基于PI3K/Akt信号通路探索益气活血祛风通络法治疗糖尿病肾病的作用机制  

Action mechanism of Yiqi Huoxue Qufeng Tongluo method in treating diabetic nephropathy based on PI3K/Akt signaling pathway

作　　者：黄为钧[1] 赵进喜[2] 王世东[2] 傅强[2] 宫晴[2] 张华[2] 吴文静[3] 申子龙[4] HUANG Weijun;ZHAO Jinxi;WANG Shidong(Section II of Endocrinology ＆ Nephropathy Department of Dongzhimen Hospital Affiliated to Beijing University of Chinese Medicine,Beijing 100700,China)

机构地区：[1]北京中医药大学东直门医院重点实验室,100700 [2]北京中医药大学东直门医院肾病内分泌二科 [3]湖北省中医院 [4]首都医科大学北京中医医院

出　　处：《环球中医药》2018年第8期1196-1203,共8页Global Traditional Chinese Medicine

基　　金：国家自然科学基金(81473664)

摘　　要：目的基于PI3K/Akt信号通路探索益气活血祛风通络法治疗糖尿病肾病的分子机制。方法 40只GK大鼠,随机分为模型组、假手术组、益气活血组、祛风通络组、益气活血祛风通络(全方)组,每组8只。糖尿病肾病模型通过高脂高糖喂养加单肾切除术复制。益气活血组、祛风通络组、全方组分别给予益气活血颗粒、祛风通络颗粒、益气活血祛风通络颗粒,模型组、假手术组给予纯净水。干预8周,观察大鼠在干预后肾脏的病理改变以及24小时尿蛋白变化,同时肾脏组织的pPI3K、p-Akt和PTEN的表达情况采用Western blotting以及免疫组化法检测,PI3K mRNA、Akt mRNA以及PTEN mRNA的表达水平采用RT-PCR法检测。结果 HE染色结果显示,中药干预的大鼠肾小管上皮细胞空泡变性程度、纤维化以及肾小球硬化比模型组以及假手术组要轻,且在肾间质纤维化方面,全方组要好于益气活血组以及祛风通络组;PASM染色和Masson染色阳性率均显示益气活血组、祛风通络组、全方组阳性率明显低于模型组(P<0.01),但全方组阳性率与益气活血组以及祛风通络组无统计学差异(P>0.05);益气活血组、祛风通络组、全方组的24小时尿蛋白在中药干预后明显减少,而且与模型组对比具有显著性的差异(P<0.01)。Western blotting结果显示,中药干预的三组pPI3K、p-Akt的表达明显高于模型组(P<0.01),而PTEN的表达显著低于模型组(P<0.01)。免疫组化结果显示,p-PI3K、p-Akt在药物干预的三组表达显著低于模型组(P<0.05);PTEN在祛风通络组以及全方组的表达显著低于模型组(P<0.01),在活血化瘀组表达水平与模型组差异无统计学意义(P>0.05)。RT-PCR结果显示PI3K mRNA、Akt mRNA在中药干预的三组均明显低于模型组(P<0.05),而PTEN mRNA的表达水平则显著高于模型组(P<0.01)。结论益气活血、祛风通络法治疗糖尿病肾病均可以起到降低蛋白尿、减轻肾间质纤维化以Objective To search the molecule mechanism of Yiqi Huoxue Oufeng Tongluo method in treating diabetic nephropathy （DN） based on PI3 K/Akt signaling pathway. Methods A total of 40 GKrats were randomly divided into model group, sham-operated group, Yiqi Huoxue group, Qufeng Tongluo group and Yiqi Huoxue Oufeng Tongluo group. The DN model was established by teeding with high-tat diet and unilateral nephrectomy. Yiqi Huoxue group, Qufeng Tongluo group and giqi Huoxue Qufeng Tongluo group were given Yiqi Huoxue granule, Qufeng Tongluo granule and Yiqi Huoxue Qufeng Tongluo granule, while the model group and sham-operation group were given pure water. The intelarention time was 8 weeks. 24-hour urine protein was detected and the renal pathological changes were observred. The expressions of p-PI3K, p-Akt and PTEN in renal tissues were detected by Western blotting and immunohistochemisn3% while RT-PCR was used to detect the expression of PI3K mRNA, Akt mRNA and PTEN mRNA. Results Results of HE staining showed that vacuolar degeneration of renal tubular epithelial cells, renal interstitial fibrosis and glomerulosclerosis in the three groups were lighter than model group and sham-operation group. Moreover, Yiqi Huoxue Qufeng Tongluo group was better than Yiqi Huoxue group and Qufeng Tongluo group in renal interstitial fibrosis. Results of PASM staining and Masson staining showed that the positive rate of three drug interevention groups were significantly lower than that of model group （ P 〈 0.01 ）. But there was no significant difference between the positive rate of whole group and Yiqi Huoxue and Oufeng Tongluo group. The 24-hour urinary protein in three drug intervention groups were significantly decreased after drug intervention, and the difference was significant compared with model group （ P〈0.01 ）. Results of Western blotting showed that expression levels of p-PI3K and p-Akt in three drug intervention groups were significantly higher than that in the model group （P〈0.01）, while the expression lev

关 键 词：P13K/AKT信号通路 益气活血祛风通络法 糖尿病肾病 

分 类 号：R285.5[医药卫生—中药学]