞增殖细胞核抗原(PCNA)的变化,并计算PCNA阳性细胞率;流式细胞术计算、分析人重组TSG-6蛋白作用前后,人病理性瘢痕成纤维细胞的凋亡率;Western blot法检测细胞凋亡相关蛋白p53、半胱天冬酶3(caspase-3)表达的变化。结果 2μg/ml TSG-6体外干预人病理性瘢痕成纤维细胞48 h情况下,人病理性瘢痕成纤维细胞PCNA的阳性细胞率显著降低,差异有统计学意义(P<0.05);细胞凋亡率增高,差异有统计学意义(P<0.05);Western blot法检测显示细胞凋亡相关蛋白p53、caspase-3表达明显增多。结论 TSG-6在体外能够抑制人病理性瘢痕成纤维细胞的增殖并促进其凋亡,TSG-6对细胞的抑制作用可能与TSG-6抑制PCNA基因表达有关,TSG-6诱导人病理性瘢痕成纤维细胞凋亡的作用可能与TSG-6促进细胞凋亡相关蛋白p53、caspase-3表达有关。Objective To observe the effects of tumor necrosis factor-stimulated gene 6 （TSG-6） on the prolifera- tion and apoptosis of human pathological scar fibroblasts. Methods The samples were harvested from human path- ological scar tissue, a total of 6 cases, and then were cultured to pathological scar fibroblasts with adherence meth- od. Cells used in this study were propagated for 3 -4 times before use. The pathological scar fibroblasts were cul- tured with 2 Ixg/ml of recombinant human TSG-6 protein for 48 h. The effects of TSG-6 on cell proliferation were then assessed by immunohistochemieal method at the indicate time. The rate of apoptosis induced by TSG-6 was de- termined by flow cytometry. Western blot was performed to detect the expression levels of caspase-3 and p53 after TSG-6 was treated. Results After treated with TSG-6 for 48 h, cells expressing proliferating cell nuclear antigen, an indicator of cell proliferation, reduced significantly （ P 〈 0.05 ）. On the other hand, the rate of apoptosis of pathological scar fibroblasts induced by TSG-6 with 2 μg/ml was increased （ P 〈 0. 05 ）. The expression levels of apoptosis related proteins caspase-3 and p53 were significantly increased in TSG-6 treated group. Conclusion TSG-6 may inhibit proliferation and induce apoptosis of human pathological scar fibroblast in vitro. The inhibition of proliferation might be caused by inhibition of PCNA gene expression, and the induction of apoptosis might be asso- ciated with promotion expression of easpase-3 and p53.

关 键 词：肿瘤坏死因子α刺激基因6 病理性瘢痕 成纤维细胞 细胞增殖 细胞凋亡 

分 类 号：R619.6[医药卫生—外科学]