活血祛痰法对糖尿病肾病大鼠核因子活化B细胞κ轻链增强子NF-κB信号通路的影响

Effect of Promoting Blood Circulation and Eliminating Phlegm on NF-κB Signal Pathway in Diabetic Nephropathy Rat

目的观察活血祛痰法对糖尿病肾病大鼠核转录因子-κB(NF-κB)信号传导通路的影响,探讨其对DN防治作用及机制。方法建立糖尿病肾病(DN)大鼠模型,随机分为正常组、模型组、中药组,观察各组大鼠24 h饮水量、24 h尿蛋白量、血肌酐、空腹血糖水平等指标;病理切片过碘酸雪夫(PAS)染色观察肾脏的病理改变;采用一步法定量PCR检测肾组织NF-κB基因的表达;免疫印迹法(Western blot)检测P65蛋白质含量。结果治疗后,模型组尿蛋白量较正常组呈上身趋势,中药组治疗后,24 h尿蛋白量、血肌酐明显低于模型组(P<0.01);模型组NF-κB基因在肾脏中表达量明显高于正常组,治疗后,中药组NF-κB基因在肾脏中表达水平较模型组明显下调(P<0.01);相对于模型组P65蛋白表达上调,中药组治疗后P65蛋白表达量下降,接近正常组。结论活血祛痰法可用于治疗早期糖尿病肾病,机制可能与其能抑制DN大鼠肾脏中NF-κB信号传导通路的激活有关。

Objective To observe the effect of activating blood circulation and eliminating phlegm method on NF - KB signal transduc- tion pathway in diabetic nephropathy rats, to explore the therapeutic effect and mechanism of DN. Methods To establish the rat model of DN, rats were randomly divided into normal group, model group, Chinese medicine group, the index of 24h drinking water and 24h urine protein, serum creatinine, fasting blood glucose level were observed; the pathological changes of kidney were observed by pathological section PAS staining; the expression of NF - KB gene in renal tissue was detected by one step quantitative PCR; P65 protein content was detected by Western blot technology. Results After the treatment, the model group compared with normal group, the urine protein quantity was in upper trend ;24h urinary protein and serum creatinine of traditional Chinese medicine group after treatment were significantly lower than that of mod- el group （P 〈0.01 ） ; the model group of NF - KB expression in the kidney was significantly higher than that of normal group, after treat- ment, traditional Chinese medicine group of NF - KB gene expression level in the kidney was significantly lower than model group （ P 〈 0. 01 ） ; the expression of P65 protein was up - regulated in model group but decreased in traditional Chinese medicine group after treatment, and close to the normal group. Conclusion Activating blood circulation and eliminating phlegm method can be used for the treatment of early diabetic nephropathy, the mechanism may be related to the inhibition of the DN rat kidney NF - KB signal transduction pathway activation.