基于NOD2/NF-κB通路研究雷公藤多苷对糖尿病肾病大鼠的影响

Effect of Tripterygium Wilfordii Polyglycoside on Diabetic Nephropathy Rats Based on NOD2/NF-κB Pathway

目的:基于核苷酸结合寡聚化结构域蛋白2(nucleotide-binding oligomerization domain 2,NOD2)/核转录因子-κB(nuclear transcription factor-κB,NF-κB)通路探讨雷公藤多苷对糖尿病肾病(diabetic nephropathy,DN)大鼠的影响。方法:从42只SPF级雄性SD大鼠中随机选择8只作为空白组,其余大鼠采用高脂高糖饲料喂养联合腹腔注射链脲佐菌素(55 mg·kg^(-1))复制糖尿病肾病模型,将24只造模成功的大鼠随机分为模型组、缬沙坦对照组(8.33 mg·kg^(-1))和雷公藤多苷组(5 mg·kg^(-1)),每组8只,给予相应药物灌胃干预6周,每天1次,空白组与模型组灌胃给予等体积生理盐水。双缩脲法检测大鼠24 h尿蛋白定量(24 hours urinary total protein,24hUTP);全自动生化分析仪测定各组大鼠血糖(blood glucose,GLU)、血清肌酐(serum creatinine,SCr)、血尿素氮(blood urea nitrogen,BUN)水平;HE染色观察肾组织病理学变化;ELISA检测血清NOD2、NF-κB水平;Western Blot检测肾组织NOD2、NF-κB蛋白表达水平;荧光定量PCR检测肾组织NOD2 mRNA、NF-κB mRNA表达水平。结果:与空白组比较,模型组大鼠GLU、SCr、BUN、24hUTP水平明显升高(P<0.05),血清NOD2、NF-κB水平及肾组织NOD2、NF-κB蛋白及mRNA表达水平明显升高(P<0.05);与模型组比较,雷公藤多苷组大鼠GLU、SCr、BUN、24hUTP水平明显降低(P<0.05),血清NOD2、NF-κB水平及肾组织NOD2、NF-κB蛋白及mRNA表达水平明显降低(P<0.05)。HE染色显示:模型组大鼠肾小球肥大,肾小管上皮细胞气球样变性,胞质空泡化,肾血管淤血、扩张,基底膜增厚;与模型组比较,雷公藤多苷组大鼠病理改变较轻,肾小球未见明显病理改变,表现为肾小管轻度增厚、扩张。结论:雷公藤多苷能够有效改善DN大鼠肾损伤,其机制可能与抑制NOD2/NF-κB信号通路有关。

Objective:To explore the effect of tripterygium wilfordii polyglycoside on diabeticnephropathy(DN)rats based on the nucleotide-binding oligomerization domain 2(NOD2)/nuclear transcription factor-κB(NF-κB)pathway.Methods:Eight SPF male SD rats were randomly selected as the blank group,and the rest rats were fed with high-fat and high-sugar diet combined with intraperitoneal injection of streptozotocin(55mg/kg)to establish diabetic nephropathy model.Twenty-four rats successfully established the model were randomly divided into model group,valsartan control group(8.33 mg·kg^(-1))and tripterygium wilfordii polyglycoside group(5 mg·kg^(-1)),with 8 rats in each group,and given corresponding drugs.24-hour urinary protein quantification(24-hour UTP)was detected by biuret method.The levels of bloodglucose(GLU),serumcreatinine(SCr)and bloodureanitrogen(BUN)were measured by automatic biochemical analyzer.HE staining was used to observe the pathological changes of renal tissue.Serum levels of NOD2 and NF-κB were detected by ELISA.WesternBlot was used to detect the expression level of NOD2 and NF-κB protein in renal tissue.The expression levels of NOD2mRNA and NF-κBmRNA in renal tissue were detected by fluorescence quantitative PCR.Results:Compared with the blank group,the levels of GLU,SCr,BUN and 24-hour UTP in the model group were significantly increased(P<0.05),and the levels of NOD2 and NF-κB in serum and the expression of NOD2,NF-κB protein and mRNA in renal tissue were significantly increased(P<0.05).Compared with the model group,the levels of GLU,SCr,BUN and 24-hour UTP in the tripterygium wilfordii polyglycoside group were significantly decreased(P<0.05),and the levels of NOD2 and NF-κB in serum and the expression of NOD2,NF-κB protein and mRNA in renal tissue were significantly decreased(P<0.05).HE staining showed that in the model group,glomerular hypertrophy,balloon degeneration of renal tubular epithelial cells,vacuolation of cytoplasm,congestion and expansion of renal blood vessels and thickening of basement membrane were observed.Compared with the model group,the pathological changes of rats in the tripterygium wilfordii polyglycoside group were mild,and there was no obvious pathological changes in glomerulus,which showed mild thickening and expansion of renal tubules.Conclusion:Tripterygium wilfordii polyglycoside can effectively improve renal injury in DN rats,and its mechanism may be related to the inhibition of NOD2/NF-κB signaling pathway.