当归多糖对DPN大鼠TLR4/MyD88/NF-κB通路抑制影响

doi:10.12092/j.issn.1009-2501.2018.12.004

摘要/Abstract

摘要：

目的: 分析当归多糖对糖尿病神经病变（DPN）大鼠TLR4/MyD88/NF-κB通路抑制影响。方法: 选取SPF级Wistar雄性大鼠105只，随机选取15只作为正常组，剩余90只大鼠制备DPN模型，成功造模75只，随机分成5组，模型组、当归多糖低、中、高剂量组和阳性对照组，每组15只。模型组和正常组大鼠采用2 mL/100 g生理盐水灌胃，当归多糖低、中、高剂量组大鼠给予当归多糖药液灌胃，阳性对照组给予弥可保+二甲双胍溶液灌胃，每日给药体积均为2 mL/100 g。每天1次，连续灌胃10 d。给药3、10 d后检测大鼠机械缩足反应阈值（PWT）、热缩足反射潜伏期（PWL），末次给药后检测大鼠空腹血糖、血清丙二醛（MDA）、超氧化物歧化酶（SOD）、过氧化氢酶（CAT）活性、谷胱甘肽过氧化物酶（GSH-Px）、白细胞介素-6（IL-6）、髓鞘碱性蛋白（MBP）、TNF-α及C反应蛋白（CRP）含量，检测大鼠运动神经传导速度（MNCV）、感觉神经传导速度（SNCV），大鼠坐骨神经内核因子-κB（NF-κB） mRNA、髓样分子因子88（MyD88） mRNA及Toll样受体4（TLR4） mRNA表达。结果: 模型组及当归多糖低剂量组大鼠空腹血糖、MDA含量显著高于正常组，CAT、SOD及GSH-Px含量显著低于正常组，当归多糖中、高剂量组大鼠空腹血糖、MDA含量显著低于模型组、阳性对照组，CAT、SOD及GSH-Px含量显著高于模型组、阳性对照组，差异有统计学意义（P＜0.05）；模型组、当归多糖低剂量组大鼠血清IL-6、MBP、TNF-α及CRP含量均显著高于正常组，当归多糖中、高剂量组大鼠血清IL-6、MBP、TNF-α及CRP含量均显著低于模型组、阳性对照组，差异有统计学意义（P＜0.05）；模型组、当归多糖低剂量组大鼠NF-κB mRNA、MyD88 mRNA及TLR4 mRNA表达量均显著高于正常组，当归多糖中、高剂量组大鼠NF-κB mRNA、MyD88 mRNA及TLR4 mRNA表达量均显著低于模型组、阳性对照组，差异有统计学意义（P＜0.05）。结论: 当归多糖可显著改善DPN大鼠的周围神经损伤状态，促进神经传导的恢复，其机制可能与阻断TLR4/MyD88/NF-κB信号路径的转导，降低炎性因子水平，减轻神经的氧化应激损伤有关。

关键词: 糖尿病周围神经病变, Toll样受体4, 髓样分子因子88, 核转录因子-κB, 当归多糖

Abstract:

AIM: To analyze the effects of angelica sinensis polysaccharides on TLR4/MyD88/NF-κB signaling pathway in the inhibition of diabetic neuropathy (DPN) in rats. METHODS: A total of 105 SPF-level Wistar male rats were randomly selected and 15 were selected as the normal group, and the remaining 90 rats were used to prepare the DPN model. Seventy-five models were successfully established and were randomly divided into 5 groups, i.e. the model group, the angelica polysaccharide low, medium and high dose groups and positive control group, with 15 rats in each group. The rats in the model group and the normal group were intragastrically administered with 2 mL/100 g normal saline. The rats in the low, middle and high doses of angelica polysaccharide were given the polysaccharide of angelica sinensis polysaccharide, and the positive control group was given the Mi Kebao + metformin solution. The administration volume was 2 mL/100 g once a day for a continuous gavage of 10 days. After 3 and 10 days of administration, the mechanical contraction threshold (PWT) and heat-shrinking reflex latency (PWL) were measured. After the last administration, the rats were tested for fasting blood glucose, serum malondialdehyde (MDA) and superoxide dismutase. (SOD), catalase (CAT) activity, glutathione peroxidase (GSH-Px), interleukin-6 (IL-6), myelin basic protein (MBP), TNF-α And C-reactive protein (CRP) content, detecting motor motor conduction velocity (MNCV), sensory nerve conduction velocity (SNCV), rat sciatic nerve factor-κB (NF-κB) mRNA, myeloid molecular factor 88 (MyD88) mRNA and Toll-like receptor 4 (TLR4) mRNA expression. RESULTS: The fasting blood glucose and MDA content in the model group and the low dose group of angelica polysaccharide were significantly higher than those in the normal group. The contents of CAT, SOD and GSH-Px were significantly lower than those in the normal group. The fasting blood glucose and MDA content in the middle and high dose groups of angelica polysaccharide were significant. The levels of CAT, SOD and GSH-Px in the model group and the positive control group were significantly higher than those in the model group and the positive control group, and the difference was statistically significant (P＜0.05). The levels of serum IL-6, MBP, TNF-α and CRP in the model group and angelica polysaccharide low dose group were significantly higher than those in the normal group, and the serum IL-6, MBP and TNF-α in the middle and high dose groups of angelica sinensis polysaccharide. The CRP content was significantly lower than that of the model group and the positive control group, and the difference was statistically significant (P＜0.05).The expressions of NF-κB mRNA, MyD88 mRNA and TLR4 mRNA in the model group and the low dose group of angelica sinensis polysaccharide were significantly higher than those in the normal group, and the expression of NF-κB mRNA, MyD88 mRNA and TLR4 mRNA in the middle and high dose groups of angelica sinensis polysaccharide. Both were significantly lower than the model group and the positive control group, and the difference was statistically significant (P＜0.05). CONCLUSION: Angelica sinensis polysaccharides can significantly reduce the serum inflammatory factors and oxidative stress response in DPN rats. The mechanism may be that it inhibits the expression of TLR4, which affects the downstream gene expression of MyD88 signaling pathway, which is related to the inhibition of NF-κB activation.

Key words: diabetic peripheral neuropathy, toll-like receptor 4, myeloid molecule factor 88, nuclear transcription factor-κB, angelica sinensis polysaccharide

中图分类号:

周夏慧，王庆来，朱雪梅，潘蓉蓉，潘胜莲，朱方会，王丁，罗晓红. 当归多糖对DPN大鼠TLR4/MyD88/NF-κB通路抑制影响[J]. 中国临床药理学与治疗学, 2018, 23(12): 1340-1347.

ZHOU Xiahui, WANG Qinglai, ZHU Xuemei, PAN Rongrong, PAN Shenglian, ZHU Fanghui, WANG Ding, LUO Xiaohong. Effects of angelica sinensis polysaccharide on TLR4/MyD88/NF-κB pathway in rats with diabetic neuropathy[J]. Chinese Journal of Clinical Pharmacology and Therapeutics, 2018, 23(12): 1340-1347.

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