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。
Abstract：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-α and 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 the expression of inflammatory and fibrotic components in mesangial cells induced by high glucose by up-regulating the expression of SOCS2.
李晓艳, 莫立稳, 程莹, 吴绮楠. 白果内酯调控细胞因子信号转导抑制因子2抑制高糖诱导的肾小球系膜细胞中炎症与纤维化成分的表达[J]. 河北医学, 2022, 28(12): 2007-2012.
LI Xiaoyan, MO Liwen, CHENG Ying, et al. Bilobalide Regulates SOCS2 to Inhibit the Expression of Inflammatory Fibrotic Components in High Glucose-Induced Glomerular Mesangial Cells. HeBei Med, 2022, 28(12): 2007-2012.
[1] Giralt-Lopez A,Molina-Van den Bosch M,Vergara A,et al.Revisiting experimental models of diabetic nephropathy[J].Int Mol Sci,2020,21(10):3587-3609. [2] 余俊键,陈瑜,杨升平.白果内酯对大鼠心肌抗氧化机制的初步研究[J].医学信息,2019,32(19):83-85. [3] 王蕾,张雄,陈赵乐,等.白果内酯对糖氧剥夺小鼠原代海马神经元内钙离子浓度的影响[J].神经解剖学杂志,2020,36(1):40-44. [4] 董志超,高琦,毛建娜.白果内酯通过TLR4/TAK1/NF-κB通路对非酒精性脂肪肝炎大鼠模型肝纤维化的调控作用[J].沈阳药科大学学报,2019,36(8):716-722. [5] 杜艺,崔彤霞,马文洁,等.荞麦黄酮通过下调lncRNA DLX6-AS1抑制高糖诱导的肾小球系膜细胞炎症因子表达和纤维化[J].中国中西医结合肾病杂志,2021,22(1):37-40. [6] 黄茸茸,陆松侠,许燕,等.白果内酯对Aβ25-35诱导HT22细胞损伤及NLRP-1炎症小体激活的影响[J].中成药,2020,42(9):2317-2323. [7] Wang X,Liu Y,Rong J,et al.LncRNA HCP5 knockdown inhibits high glucose-induced excessive proliferation,fibrosis and inflammation of human glomerular mesangial cells by regulating the miR-93-5p/HMGA2 axis[J].BMC Endocr Disord,2021,21(1):134-148. [8] Yoon JJ,Park JH,Lee YJ,et al.Protective effects of ethanolic extract from rhizome of polygoni avicularis against renal fibrosis and inflammation in a diabetic nephropathy model[J].Int Mol Sci,2021,22(13):7230-7245. [9] Xiang J,Yang F,Zhu W,et al.Bilobalide inhibits inflammation and promotes the expression of Aβ degrading enzymes in astrocytes to rescue neuronal deficiency in AD models[J].Transl Psychiatry,2021,11(1):542-549. [10] Min XQ,Xie Y.LncRNA CASC2 alleviates the progression of diabetic nephropathy by regulating the miR-144/SOCS2 signalling axis[J].Kidney Blood Press Res,2020,45(6):837-849.
2022, Vol. 28 
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