TGF-β_1对人输尿管平滑肌细胞增殖和迁移及α-SMA表达的影响  

Effects of TGF-β_1 on the Proliferation,Migration and α-SMA Expression of Human Ureteral Smooth Muscle Cells

作　　者：徐桂彬[1,2] 余涵[3] 陈丽[4] 罗亚楠[4] 马猛[4] 王林杰[4] 何永忠[1,2] 彭晔[5] 李逊[1,2] 朱晓琴[4] 

机构地区：[1]广州医科大学附属第五医院泌尿外科,广州510700 [2]广州医科大学微创新技术与产品转化中心,广州510700 [3]湖北医药学院附属襄阳医院,襄阳市第一人民医院病理科,襄阳441000 [4]广州医科大学基础学院,广州511436 [5]广州医科大学附属口腔医院,广州510140

出　　处：《华中科技大学学报（医学版）》2015年第6期656-660,共5页Acta Medicinae Universitatis Scientiae et Technologiae Huazhong

基　　金：广州市卫生局医药卫生科技项目(No.20131A011172);广东省科技计划项目(No.2014A020212344);广东省医学科研基金项目(No.A2014330);2014年广东省教育厅青年创新人才类项目(自然科学类)(No.124);广州市教育系统创新学术团队(No.13C10);广州医科大学博士启动项目(No.2013C60)

摘　　要：目的探讨外源性转化生长因子-β_1(transforming growth factor-β_1,TGF-β_1)对人输尿管平滑肌细胞(USMCs)生物学功能的影响。方法取正常的输尿管平滑肌细胞分为4组:1对照组;2TGF-β_1组(10μg/L TGF-β_1处理);3TGF-β_1拮抗剂组(30μmol/L SB-431542处理);4TGF-β_1+TGF-β_1拮抗剂组(10μg/L TGF-β_1+30μmol/L SB-431542处理)。MTT法检测各组输尿管平滑肌细胞在处理0、24、48、72h的增殖情况;细胞划痕实验检测各组输尿管平滑肌细胞处理24h、48h时的迁移能力;各组细胞处理48h后分别应用RT-qPCR和Western blot检测α-平滑肌肌动蛋白(α-SMA)的表达变化。结果MTT实验显示与对照组相比,TGF-β_1处理24h后USMCs增殖率增加(P<0.05),处理48h、72h后增殖最明显(P<0.01),拮抗剂组细胞生长明显受抑制(P<0.05或P<0.01),TGF-β_1联合SB-431542共同处理细胞后其增殖活性与对照组相比无明显差异;细胞划痕实验显示TGF-β_1组与对照组相比,各时间点细胞迁移距离明显增加(P<0.01),拮抗剂组24h和48h细胞迁移距离与对照组相比明显减小(P<0.01),TGF-β_1联合拮抗剂处理组迁移距离与对照组相比差异无统计学意义;RT-qPCR和Western blot结果显示:与对照组相比,经TGF-β_1处理组α-SMA mRNA和蛋白的表达明显增加(均P<0.01),TGF-β_1拮抗剂则明显下调α-SMA mRNA和蛋白的表达(P<0.01)。结论 TGF-β_1能够明显上调输尿管平滑肌细胞中α-SMA mRNA和蛋白的表达,并能增强细胞增殖和迁移的生物活性,拮抗TGF-β_1信号路径可明显对α-SMA的表达及细胞增殖和迁移产生抑制效应。Objective To explore the effect of TGF-β_1 on the biological functions of ureteral smooth muscle cells（USMCs）.Methods Normal ureteral smooth muscle cells are divided into four groups：1control group;2TGF-β_1group（10μg/L TGF-β_1）;3TGF-β_1antagonist group（30μmol/L SB-431542）;4TGF-β_1＋ TGF-β_1antagonist group（10μg/L TGF-β_1＋ 30μmol/L SB-431542）.MTT assay was used to detect the proliferation of USMCs after treatment for 0,24,48 and 72h,and cell wound-healing assay to observe the migration ability of USMCs at 24 and 48h.The expression change ofα-smooth muscleactin at 48 hwas measured by RT-qPCR and Western blotthing,respectively.Results MTT assay showed that USMCs proliferation rate was increased after 24hTGF-β_1treatment when compared with control group（P〈0.05）.It was further increased at 48 and 72h（P〈0.01）.In contrast,the proliferation in TGF-β_1antagonist group was significantly decreased（P〈0.05 or P〈0.01）.There was no significant difference in the proliferation rate between the TGF-β_1＋ TGF-β_1antagonist group and control group（P〈0.05）.Cell wound-healing assay showed that the cell migration distance was significantly increased at each time point in TGF-β_1group when compared with the control group（P〈0.01）,which was significantly inhibited in TGF-β_1antagonist group（P〈0.01）at 24 and 48h.The migration distance in TGF-β_1＋ TGF-β_1antagonist group was not significantly different from that in control group（P〈0.05）.RT-qPCR and Western blotting results showed that theα-SMA mRNA and protein expression levels were significantly increased in TGF-β_1 group as compared with the control group（P〈0.01）,which were down-regulated after treatment with TGF-β_1antagonist SB-431542（P〈0.01）.Conclusion TGF-β_1can obviously increase theα-SMA mRNA and protein expression in USMCs,and enhance the proliferation and migration of USMCs.Inhibition of TGF-β_1signaling pathway can significantly suppress the expression ofα-SMA,and the prolif

关 键 词：输尿管平滑肌细胞 转化生长因子β_1 Α-平滑肌肌动蛋白 增殖 迁移 

分 类 号：R691.2[医药卫生—泌尿科学]