连翘苷抑制HMGB1/RAGE信号通路减轻大鼠糖尿病周围神经病变坐骨神经损伤

Phillyrin alleviates sciatic nerve injury in diabetic peripheral neuropathy rats by inhibiting HMGB1/RAGE signaling pathway

目的 基于高迁移率族蛋白Box-1(HMGB1)/晚期糖基化终产物受体(RAGE)信号通路探讨连翘苷(PHI)对糖尿病周围神经病变(DPN)大鼠坐骨神经损伤的影响。方法 利用高脂高糖饮食并注射链脲佐菌素(STZ)建立DPN大鼠模型,随机分为模型组、连翘苷低剂量(PHI-L)(50 mg/kg)组、连翘苷中剂量(PHI-M)(100 mg/kg)组、连翘苷高剂量(PHI-H)(200 mg/kg)组、阳性药物(甲钴胺,250μg/kg)组;另取正常饲料喂养大鼠为对照组。每组10只。采用BL-420S生物机能实验系统测定坐骨神经传导速度;血糖仪检测空腹血糖(FBG)水平;ELISA试剂盒检测血清HbAlc、IL-6、TNF-α水平;电镜观察坐骨神经超微结构形态;试剂盒检测坐骨神经中ROS、SOD、MDA、MBP水平;RT-qPCR、Western blot法检测坐骨神经中HMGB1、RAGE mRNA和蛋白水平。结果 与模型组比较,PHI-M组、PHI-H组、阳性药物组大鼠病理损伤明显减轻,坐骨神经传导速度、MBP、SOD水平显著恢复,FBG、HbAlc、IL-6、TNF-α、ROS、MDA、HMGB1、RAGE的mRNA及蛋白水平显著下降(P均<0.05)。结论 PHI可降低炎症反应,减轻大鼠DPN,发挥治疗作用,其机制可能与抑制HMGB1/RAGE信号通路有关。

This study was designed to investigate the effet of phillyrin(PHI) on sciatic nerve injury in diabetic peripheral neuropathy(DPN) rats based on the high mobility group protein box-1(HMGB1)/advanced glycation end product receptor(RAGE) signal pathway.DPN rat model was established by high fat and high sugar diet combined with injection of streptozotocin(STZ) solution,and the rats were randomly grouped into model group,phillyrin low dose(PHI-L,50 mg/kg) group,phillyrin medium dose(PHI-M,100 mg/kg) group,phillyrin high dose(PHI-H,200mg/kg) group,and positive drug(mecobalamin,250 μg/kg) group,while another rats with normal diet were treated as the control group.Each group consisted of 10 rats.The conduction velocity of sciatic nerve was measured by BL-420S biological function experiment system;fasting blood glucose(FBG) level was detected by blood glucose meter;the levels of serum HbAlc,IL-6 and TNF-α were measured with ELISA kits;the ultrastructure of sciatic nerve was observed with electron microscope;the levels of ROS,SOD,MDA and MBP in sciatic nerve were detected with commercial kits;the mRNA and protein levels of HMGB1 and RAGE in sciatic nerve were detected by RT-qPCR and Western blotting.Compared with model group,pathological injury of rats in PHI-M group,PHI-H group and positive drug group was significantly alleviated,the sciatic nerve conduction velocity and MBP/SOD levels were significantly recovered,the levels of FBG,HbAlc,IL-6,TNF-α,and the mRNA and protein levels of ROS,MDA,HMGB1 and RAGE were decreased significantly(P<0.05).In conclusion,PHI can reduce the inflammatory reaction,reduce DPN in rats,thus plays a therapeutic role,and the mechanism may be related to the inhibition of HMGB1/RAGE signal pathway.