沉默单核细胞趋化蛋白3对人增生性瘢痕成纤维细胞增殖、迁移及凋亡的影响  

Effects of silencing MCP-3 on proliferation,migration and apoptosis of fibroblasts from human hypertrophic scar

作　　者：田阳 王飞[1] 徐骁然 王德华[1] 徐虎 李庭超 TIAN Yang;WANG Fei;XU Xiao-ran;WANG De-hua;XU Hu;LI Ting-chao(Department of Burn and plastic surgery,the First People's Hospital of Zunyi,Zunyi563000,China)

机构地区：[1]遵义市第一人民医院烧伤整形外科,贵州遵义563000 [2]遵义市第一人民医院检验科,贵州遵义563000 [3]遵义市第一人民医院病理科,贵州遵义563000

出　　处：《临床皮肤科杂志》2022年第9期518-523,共6页Journal of Clinical Dermatology

摘　　要：目的:探讨沉默单核细胞趋化蛋白3(MCP-3)对人增生性瘢痕成纤维细胞增殖、迁移及凋亡等功能的影响。方法:取增生性瘢痕组织和正常皮肤组织,qRT-PCR检测正常皮肤组织和增生性瘢痕组织中MCP-3 mRNA表达水平;分离培养增生性瘢痕组织成纤维细胞(HSF)和正常皮肤组织成纤维细胞(NSF),qRT-PCR和Western blot检测两种成纤维细胞中MCP-3 mRNA和蛋白表达水平。采用MCP-3干扰慢病毒(shRNA-MCP-3)和其阴性对照病毒(shRNA-NC)感染HSF,MTT检测各组细胞增殖活性,流式细胞术检测各组细胞凋亡情况,Transwell实验检测细胞的迁移能力,qRT-PCR检测各组细胞中MCP-3、collagenⅠ和collagenⅢmRNA水平,Western blot检测细胞中凋亡相关蛋白MCP-3、Bax、Bcl-2、Cleaved-caspase-3及collagenⅠ和collagenⅢ蛋白表达水平。结果:MCP-3在增生性瘢痕组织及HSF中高表达。与blank组和shRNA-NC组比较,shRNA-MCP-3组细胞中MCP-3 mRNA和蛋白表达水平显著下降(P<0.01),细胞增殖活性、迁移能力及细胞中collagenⅠ、collagenⅢ和Bcl-2表达水平均显著降低(P<0.01),同时细胞凋亡率及凋亡蛋白Bax和Cleaved-caspase-3表达水平显著上升(P<0.01)。结论:干扰MCP-3表达可能通过抑制细胞增殖和诱导细胞凋亡,减少细胞合成胶原,从而发挥抑制瘢痕形成与发展的作用。Objective: To investigate the effects of monocyte chemotactic protein 3(MCP-3) on proliferation, migration and apoptosis of fibroblasts from human hypertrophic scar. Methods: Hypertrophic scar tissue and normal skin tissue were collected. Hypertrophic scar fibroblasts(HSF) and normal skin fibroblasts(NSF) were isolated and cultured. The levels of MCP-3mRNA expression was detected by using qRT-PCR, and the levels of MCP-3 protein were detected by using Western blot.MCP-3 interference lentivirus(shRNA-MCP-3) and its negative control lentivirus(shRNA-NC) were used to infect cultured HSF. The MTT assay was used to measure cell proliferation, flow cytometry was used to study cell apoptosis, and Transwell assay was used to measure cell migration ability, in both HSF and NSF. The mRNA expression levels of MCP-3, collagenⅠand collagenⅢ were detected by using qRT-PCR. The protein expression levels of MCP-3, Bax, Bcl 2, Cleaved-caspase-3,collagenⅠ and collagenⅢ were detected using Western blot. Results: MCP-3 was highly expressed in hypertrophic scar tissue and HSF. Compared with the blank group and shRNA-NC group, the levels of MCP-3 mRNA and protein expression in shRNAMCP-3 group were significantly decreased(P<0.01). The cell proliferation, migration ability and expression levels of collagen Ⅰ、collagenⅢ and Bcl-2 in shRNA-MCP-3 group were also significantly decreased(P<0.01), while apoptosis rate and Bax and Cleaved-caspase-3 expression of apoptotic protein were significantly increased(P<0.01) in shRNA-MCP-3 group, compared with the blank group and shRNA-NC group. Conclusion: Interference with MCP-3 expression may inhibit cell proliferation, induce cell apoptosis, and reduce cell collagen synthesis, which ultimately reduce scar formation.

关 键 词：单核细胞趋化蛋白 增生性瘢痕 迁移 胶原沉积 

分 类 号：R751[医药卫生—皮肤病学与性病学]