中国组织工程研究

中国组织工程研究 ›› 2023, Vol. 27 ›› Issue (32): 5162-5167.doi: 10.12307/2023.826

• 神经组织构建 nerve tissue construction • 上一篇    下一篇

糖痹康减轻糖尿病周围神经病变大鼠坐骨神经的氧化应激损伤

蒋昇源1，张亚奇1，毕境新2，张程斐3，张秋娥4，穆晓红1，白惠中1，左心玮3，刘铜华2，秦灵灵5，6   

  1. 1北京中医药大学东直门医院，北京市  100700；北京中医药大学，2教育部中医养生学重点实验室，3生命科学学院，4中医学院，5科技处，北京市  100029；6教育部高等学校学科创新引智基地，北京市  100029
  • 收稿日期:2022-09-01 接受日期:2022-11-16 出版日期:2023-11-18 发布日期:2023-03-23
  • 通讯作者: 穆晓红，博士，教授，主任医师，博士生导师，北京中医药大学东直门医院，北京市 100700
  • 作者简介:蒋昇源，男，1993年生，山东省龙口市人，汉族，北京中医药大学在读博士，主要从事中医药促神经修复等研究。
  • 基金资助:
    国家自然科学基金面上项目(81874467)，项目负责人：穆晓红

Tangbikang reduces sciatic nerve oxidative stress injury in a rat model of diabetic peripheral neuropathy

Jiang Shengyuan1, Zhang Yaqi1, Bi Jingxin2, Zhang Chengfei3, Zhang Qiue4, Mu Xiaohong1, Bai Huizhong1, Zuo Xinwei3, Liu Tonghua2, Qin Lingling5, 6   

  1. 1Dongzhimen Hospital, Beijing University of Chinese Medicine, Beijing 100700, China; 2Key Laboratory of Health Preservation of Traditional Chinese Medicine, Ministry of Education, 3College of Life Sciences, 4College of Traditional Chinese Medicine, 5Technology Department, Beijing University of Chinese Medicine, Beijing 100029, China; 6Academic Innovation Introduction Base of the Ministry of Education, Beijing 100029, China
  • Received:2022-09-01 Accepted:2022-11-16 Online:2023-11-18 Published:2023-03-23
  • Contact: Mu Xiaohong, MD, Professor, Chief physician, Doctoral supervisor, Dongzhimen Hospital, Beijing University of Chinese Medicine, Beijing 100700, China
  • About author:Jiang Shengyuan, MD candidate, Dongzhimen Hospital, Beijing University of Chinese Medicine, Beijing 100700, China
  • Supported by:
    the National Natural Science Foundation of China, No. 81874467 (to MXH)

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摘要：


文题释义：
糖痹康：中药复方糖痹康是由传统经方黄芪桂枝五物汤加减优化而得，主要用于糖尿病周围神经病变，现已被证实具有改善血糖及坐骨神经传导速度、调控施万细胞自噬等作用。
氧化应激：线粒体内过量的活性氧累积会诱导细胞器氧化应激，大量氧化中间产物积存于细胞，直接损伤如蛋白质、脂质等各类生物分子，诱导细胞死亡。

背景：中药复方糖痹康具有改善血糖、减轻糖尿病周围神经病变的作用，但其调控作用机制尚未清楚。
目的：观察中药复方糖痹康对糖尿病大鼠坐骨神经氧化应激的影响，探索糖痹康减轻糖尿病周围神经病变的靶向调控机制。
方法：30只SD大鼠随机分为正常组、模型组和糖痹康组，每组10只。后两组构建糖尿病周围神经病变模型，成模后分别灌胃生理盐水或糖痹康2.5 g/(kg·d)，持续12周。给药后0，4，8，12周检测大鼠的体质量、空腹血糖水平；通过热板实验及Von Frey实验检测给药12周各组大鼠热痛阈及机械痛阈；给药12周后取材，取材前检测各组大鼠坐骨神经传导速度及坐骨神经血流速度，苏木精-伊红染色和透射电镜检测观察大鼠坐骨神经形态学变化，采用比色法和ELISA试剂盒分别检测坐骨神经中超氧化物歧化酶、丙二醛、白细胞介素1β、肿瘤坏死因子α表达，Western blot检测坐骨神经组织PI3K/AKT/FoxO1通路相关蛋白表达水平。
结果与结论：①糖痹康组大鼠造模后4，8，12周血糖水平均显著低于模型组(P < 0.05)，模型组大鼠第12周体质量明显低于糖痹康组(P < 0.05)；②造模后12周，糖痹康组机械痛阈及热痛阈相较于模型组有明显改善(P < 0.05)，糖痹康组坐骨神经传导速度及血流速度均明显高于模型组(P < 0.05)；③糖痹康组神经纤维排列秩序、致密程度、髓鞘化程度均优于模型组；④ELISA及比色法结果显示，与模型组相比，糖痹康组坐骨神经组织中丙二醛、白细胞介素1β、肿瘤坏死因子α蛋白表达显著降低，超氧化物歧化酶蛋白表达升高(P < 0.05)；⑤Western blot结果显示，糖痹康组坐骨神经组织中PI3K、p-AKT/AKT、p-FoxO1/FoxO1蛋白表达量较模型组升高(P < 0.05)；⑥糖痹康可改善糖尿病大鼠坐骨神经功能、减轻坐骨神经氧化应激损伤，其机制可能与糖痹康调控PI3K/AKT/FoxO1通路相关。
https://orcid.org/0000-0003-4391-2360(蒋昇源)

中国组织工程研究杂志出版内容重点：组织构建；骨细胞；软骨细胞；细胞培养；成纤维细胞；血管内皮细胞；骨质疏松；组织工程

关键词: 糖痹康, 糖尿病周围神经病变, 线粒体, 氧化应激, PI3K/AKT/FoxO1信号通路

Abstract: BACKGROUND: Tangbikang is a Traditional Chinese Medicine compound that can improve blood glucose and alleviate diabetic peripheral neuropathy. However, its regulatory mechanism is still unclear.
OBJECTIVE: To observe the effect of Tangbikang on oxidative stress of the sciatic nerve in diabetic rats and to explore the targeted regulatory mechanism of Tangbikang in alleviating diabetic peripheral neuropathy. 
METHODS: Thirty Sprague-Dawley rats were randomly divided into normal group, model group and Tangbikang group, with 10 rats in each group. Animal models of diabetic peripheral neuropathy were established in the model group and Tangbikang group. After modeling, normal saline or Tangbikang (2.5 g/kg per day) was administered orally for 12 weeks. Body mass and fasting blood glucose levels of rats were detected at 0, 4, 8 and 12 weeks after administration. Hot plate test and Von Frey test were used to detect the thermal and mechanical pain thresholds of rats at 12 weeks after administration. Sciatic nerve conduction velocity and blood flow velocity were detected before sampling. Hematoxylin-eosin staining and transmission electron microscopy were used to observe the morphological changes of the sciatic nerve. The expression levels of superoxide dismutase, malondialdehyde, interleukin 1β, and tumor necrosis factor α in the sciatic nerve were detected by colorimetric method and ELISA kit. The expression levels of PI3K/AKT/FoxO1 pathway related proteins in the sciatic nerve tissue were detected by western blot assay.
RESULTS AND CONCLUSION: The fasting blood glucose level of rats in the Tangbikang group was significantly lower than that in the model group at 4, 8, and 12 weeks after modeling (P < 0.05), and the body mass of rats in the model group was significantly lower than that in the Tangbikang group (P < 0.05). Compared with the model group, the mechanical pain threshold and thermal pain threshold were significantly improved in the Tangbikang group 12 weeks after modeling (P < 0.05), and the sciatic nerve conduction velocity and blood flow velocity in the Tangbikang group were significantly higher than those in the model group 
(P < 0.05). The order, density and myelination of nerve fibers in the Tangbikang group were better than those in the model group. ELISA and colorimetric results showed that compared with the model group, the levels of malondialdehyde, interleukin 1β, and tumor necrosis factor α proteins in the sciatic nerve tissue were significantly decreased in the Tangbikang group, while the level of superoxide dismutase protein was significantly increased (P < 0.05). Western blot results showed that the expressions of PI3K, p-Akt/AKT, and p-FoxO1/FoxO1 proteins in the sciatic nerve of the Tangbikang group were significantly higher than those of the model group (P < 0.05). To conclude, Tangbikang improves sciatic nerve function and reduces sciatic nerve oxidative stress injury in diabetic rats, and the mechanism may be related to the regulation of PI3K/AKT/FoxO1 pathway by Tangbikang.

Key words: Tangbikang, diabetic peripheral neuropathy, mitochondria, oxidative stress, PI3K/AKT/FoxO1 signaling pathway

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