摘要
目的:研究白果内酯是否通过调控细胞因子信号转导抑制因子2(SOCS2)影响高糖诱导的肾小球系膜细胞中炎症与纤维化成分的表达。方法:人肾小球系膜细胞(HMC)进行如下分组:NG组(正常培养)、HG组(30mmoL/L葡萄糖)、HG+低、中、高白果内酯组(6μmoL/L、12μmoL/L、24μmoL/L白果内酯和30mmoL/L葡萄糖)、HG+高白果内酯+NC组(转染小干扰RNA阴性对照、24μmoL/L白果内酯和30mmoL/L葡萄糖)和HG+高白果内酯+干扰SOCS2组(转染针对SOCS2的小干扰RNA、24μmoL/L白果内酯和30mmoL/L葡萄糖)。利用Western blotting检测SOCS2和纤维化成分转化生长因子-β1(TGF-β1)、结缔组织生长因子(CTGF)、纤连蛋白(FN)的蛋白表达,酶联免疫吸附测定法(ELISA)检测炎性因子肿瘤坏死因子-α(TNF-α)与白细胞介素-6(IL-6)表达。结果:与NG组相比,HG组肾小球系膜HMC细胞中TGF-β1[(0.82±0.04)vs(0.23±0.02)]、CTGF[(0.69±0.05)vs(0.12±0.01)]、FN的蛋白表达量[(0.81±0.06)vs(0.19±0.03)]、TNF-α[(580.58±41.78)vs(148.46±11.47)]pg/mL、IL-6表达水平[(427.82±31.51)vs(60.72±6.24)]pg/mL均升高,而SOCS2的蛋白表达量[(0.22±0.02)vs(0.79±0.06)]减少(t=39.578,33.536,27.727,29.921,34.285,27.037,P<0.05);与HG组相比,HG+低、中、高白果内酯组肾小球系膜HMC细胞中TGF-β1、CTGF、FN的蛋白表达量、TNF-α、IL-6表达水平逐渐降低,但SOCS2的蛋白表达量逐渐增加(P<0.05),并呈现浓度依赖性。与HG+高白果内酯+NC组相比,HG+高白果内酯+干扰SOCS2组肾小球系膜HMC细胞的SOCS2蛋白表达量减少,TGF-β1、CTGF、FN蛋白表达量和TNF-α、IL-6表达水平均提升(P<0.05)。结论:白果内酯通过上调SOCS2的表达,抑制高糖诱导的肾小球系膜细胞中炎症及纤维化成分的表达。
Objective:To investigate whether bilobalide affects the expression of inflammatory and fibrotic components in high glucose-induced glomerular mesangial cells by regulating suppressor of cytokine signaling 2(SOCS2).Methods:Human mesangial cells(HMC)were divided into the following groups:NG group(normal culture),HG group(30 mmoL/L glucose),HG+low,medium and high bilobalide groups(6μmoL/L,12μmoL/L,24μmoL/L bilobalide and 30 mmoL/L glucose),HG+high bilobalide+NC group(transfected small interfering RNA negative control,24μmoL/L bilobalide and 30 mmoL/L glucose)and HG+high bilobalide+interfering SOCS2 group(transfected with small interfering RNA against SOCS2,24μmoL/L bilobalide and 30 mmoL/L glucose).Western blotting was used to detect the protein expressions of SOCS2 and fibrotic components transforming growth factor-β1(TGF-β1),connective tissue growth factor(CTGF),and fibronectin(FN).Enzyme-linked immunosorbent assay(ELISA)was used to detect inflammation factors tumor necrosis factor-α(TNF-α)and interleukin-6(IL-6)expression.Results:Compared with the NG group,the protein expressions of TGF-β1[(0.82±0.04)vs(0.23±0.02)],CTGF[(0.69±0.05)vs(0.12±0.01)],FN[(0.81±0.06)vs(0.19±0.03)],TNF-α[(580.58±41.78)vs(148.46±11.47)]pg/mL and IL-6[(427.82±31.51)vs(60.72±6.24)]pg/mL in the glomerular HMC cells of the HG group were increased,while the protein expression of SOCS2[(0.22±0.02)vs(0.79±0.06)]was decreased(P<0.05);compared with the HG group,the protein expressions of TGF-β1,CTGF and FN,and the expression levels of TNF-αand IL-6 in the glomerular mesangial HMC cells of the HG+low,medium and high bilobalide groups gradually decreased,but the protein expression of SOCS2 gradually increased(P<0.05),and all in a concentration-dependent manner.Compared with HG+high bilobalide+NC group,the expression of SOCS2 protein in glomerular mesangial HMC cells in HG+high bilobalide+interfering SOCS2 group decreased,the expression of TGF-β1,CTGF,FN and TNF-α,IL-6 was increased(P<0.05).Conclusion:Bilobalide can inhibit
作者
李晓艳
莫立稳
程莹
吴绮楠
LI Xiaoyan;MO Liwen;CHENG Ying(General Hospital of Western Theater,Sichuan Chengdu 610031,China)
出处
《河北医学》
CAS
2022年第12期2007-2012,共6页
Hebei Medicine
基金
四川省干部保健科研项目普及应用项目,(编号:川干研2022-1302)。
