摘要
目的研究芒果苷元对TGF-β1诱导的HK-2细胞EMT的影响。方法MTT法检测各浓度芒果苷元对HK-2细胞活力的影响;用TGF-β1诱导HK-2细胞成EMT模型,并给以芒果苷元干预,在TGF-β1诱导24 h时显微镜下拍照记录细胞形态变化,划痕实验法检测细胞迁移能力,Transwell法检测细胞侵袭能力,Western blot法检测纤连蛋白(fibronectin,FN)和Ⅰ型胶原(collagen typeⅠ,ColⅠ)蛋白表达水平。结果(1)芒果苷元(10^(-9)~10^(-5) mol/L)浓度下HK-2细胞活力均无明显变化(P>0.05);(2)与正常组相比,模型组细胞形态变长变梭,迁移和侵袭能力显著增强(P<0.05,0.01),FN和ColⅠ蛋白表达水平显著上调(P<0.05,0.01);与模型组相比,芒果苷元组细胞能维持原来的鹅卵石形,迁移和侵袭能力显著下降(P<0.05,0.01),FN和ColⅠ蛋白表达水平显著下调(P<0.05)。结论芒果苷元能抑制TGF-β1诱导的HK-2细胞迁移和侵袭,阻止细胞形态的改变,下调细胞外基质成分FN和ColⅠ的蛋白表达,从而抑制EMT的发展。
Objective To study the effects of norathyriol on epithelial-mesenchymal transition of HK-2 cells induced by TGF-β1.Methods MTT assay was used to detect the effect of norathyriol on HK-2 cell viability.The HK-2 cells were divided into normal group,model group,norathyriol(10^(-8),10^(-7),10^(-6) mol·L-1)groups and positive control(benzbromarone,10^(-7) mol/L)group.The norathyriol and benzbromarone were incubated for 4 h before HK-2 cells were treated by TGF-β1(10 ng/mL)to induce EMT.Then,the HK-2 cells were cultured for another 24 h or 48 h.Scratch assay was used to detect the migration motility of HK-2 cells.Transwell invasion assay was used to detect invasion ability of HK-2 cells.The proteins expression of fibronectin(FN)and collagen typeⅠ(ColⅠ)were detected by western blot.Results(1)The norathyriol(10^(-9)-10^(-5) mol/L)has no effects on the HK-2 cell viability,compared with the normal group(P>0.05).(2)Compared with the normal group,the cells morphology of the model group changed from the original cobblestone shape to a long fusiform shape,the migration rate and the invasive ability of the model group was significantly increased(P<0.01).The protein expression of Col and FN in the model group was significantly increased.Compared with the model group,the cells morphology of norathyriol(10^(-8),10^(-7),10^(-6) mol/L)and benzbromarone(10^(-7) mol/L)groups maintain thecobblestone shape partly,the migration rate(10^(-8),10^(-7),10^(-6) mol/L)and the invasive ability(10^(-6) mol/L)of norathyriol were significantly decreased(P<0.01);The protein expression of Col and FN were significantly down-regulated in the norathyriol(10^(-6) mol/L)group and the benzbromarone group(P<0.05,0.01).Conclusion Norathyriol can inhibit the migration and invasion of HK-2 cells induced by TGF-β1 and down-regulate the protein expression of fibronectin(FN)and collagen typeⅠ(ColⅠ).
作者
徐爱萍
林华
高丽辉
李玲
陈梦威
王歌
杨娟
牛艳芬
XU Ai-ping;LIN Hua;GAO Li-hui;LI Ling;CHENG Meng-wei;WANG Ge;YANG Juan;NIU Yan-fen(Biomedical Engineering Research Center of Kunming Medical University,Kunming Yunnan 650500,China)
出处
《昆明医科大学学报》
CAS
2021年第7期1-6,共6页
Journal of Kunming Medical University
基金
国家自然科学基金资助项目(81760666)
云南省科技厅-昆明医科大学应用基础研究联合专项基金资助项目(2018FE001(-166))
云南省教育厅科学研究基金资助项目(2019Y0350)。
