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高糖环境下糖络宁对大鼠背根神经节神经元细胞凋亡水平及MAPKs信号通路的影响
朱雨菲1      郭宇鑫1      王利莹2      李矜1      张涛静2
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
Effects of Tangluo Ning on apoptosis and MAPKs signaling pathway of rat dorsal root ganglion neurons in high glucose environment#br#
ZHU Yufei1   GUO Yuxin1   WANG Liying2   LI Jin1   ZHANG Taojing2
1.The Second Clinical Medical College, Beijing University of Chinese Medicine, Beijing   100029, China; 
2.Department of Endocrinology, Dongfang Hospital, Beijing University of Chinese Medicine, Beijing   100078, China
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摘要 目的 研究糖络宁对高糖环境下大鼠背根神经节神经元(DRGn)细胞凋亡及丝裂原激活的蛋白激酶(MAPK)信号通路的影响。 方法 6~8周体重(215±15)g清洁级雄性SD大鼠60只,按照随机数字表法分为高、中、低浓度糖络宁组和正常组,每组15只。各组分别以糖络宁组方生药5、2.5、1.25 g/(kg·d)和等量蒸馏水灌胃,制备含药血清和正常对照血清。新生SD胎鼠制备DRGn细胞,分为正常组(10%正常大鼠血清+Neurobasal培养基),高糖组(正常组基础上+50 mmol/L葡萄糖),糖络宁各浓度组(高糖组基础上+10%低、中、高浓度糖络宁含药血清),培养24 h后检测细胞凋亡及JNK1、p-JNK1、c-Jun、p-c-Jun、Map3k8、p-Map3k8蛋白表达。 结果 高糖组细胞凋亡率、p-JNK1、p-c-Jun、Map3k8、p-Map3k8表达及p-JNK1/JNK1、p-c-Jun/c-Jun、p-Map3k8/Map3k8比值高于正常组,差异有统计学意义(P < 0.05或P < 0.01);糖络宁各浓度组细胞凋亡率、p-JNK1、p-c-Jun、p-Map3k8表达及p-JNK1/JNK1、p-c-Jun/c-Jun、p-Map3k8/Map3k8比值明显低于高糖组,差异有高度统计学意义(P < 0.01);中浓度糖络宁组细胞凋亡率、p-JNK1、p-c-Jun、p-Map3k8表达及p-JNK1/JNK1、p-c-Jun/c-Jun、p-Map3k8/Map3k8比值明显低于高、低浓度糖络宁组,差异有统计学意义(P < 0.01或P < 0.05);中、低浓度糖络宁组c-Jun表达低于高糖组,差异有统计学意义(P < 0.05),糖络宁各浓度组JNK1、Map3k8及c-Jun表达比较,差异无统计学意义(P > 0.05)。 结论 糖络宁可能通过抑制MAPKs信号通路降低细胞凋亡水平。
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朱雨菲1 郭宇鑫1 王利莹2 李矜1 张涛静2
关键词 背根神经节神经元细胞糖尿病周围神经病变糖络宁细胞凋亡    
Abstract:Objective To study the effect of Tangluo Ning on apoptosis and mitogen-activated protein kinases (MAPKs) signaling pathway of rat dorsal root ganglion neurons (DRGn) in high glucose environment. Methods Sixty clean grade male SD rats with body weight of (215±15) g at 6-8 weeks were divided into high, medium, and low concentration glucolonin groups and normal group according to random number table method, with 15 rats in each group. Each group was given 5, 2.5, 1.25 g/(kg·d) crude medicine of Tangluo Ning prescription and equal amount of distilled water intragastric administration to prepare drug-containing serum and normal control serum. DRGn cells were prepared from newborn SD fetal rats and were divided into normal group (10% normal rat serum +Neurobasal medium), high glucose group (based on normal group +50 mmol/L glucose), and Tangluo Ning concentration groups (based on high glucose group +10% low, medium, and high concentration Tangluo Ning drug-containing serum). Cell apoptosis and protein expression of JNK1, p-JNK1, c-Jun, p-c-Jun, Map3k8, and p-Map3k8 were detected after 24 h culture. Results The apoptosis rate, expression of p-JNK1, p-c-Jun, Map3k8, and p-Map3k8 and ratio of p-JNK1/JNK1, p-c-Jun/c-Jun, and p-Map3k8/Map3k8 in high glucose group were higher than those in normal group, the differences were statistically significant (P < 0.05 or P < 0.01). The apoptosis rate, expression of p-JNK1, p-c-Jun, and p-Map3k8 and ratio of p-JNK1/JNK1, p-c-Jun/c-Jun, and p-Map3k8/Map3k8 in Tangluo Ning concentration groups were significantly lower than those in high glucose group, the differences were highly statistically significant (P < 0.01). Apoptosis rate, expression of p-JNK1, p-c-Jun, and p-Map3k8 and ratio of p-JNK1/JNK1, p-c-Jun/c-Jun, and p-Map3k8/Map3k8 in middle concentration Tangluo Ning group were significantly lower than those in high and low concentration Tangluo Ning group, the differences were statistically significant (P < 0.01 or P < 0.05). The expression of c-Jun in the medium and low concentration Tangluo Ning groups was lower than that in the high concentration Tangluo Ning group, and the differences were statistically significant (P < 0.05). There was no significant difference in the expression of JNK1, Map3k8 and c-Jun in the Tangluo Ning concentration groups (P > 0.05). Conclusion Tangluo Ning can reduce apoptosis by inhibiting MAPKs signaling pathway.
Key wordsDorsal root ganglion neurons    Diabetic peripheral neuropathy    Tangluo Ning    Apoptosis
    
基金资助:国家自然科学基金面上项目(81774223、814736 63);
北京中医药大学校级杰出青年人才项目(BUCM-2019-JCRC010)。
通讯作者: 张涛静(1981.10-),男,博士,主任医师;研究方向:糖尿病并发症的中医药防治。   
作者简介: 朱雨菲(1996.2-),女,北京中医药大学第二临床医学院2019级中医内科学专业在读硕士研究生,主要从事内分泌方向研究工作。
引用本文:   
朱雨菲1 郭宇鑫1 王利莹2 李矜1 张涛静2. 高糖环境下糖络宁对大鼠背根神经节神经元细胞凋亡水平及MAPKs信号通路的影响[J]. 中国医药导报, 2022, 19(4): 9-13.
ZHU Yufei1 GUO Yuxin1 WANG Liying2 LI Jin1 ZHANG Taojing2. Effects of Tangluo Ning on apoptosis and MAPKs signaling pathway of rat dorsal root ganglion neurons in high glucose environment#br#. 中国医药导报, 2022, 19(4): 9-13.
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