RUNX3对肝星状细胞活化增殖和迁移能力的影响OA
Effect of RUNX3 on the activation,proliferation,and migration capabilities of hepatic stellate cells
目的 探讨靶向沉默矮小相关转录因子3(RUNX3)对小鼠肝星状细胞(HSCs)的增殖和迁移以及随后的胶原沉积等作用.方法 选取小鼠肝星状细胞系(JS-1),在显微镜下观察并鉴定形态.待细胞完全贴壁后使用5 ng/mL 转化生长因子β1(TGF-β1)作用24 h诱导HSCs活化,并用RUNX3慢病毒感染构建RUNX3沉默模型,实验分为Control组、TGF-β1组、TGF-β1+siRNA-NC组和TGF-β1+siRNA-RUNX3组.Western blot实验检测RUNX3、α-平滑肌肌动蛋白(α-SMA)及Ⅰ型胶原(Collagen I)的蛋白表达变化;细胞免疫荧光实验检测α-SMA、RUNX3在HSCs中的表达定位变化;RT-qPCR检测R U N X 3、α-S M A、Collagen I的mRNA表达变化;EdU染色检测HSCs增殖能力;划痕实验和Transwell实验检测HSCs的迁移能力.结果 与Control组比较,经TGF-β1诱导后,HSCs中RUNX3的表达显著增加(P<0.01).与此同时,纤维化相关指标α-SMA、Collagen I的蛋白及mRNA水平均显著上调(P<0.001),且细胞增殖与迁移能力亦明显增强(P<0.001).而与TGF-β1+siRNA-NC组相比,在TGF-β1+siRNA-RUNX3组中的RUNX3以及α-SMA、Collagen I的蛋白及mRNA水平均明显降低(P<0.05),与此同时,HSCs的增殖和迁移能力也被明显抑制(P<0.01).结论 沉默RUNX3能够抑制HSCs中胶原的沉积及HSCs的增殖和迁移,相反,RUNX3可以促进HSC的增殖和迁移能力,并促进HSC的活化.
Objective To investigate the effects of targeted silencing of Runt-related Transcription Factor 3(RUNX3)on the proliferation and migration of Mouse Hepatic Stellate Cells(HSCs),as well as subsequent colla-gen deposition.Methods Mouse hepatic stellate cell line(JS-1)was selected and then morphologically observed and identified under a microscope.After the cells had fully adhered,they were treated with 5 ng/mL of transform-ing growth factor beta 1(TGF-β1)for 24 hours to induce hepatic stellate cell activation.Furthermore,a RUNX3 si-lencing model was established using RUNX3 lentiviral infection.The experiment was divided into four groups:Control group,TGF-β1 group,TGF-β1+siRNA-NC group,and TGF-β1+siRNA-RUNX3 group.Protein expression changes of RUNX3,alpha-smooth muscle actin(α-SMA),and Alpha 1 type I collagen(Collagen I)were detected using Western blot method.Cellular immunofluorescence assays were employed to investigate the deposition changes of α-SMA and RUNX3 in hepatic stellate cells.RT-qPCR was utilized to examine the mRNA expression changes of RUNX3,α-SMA,and Collagen I.The proliferative capacity of hepatic stellate cells was assessed using Edu staining.The migratory ability of hepatic stellate cells was evaluated through wound healing assays and Tran-swell migration experiments.Results Compared with Control group,a significant elevation in RUNX3 was ob-served in the TGF-β1-induced activated HSCs(P<0.01).Meanwhile,the protein and mRNA levels of fibrosis-related markers and α-SMA and Collagen I were significantly upregulated(P<0.001).Additionally,the prolifera-tion and migration capabilities of HSCs were significantly enhanced(P<0.001).In contrast,when compared to TGF-β1+siRNA-NC group,TGF-β1+siRNA-RUNX3 group exhibited a notable decrease in RUNX3 and other re-lated indicators,such as the protein and mRNA levels of α-SMA and Collagen I(P<0.05).Concurrently,the pro-liferation and migration capabilities of HSCs were significantly inhibited in TGF-β1+siRNA-RUNX3 group(P<0.01).Conclusion Silencing RUNX3 can inhibit the deposition of collagen and the proliferation and migration of hepatic stellate cells.Conversely,RUNX3 promotes the proliferation and migration capabilities of HSCs,thereby facilitating the activation of HSC.
凌辉;汪先晨;尤峻柏;范家好;崔笑;沙纪名;余立权
安徽医科大学第二附属医院 肝胆胰外科,合肥 230601安徽医科大学第二附属医院 胸外科,合肥 230601安徽医科大学第二附属医院 胸外科,合肥 230601安徽医科大学第二附属医院 胸外科,合肥 230601安徽医科大学第二附属医院 肝胆胰外科,合肥 230601安徽医科大学第二附属医院 胸外科,合肥 230601安徽医科大学第二附属医院 肝胆胰外科,合肥 230601
医药卫生
RUNX3肝星状细胞活化增殖迁移肝纤维化胶原沉积
RUNX3hepatic stellate cellsactivateproliferationmigrationhepatic fibrosiscollagen deposi-tion
《安徽医科大学学报》 2026 (2)
277-284,8
安徽省自然科学基金项目(编号:1808085MH270)安徽省高校科研计划项目(编号:2024AH050797)合肥市自然科学基金项目(编号:2021038) Fund programs Natural Science Foundation of Anhui Province(No.1808085MH270)Natural Science Re-search Project of Anhui Educational Committee(No.2024AH050797)Natural Science Foundation of Hefei(No.2021038)
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