糖络宁通过调控miR-146a减轻高糖诱导的大鼠背根神经节细胞炎症反应机制研究  

Study on the mechanism of Tangluoning to reduce the inflammatory response of rat dorsal root ganglion cells induced by high glucose with regulating Mir-146a

作　　者：李矜 张晨[1] 辛竞妍 吴长江 朱雨菲 张涛静[2] LI Jin;ZHANG Chen;XIN Jingyan;WU Changjiang;ZHU Yufei;ZHANG Taojing(The Second Clinical Medical College of Beijing University of Chinese Medicine,Beijing 100029,China)

机构地区：[1]北京中医药大学第二临床医院,100029 [2]北京中医药大学东方医院内分泌科

出　　处：《环球中医药》2022年第11期2059-2063,共5页Global Traditional Chinese Medicine

基　　金：国家自然科学基金面上项目(81774223,81473663);2019年度北京中医药大学校级杰出青年人才项目(BUCM-2019-JCRC010)。

摘　　要：目的 探讨糖络宁通过调控miR-146a减轻高糖诱导下的大鼠背根神经节细胞炎症反应的机制。方法 清洁级SD大鼠20只随机均分为两组,以糖络宁生药及等体积蒸馏水灌胃以制备糖络宁含药血清及正常血清。以新生胎鼠背根神经节细胞作为研究对象,采用高糖刺激构建细胞损伤模型,分为正常组(空白血清培养)、高糖组(150 mmol/L葡萄糖+空白血清培养)、糖络宁组(150 mmol/L葡萄糖+糖络宁含药血清培养)、miR-146a基因过表达组(150 mmol/L葡萄糖+miR-146a激动剂)4个组,48小时后,收集各组细胞并进行相关检测。细胞增殖毒性检测法检测各组细胞24、48及72小时活力,酶联免疫吸附法检测各组细胞上清液中炎症因子(白介素-6、白介素-1β、肿瘤坏死因子-α)的蛋白表达情况。逆转录聚合酶链反应法检测各组细胞miR-146a和炎症因子的表达情况。结果 (1)与正常组相比,高糖组、糖络宁组及miR-146a基因过表达组光密度值均明显下降(P<0.05),炎症因子蛋白及基因表达水平均明显升高(P<0.05),高糖组、糖络宁组miR-146a基因表达水平明显下降(P<0.05)。(2)与高糖组相比,糖络宁组与miR-146a基因过表达组细胞光密度值均明显升高(P<0.05),炎症因子蛋白及基因表达水平均明显下降(P<0.05),miR-146a基因表达水平明显升高(P<0.05)。结论 糖络宁能够明显减轻高糖诱导的背根神经节细胞炎症反应,其部分机制可能是通过上调miR-146a的表达水平实现的。Objective To investigate the mechanism of Tangluoning in reducing the inflammatory response of rat dorsal root ganglion cells induced by high glucose with regulating Mir-146a. Methods Rat dorsal root ganglion cells were used as research objects to construct cell damage model by high glucose stimulation. Four groups were divided into the normal group(blank serum culture), the high glucose group(150 mmol/L glucose+blank serum culture), the Tangluoning group(150 mmol/L glucose+Tangluoning drug-containing serum culture) and the Mir-146a gene overexpression group(150 mmol/L glucose + Mir-146a agonist). 48 hours later, cells in each group were collected and tested. The cell viability in each group was detected by CCK-8 method, the protein expression of inflammatory factors(IL-6, IL-1β and TNF-α) in supernatant of each group was detected by enzyme-linked immunoassay(ELISA), and the expression of Mir-146a and inflammatory factors(IL-6, IL-1β and TNF-α) were detected by RT-PCR. Results Compared with the normal group, OD value in the high-glucose group, the Tangluoning group and the Mir-146a gene overexpression group were significantly decreased(P<0.05), the expression levels of inflammatory factor protein and gene were significantly increased(P<0.05), mir-146a gene expression level of the high-glucose group and Tangluoning group was significantly decreased(P<0.05). Compared with the high glucose group, the OD value of cells in the Tangluoning group and the Mir-146a gene overexpression group was significantly increased(P<0.05), the expression levels of inflammatory factor protein and gene were significantly decreased(P<0.05), and the expression level of Mir-146a gene was significantly increased(P<0.05). Conclusion Tangluoning can significantly reduce the inflammatory response of dorsal root ganglion cells induced by high glucose, partly due to up-regulating the expression level of Mir-146a.

关 键 词：糖尿病周围神经病变 糖络宁 炎症因子 大鼠背根神经节 炎症反应 

分 类 号：R285.5[医药卫生—中药学]