红芪多糖对糖尿病周围神经病变ob/ob小鼠高迁移率族蛋白1-Toll样受体4信号通路的影响

Effect of Hedysarum polybotrys polysacchcaide on high mobility group box protein1-Toll-like receptor 4 signaling pathway in ob/ob mice with diabetic peripheral neuropathy

目的研究红芪多糖(HPS)对糖尿病周围神经病变(DPN)小鼠外周神经组织中高迁移率族蛋白1-Toll样受体4(HMGB1-TLR4)信号通路的影响。方法将50只10周龄雄性ob/ob小鼠和10只非转基因雄性db/m正常组小鼠随机分为6组,高、中、低剂量实验组分别给予200,100,50 mg·kg^(-1)·d^(-1)红芪多糖,对照组给予硫辛酸30 mg·kg^(-1)·d^(-1),模型组和正常组均给予蒸馏水5m L·kg^(-1)·d^(-1),连续灌胃干预8周。检测各组小鼠的血糖水平;用电生理仪测定坐骨神经传导速度;以Western blot、聚合酶链式反应(PCR)法检测坐骨神经组织HMGB1、TLR4、核因子转录因子κB(NF-κB)、白细胞介素(IL)-1β蛋白和mRNA的表达。结果治疗8周后,正常组、模型组和中、高2个剂量实验组的HMGB1蛋白表达分别为0. 18±0. 03,0. 84±0. 04,0. 57±0. 04和0. 37±0. 08,模型组与正常组比较,差异有统计学意义(P <0. 01),中、高2个剂量实验组与模型组比较,差异均有统计学意义(P <0. 01或P <0. 05)。正常组、模型组和中、高2个剂量实验组的TLR4蛋白表达分别为0. 22±0. 01,1. 05±0. 05,0. 91±0. 04和0. 38±0. 07,模型组与正常组比较,差异有统计学意义(P <0. 01),中、高2个剂量实验组与模型组比较,差异均有统计学意义(P <0. 01或P <0. 05)。基因表达与其相应的蛋白表达趋势一致。结论红芪多糖可能通过调控HMGB1-TLR4信号通路改善糖尿病周围神经病变。

Objective To observe the effect of Hedysarum polybotrys polysacchcaide(HPS)on the signaling pathway of high mobility group box protein 1-toll-like receptor 4(HMGB1-TLR4)in nervous tissue in diabetes mice with diabetic peripheral neuropathy(DPN).Methods A total of 50 ten-week-old male ob/ob mice and 10 non-transgenic male db/m normal mice of the same age were randomly divided into6 groups.Normal group and model group were given 5 m L·kg-1·d-1 distilled water;experimental-L,experimental-M,experimental-H groups were given 50,100,200 mg·kg-1·d-1 HPS;control group was given 30 mg·kg-1·d-1 thioctanate,continuous gavage intervention for8 week.The blood glucose in each group was detected.At the end of treatment,the motor nerve conduction velocity of sciatic nerve was measured by PowerLab.The Western blot,polymerase chain reaction(PCR)were used to measure the protein and mRNA expressions of HMGB1,TLR4,nuclear transcription factor-κB(NF-κB)and interleukin-1β(IL-1β)in sciatic nerve tissue.Results Eight weeks after treatment,the expressions of HGMB1 protein in normal group,model group and experimental-M,experimental-H groups were 0.18±0.03,0.84±0.04,0.57±0.04 and 0.37±0.08,compared with normal group,the HMGB1 protein level in model group had statistical difference(P<0.01);compared with model group,the HMGB1 protein levels in experimental-M and experimental-H groups had statistical difference(P<0.01,P<0.05);the expressions of TLR4 protein in normal group,model group and experimental-M,experimental-H groups were 0.22±0.01,1.05±0.05,0.91±0.04 and0.38±0.07,compared with normal group,the TLR4 protein level in model group had statistical difference(P<0.01);compared with model group,the TLR4 protein levels in experimental-M and experimental-H groups were significantly different(P<0.01,P<0.05).The expression trenf of mRNA is consistent with that of its corresponding protein.Conclusion HPS can reduce the development of nervous injure by controlling inflammatory reaction through the HMGB1-TLR4 signaling pathway.