萝卜硫素通过TGF-β/Smad3信号通路对肾病综合征大鼠系膜细胞增生的影响  

Effect of sulforaphane on mesangial cell proliferation in nephrotic syndrome rats through TGF β / Smad3 signaling pathway

作　　者：杜宗华 毛慧慧 龙青 DU Zonghua;MAO Huihui;LONG Qing(Department of Kidney Disease and Rheumatology,Central Hospital of Enshi Autonomous Prefecture,Enshi 445000,Hubei,China;Hemodialysis Center,Central Hospital of En-shi Autonomous Prefecture,Enshi 445000,Hubei,China)

机构地区：[1]恩施土家族苗族自治州中心医院肾病风湿科,湖北恩施445000 [2]恩施土家族苗族自治州中心医院血液透析中心,湖北恩施445000

出　　处：《西部医学》2020年第1期43-48,共6页Medical Journal of West China

摘　　要：目的探讨萝卜硫素通过转化生长因子(TGF-β)/smad3信号通路对肾病综合征大鼠系膜细胞增生的影响及其作用机制。方法采用2μM TGF-β诱导大鼠肾小球系膜细胞(HBZY-1)增生,将细胞分为对照组、TGF-β刺激组、TGF-β联合20μM萝卜硫素组及TGF-β联合40μM萝卜硫素组。5 mg/kg阿霉素诱导肾病综合征大鼠模型,不同剂量萝卜硫素处理HBZY-1细胞及肾病综合征大鼠,将大鼠分为空白对照组、阿霉素刺激组、阿霉素联合30 mg/kg萝卜硫素组及阿霉素联合60 mg/kg萝卜硫素组,每组各8只;采用CCK8法检测HBZY-1细胞增殖,HE染色分析肾病综合征大鼠肾组织系膜基质沉积,Western Blotting方法检测粘连蛋白(FN)、Ⅳ胶原蛋白(Col4)、TGF-β及p-Smad3蛋白表达水平。结果 TGF-β刺激组能明显诱导HBZY-1细胞增生及HBZY-1细胞FN和Col4蛋白表达,与对照组相比差异有统计学意义(P<0.05);TGF-β联合20、40μM萝卜硫组与TGF-β刺激组相比能明显抑制TGF-β诱导的HBZY-1细胞增殖和明显抑制TGF-β诱导的FN和Col4蛋白表达(P<0.05)。阿霉素诱导肾病综合症大鼠肾脏系膜基质沉积增多(P<0.05),而不同剂量萝卜硫素治疗后,能显著降低肾脏系膜基质沉积(P<0.05)。阿霉素刺激组能明显诱导大鼠肌酐、尿素氮及24小时蛋白尿量水平升高和大鼠肾脏组织FN、Col4、TGF-β、p-Smad3蛋白表达,与对照组相比差异有统计学意义(P<0.05);与阿霉素刺激组相比,不同剂量萝卜硫素能明显降低大鼠肌酐、尿素氮及24小时蛋白尿量水平、降低肾脏组织FN、Col4、TGF-β、p-Smad3蛋白表达水平(P<0.05)。结论萝卜硫素可通过下调TGF-β/Smad3信号传导抑制基质合成相关蛋白表达,进而降低了肾病综合征大鼠肾小球系膜细胞增生。Objective To investigate the effect of sulforaphane on mesangial cell proliferation in nephrotic syndrome rats through transforming growth factor(TG-β)/Smad3 signaling pathway and its mechanism. Methods The proliferation of rat glomerular mesangial cells(HBZY-1) was induced by 2 μ m TGF-β. The cells were divided into control group, TGF β stimulation group, TGF β combined with 20 μ m sulforaphane group and TGF-β combined with 40 μ m sulforaphane group. The rat model of nephrotic syndrome was induced by 5 mg/kg adriamycin. Different doses of sulforaphane were used to treat HBZY-1 cells and nephrotic syndrome rats. The rats were divided into blank control group, adriamycin stimulation group, adriamycin combined with 30 mg/kg sulforaphane group and adriamycin combined with 60 mg/kg sulforaphane group, 8 rats in each group. The proliferation of HBZY-1 cells was detected by CCK8 method, mesangial matrix deposition was analyzed by HE staining, and the expression of FN, col4, TGF-β and P Smad3 were detected by Western blotting. Results TGF-β stimulation group can induce the proliferation of HBZY 1 cells and the expression of FN and col4 protein in HBZY-1 cells, which is statistically significant compared with the blank control group(P<0.05). Compared with TGF-β stimulated group, TGF-β combined with 20 and 40 μ m sulforaphane group could significantly inhibit the proliferation of HBZY-1 cells and the expression of FN and col4 protein induced by TGF-β(P<0.05). Adriamycin induced mesangial matrix deposition in rats with nephrotic syndrome increased(P<0.05), while different doses of sulforaphane could significantly reduce mesangial matrix deposition(P<0.05). Adriamycin stimulation group could significantly induce the increase of creatinine, urea nitrogen and 24-hour proteinuria, and the expression of FN, col4, TGF-β, p-Smad3 protein in kidney tissue of rats, which was statistically significant compared with the blank control group(P<0.05). Compared with the adriamycin group, different doses of sulforaphane cou

关 键 词：萝卜硫素 肾病综合症 肾小球系膜细胞 TGF-β/Smad3信号通路 

分 类 号：R692[医药卫生—泌尿科学]