糖络宁对高糖环境下大鼠背根神经节神经元细胞氧化应激及PI3K/AKT信号通路的影响  

Effects of Tangluoning on oxidative stress and PI3K/AKT signaling pathway in dorsal root ganglion neurons of rats under high glucose environment

作　　者：李娜 王利莹 李逸潇 刘琴 陈枫 张涛静[2] LI Na;WANG Li-ying;LI Yi-xiao;LIU Qin;CHEN Feng;ZHANG Tao-jing(The Second Clinical Medical College of Beijing University of Chinese Medicine,Beijing 100029,China;Department of Endocrinology,Dongfang Hospital,Beijing University of Chinese Medicine,Beijing 100078)

机构地区：[1]北京中医药大学第二临床医学院,北京100029 [2]北京中医药大学东方医院内分泌科,北京100078

出　　处：《北京中医药》2020年第11期1161-1165,共5页Beijing Journal of Traditional Chinese Medicine

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

摘　　要：目的探讨糖络宁对高糖环境下大鼠背根神经节神经元(DRGn)氧化应激及PI3K/AKT信号通路表达的影响。方法将清洁级6~8周SD雄性大鼠60只随机分为高、中、低浓度糖络宁组及正常组,分别按糖络宁组方生药5、2.5、1.25 g/(kg·d)及等体积蒸馏水灌胃,用于制备高、中、低浓度糖络宁含药血清及正常对照血清。用于制备细胞悬液的背根神经节取自SD胎鼠。将DRGn细胞接种于6孔板,分为正常组、高糖组、高浓度糖络宁组、中浓度糖络宁组、低浓度糖络宁组5组。培养24h后,运用荧光探针技术检测DRGn细胞中ROS水平,酶标仪检测大鼠DRGn细胞SOD、MDA含量,免疫荧光法+免疫印迹法检测PI3K/AKT蛋白表达及其磷酸化水平。结果与正常组相比,高糖组细胞ROS、MDA明显升高(P<0.01),SOD活性明显降低,PI3K、p-PI3K、p-AKT蛋白表达及p-PI3K/PI3K、p-AKT/AKT比值显著降低(P<0.01);高、中、低浓度糖络宁组对上述指标均有显著改善作用,以中浓度糖络宁组为最佳。结论糖络宁可以改善高糖环境下DRGn细胞的氧化应激水平,其机制可能与激活PI3K/AKT通路有关。Objective To studythe effects of Tangluoning(TLN)on oxidative stress and expression of PI3 K/AKT pathway in dorsal root ganglion neurons(DRGn) under high glucose environment.Methods Sixty male SD rats of 6~8 weeks old of clean grade were randomly divided into high,medium and low-concentrations of TLN groups and normal group, andthe crude drugs selected according to the TLN group prescription were given respectively 5,2.5,1.25 g/(kg·d)and equal volume of distilled water by gavage for the preparation of comparative serum.The dorsal root ganglion(DRG) used to prepare cell suspension was obtained from SD fetal rats.DRGn cells were inoculated into 6-well plate and randomly divided into 5 groups: normal group,high glucose group,high concentration of TLN group,medium concentration of TLN group,low concentration of TLN group.After 24 hours of culture,the level of ROS in DRGn cells was detected by fluorescence probe technology,the content of SOD and MDA in rat dorsal root neurons was detected by enzyme-linked immunosorbent assay;PI3 K/Akt protein expression and its phosphate were detected by immunofluorescence and Western blotting Level.Results Compared with the normal group,ROS and MDA were significantly increased(P<0.01),SOD activity significantly decreased,protein expression of PI3 K,p-PI3 K,p-AKT,and p-PI3 K/PI3 K,p-AKT/AKT ratio significantly decreased in high glucose group(P<0.01);the above indexes could be significantly improved in the high,medium and low concentration of TLN groups,and the best was in the medium concentration of TLN group.Conclusion TLN can improve the oxidative stress level of DRGn cells under high glucose environment,and the mechanism may be related to the activation of PI3 K/AKT pathway.

关 键 词：糖尿病周围神经病变 糖络宁 氧化应激 PI3K/AKT信号通路 大鼠 

分 类 号：R285.5[医药卫生—中药学]