毛蕊花糖苷对糖尿病肾病小鼠肾组织中高迁移率族蛋白1及核因子κB表达的影响

Effects of acteoside on the expressions of high mobility group box 1 and nuclear factor-κB in the renal tissues of diabetic nephropathy mice

目的探讨毛蕊花糖苷对糖尿病肾病小鼠肾组织中高迁移率族蛋白1(high mobility group box 1,HMGB1)及核因子κB(nuclear factor-κB,NF-κB)表达的影响。方法采用20只8周龄健康C57BL/6雄性小鼠,选取其中5只作为正常对照组,余15只单次腹腔注射链脲菌素(150 mg/kg)诱导建立1型糖尿病模型后随机平均分为模型组、毛蕊花糖苷组和厄贝沙坦组。持续给药8周后处死小鼠,收集血清、尿液和肾组织,分别进行生化、病理和相关mRNA与蛋白的检测。体外实验采用肾小管上皮细胞(NRK-52E细胞),将其分为对照组(1 g/L葡萄糖)、高糖组(4.5 g/L葡萄糖)和高糖+毛蕊花糖苷组(4.5 g/L葡萄糖+32μmol/L毛蕊花糖苷),分别于48 h、72 h时间点收获细胞。用实时定量PCR和Western印迹法检测HMGB1和NF-κB mRNA及蛋白的表达。结果与正常对照组比较,模型组血糖、24 h尿蛋白量、血尿素氮、血肌酐均升高(均P<0.05),血清、尿液HMGB1均升高(均P<0.05);病理染色可见肾小管间质炎性细胞浸润,系膜扩增改变;HMGB1和NF-κB的mRNA、蛋白表达均增加(均P<0.05)。与模型组比较,毛蕊花糖苷组24 h尿蛋白量、血尿素氮、血肌酐均下降(均P<0.05),血糖无明显变化(P>0.05),血清、尿液HMGB1均下降(均P<0.05),肾脏组织病变减轻,HMGB1和NF-κB的mRNA、蛋白表达下调(均P<0.05)。NRK-52E细胞中,与对照组比较,高糖组HMGB1、NF-κB mRNA及蛋白水平表达均升高(均P<0.05);毛蕊花糖苷干预后,HMGB1、NF-κB mRNA及蛋白水平表达均下调(均P<0.05)。结论毛蕊花糖苷可改善糖尿病肾病小鼠的肾脏病变,其机制可能与降低HMGB1和NF-κB的表达有关。

Objective To investigate the effect of acteoside on the expressions of high mobility group box 1(HMGB1)and nuclear factor-κB(NF-κB)in the renal tissue of diabetic nephropathy mice.Methods Among 20 healthy 8-week old C57BL/6J mice,5 mice were randomly selected as normal control group,the rest were established as type 1 diabetes mellitus(T1DM)models by a single intraperitoneal injection of streptozocin(STZ,150 mg/kg).T1DM mice were randomly divided into three groups:5 mice without treatment,5 mice treated with acteoside and 5 mice treated with irbesartan.After continuous administration for 8 weeks,serum,urine,and kidney tissue were collected for biochemical,pathological,and related mRNA and protein detection.The renal tubular epithelial cells(NRK-52E cells)were divided into control group(1 g/L glucose),high glucose group(4.5 g/L glucose)and high glucose+acteoside group(4.5 g/L glucose+32μmol/L acteoside).Real-time PCR and Western blotting were used to assess the expressions of HMGB1 and NF-κB after 48 hours and 72 hours culturing.Results Compared with normal control group,blood glucose,24-hour quantitative urinary protein,blood urea nitrogen(BUN),serum creatinine(Scr)and blood and urine HMGB1 were significantly increased in model group(all P<0.05),along with interstitial inflammatory cell infiltration and messangial matrix expantion,and the expressions of HMGB1 and NF-κB were significantly enhanced(all P<0.05).Compared with model group,histopathologic changes were alleviated and the mRNA and protein expression levels of HMGB1 and NF-κB were lower in the acteoside group(all P<0.05),while the blood glucose level was maintained at high level(P>0.05),excluding reduced quantitative 24-hour urinary protein,BUN,Scr,and serum and urine HMGB1(all P<0.05).Compared with control group,the mRNA and protein expressions of HMGB1 and NF-κB were increased in high glucose group of NRK-52E cells(all P<0.05).Compared with high glucose group,the mRNA and protein expressions of HMGB1 and NF-κB in high glucose+acteoside group were down-regulated(all P<0.05).Conclusion Acteoside may alleviate the nephropathy in STZ-induced diabetic nephropathy mice by down-regulating the expressions of HMGB1 and NF-κB.