TGF-β潜态相关肽及其截短体抑制TGF-β1诱导的LX-2细胞纤维化研究  

The study of TGF-βlatency associated peptide and truncated forms in inhibiting TGF-β1-induced hepatic fibrosis in LX-2 cells

作　　者：史嘉翊 宋旭东 邱羽菲 潘宇 曹志琴 吴丹[1] SHI Jia-yi(Mudanjiang Medical University,Mudanjiang 157011,Heilongjiang,China)

机构地区：[1]牡丹江医学院生命科学学院,黑龙江牡丹江157011

出　　处：《牡丹江医学院学报》2024年第4期1-4,39,共5页Journal of Mudanjiang Medical University

基　　金：黑龙江省省属高等学校基本科研业务费科研项目(2018-KYYWFMY-0044)。

摘　　要：目的转化生长因子β(transforming growth factor-β,TGF-β)潜态相关肽(latent assotiated peptide,LAP)及其截短体抑制TGF-β1诱导的LX-2细胞纤维化研究。方法LAP全长命名为LAP-F,LAP截短体命名为LAP-B。用TGF-β1对LX-2处理进行纤维化模型的建立。分组后分别加入LAP-F和LAP-B多肽。control组:在正常情况下进行培养;TGF-β组:加入终浓度10 ng/mL的TGF-β1;TGF-β+LAP-F组:造模后加入终浓度60μg/mL的LAP-F;TGF-β+LAP-B组:造模后加入终浓度60μg/mL的LAP-B多肽。通过Western blot进行纤维化指标FN和α-SMA的蛋白水平检测。细胞免疫荧光实验检测细胞中FN和α-SMA的蛋白水平。结果Western blot结果显示,与control组细胞相比,TGF-β组细胞中纤维化相关蛋白FN的蛋白表达水平明显升高(P<0.001),纤维化相关蛋白α-SMA的蛋白表达也明显升高(P<0.01)。与TGF-β组相比,TGF-β+LAP-B组细胞中FN的蛋白表达明显降低(P<0.01),α-SMA的蛋白表达也明显降低(P<0.01)。与TGF-β组相比,TGF-β+LAP-F组细胞中FN的蛋白表达水平明显降低(P<0.01),α-SMA的蛋白表达水平明显降低(P<0.01)。与TGF-β组相比,TGF-β+LAP-F组细胞中FN和α-SMA蛋白荧光强度更低。免疫荧光的结果显示与control组细胞相比,TGF-β组细胞中FN和α-SMA的蛋白荧光强度更高,这意味着纤维化相关蛋白表达增多。与TGF-β组相比,TGF-β+LAP-B组细胞中FN和α-SMA蛋白荧光强度更低。与TGF-β组相比,TGF-β+LAP-F组细胞中FN和α-SMA蛋白荧光强度更低。结论LAP-F和LAP-B两种多肽都具有抗LX-2细胞纤维化能力,为后续研究提供基础。Objective To investigate the inhibition of TGF-β1-induced LX-2 liver fibrosis by transforming growth factor-β(TGF-β)latent assotiated peptide(LAP)and its truncated forms.Methods The full length of LAP was named LAP-F and the truncated LAP was named LAP-B.The fibrosis model was established by treating LX-2 with TGF-β1.After grouping,LAP-F and LAP-B peptides were added,respectively.Control group:Culture was carried out under normal conditions;TGF-βgroup:TGF-β1 with a final concentration of 10 ng/mL was added;TGF-β+LAP-F group:LAP-F with a final concentration of 60μg/mL was added after modeling;TGF-β+LAP-B group:LAP-B protein with a final concentration of 60μg/mL was added after modeling.The protein levels of fibrosis indicators FN andα-SMA were detected by Western blot and cellular immunofluorescence assay.Results Western blot results showed that the expression level of fibrosis-related protein FN in TGF-βgroup was significantly increased compared with the control group(P<0.001),and the expression level of fibrosis-related proteinα-SMA was also significantly increased(P<0.01).Compared with TGF-βgroup,FN protein expression in TGF-β+LAP-B group was significantly decreased(P<0.01),andα-SMA protein expression was also significantly decreased(P<0.01).Compared with TGF-βgroup,FN protein expression level andα-SMA protein expression level in TGF-β+LAP-F group were significantly decreased(P<0.01).The fluorescence intensity of FN andα-SMA protein in TGF-β+LAP-F group was lower than that in TGF-βgroup.Immunofluorescence results showed that FN andα-SMA protein fluorescence intensity was higher in TGF-βcells than in control cells,indicating increased expression of fibrosis-associated proteins.The fluorescence intensity of FN andα-SMA proteins in TGF-β+LAP-B cells was lower than that in TGF-βgroup.The fluorescence intensity of FN andα-SMA protein in TGF-β+LAP-F group was lower than that in TGF-βgroup.Conclusion Both LAP-F and LAP-B peptides have anti-fibrosis ability of LX-2 cells,which provides the

关 键 词：肝纤维化 TGF-Β LAP LX-2 

分 类 号：R512.62[医药卫生—内科学]