熊果酸对瘢痕成纤维细胞生物学功能及TGF-β1/Smads通路的影响  

Effects of Ursolic Acid on the Biological Function of Scar Fibroblasts and the TGF-β1/Smads Pathway

作　　者：李大鹏 刘菲菲 董利焕 王梦楠 靳楷 胡太平 王璞 LI Dapeng;LIU Feifei;DONG Lihuan;WANG Mengnan;JIN Kai;HU Taiping;WANG Pu(Department of Medical Cosmetology,Handan Central Hospital,Handan 056000,China)

机构地区：[1]邯郸市中心医院,医学美容科,邯郸056000

出　　处：《中国细胞生物学学报》2024年第11期1927-1935,共9页Chinese Journal of Cell Biology

摘　　要：该文探讨了熊果酸对瘢痕成纤维细胞生物学功能及转化生长因子β1(TGF-β1)/Smads通路的影响。取对数生长期的增生性瘢痕成纤维细胞(HSFB),用不同剂量熊果酸(0、10、20、30、40、50、60μmol/L)干预HSFB后,MTT法确定细胞存活率;将HSFB分为对照组(0μmol/L熊果酸)、L熊果酸组(20μmol/L熊果酸)、M-熊果酸组(40μmol/L熊果酸)、H-熊果酸组(60μmol/L熊果酸)、H熊果酸+SRI-011381组(60μmol/L熊果酸+10μmol/L TGF-β1/Smads通路激活剂SRI-011381)。流式细胞术和TUNEL染色测定细胞凋亡情况;Transwell实验测定细胞迁移和侵袭能力;Western blot测定各组中TGF-β1/Smads通路蛋白(TGF-β1、p-Smad2、p-Smad3、Smad4)、纤维化蛋白(Col 1A1、Col 3A1、α-SMA)及凋亡蛋白(Bax、Bcl-2)的表达情况。结果显示,随着熊果酸浓度的增加,HSFB存活率逐渐降低,且呈剂量依赖性(P<0.05)。与0μmol/L熊果酸对比,10、20、30、40、50、60μmol/L熊果酸处理后细胞存活率降低,差异显著(P<0.05)。与对照组对比,L-熊果酸组、M-熊果酸组和H熊果酸组的HSFB凋亡率、TUNEL阳性细胞比例、Bax蛋白表达水平均升高(P<0.05),迁移细胞数量、侵袭细胞数量以及纤维化标记蛋白(Col 1A1、Col 3A1、α-SMA)、Bcl-2、TGF-β1/Smads通路相关蛋白表达水平均降低(P<0.05),且熊果酸高剂量相比于低剂量变化程度更显著(P<0.05);与H熊果酸组对比,H-熊果酸+SRI-011381组的HSFB凋亡率、TUNEL阳性细胞比例、Bax蛋白表达水平均降低(P<0.05),迁移细胞数量、侵袭细胞数量以及纤维化标记蛋白(Col 1A1、Col 3A1、α-SMA)、Bcl-2、TGF-β1/Smads通路相关蛋白表达水平均升高(P<0.05)。由此提示,熊果酸可有效抑制HSFB的增殖、侵袭、迁移和纤维化,诱导细胞凋亡,其机制可能与抑制TGF-β1/Smads信号通路激活有关。The aim of this study is to investigate the effects of ursolic acid on the biological function of scar fibroblasts and the TGF-β1(transforming growth factor-β1)/Smads pathway.By culture with different doses of ursolic acid(0,10,20,30,40,50,60μmol/L),cell survival rate of HSFB(hypertrophic scar fibroblast)in logarithmic growth phase was firstly detected by MTT methods.After that,five experimental groups,named as control group(without ursolic acid adding in culture medium),L-ursolic acid group(with 20μmol/L ursolic acid adding in culture medium),M-ursolic acid group(with 40μmol/L ursolic acid adding in culture medium),H-ursolic acid group(with 60μmol/L ursolic acid adding in culture medium),H-ursolic acid+SRI-011381 group(with 60μmol/L ursolic acid and 10μmol/L TGF-β1/Smads pathway activator SRI-011381),were selected for the further study.Apoptosis of HSFB when culture in the above groups was determined by flow cytometry and TUNEL staining.Transwell experiment was applied to measure migration and invasion abilities of HSFB.Western blot was applied to determine the expression of TGF-β1/Smads pathway proteins(TGF-β1,p-Smad2,p-Smad3,Smad4),fibrotic proteins(Col 1A1,Col 3A1,α-SMA),and apoptotic proteins(Bax,Bcl-2)in each group.The result showed that with the concentration of ursolic acid increased in culture medium,the survival rate of HSFB gradually decreased in a dosedependent manner(P<0.05).As compared with the blank control,the apoptosis rate,proportion of TUNEL positive cells and expression of Bax protein in HSFB in the L-ursolic acid group,M-ursolic acid group,and H-ursolic acid group were all increased(P<0.05),while the numbers of migrating cells,invading cells,and the expression of fibrotic marker proteins(Col 1A1,Col 3A1,α-SMA,Bcl-2,TGF-β1/Smads pathway related proteins)of HSFB were all reduced on the contrary(P<0.05).Those degree of change in H-ursolic acid group was more obvious than that of L-ursolic acid group(P<0.05).Furthermore,the apoptosis rate,proportion of TUNEL positive cells and expre

关 键 词：增生性瘢痕 成纤维细胞 转化生长因子β1(TGF-β1)/Smads 生物学功能 

分 类 号：R622[医药卫生—整形外科]