益气活血通络方对糖尿病周围神经病变小鼠JNK信号通路蛋白及炎性因子的影响

目的研究益气活血通络方对糖尿病周围神经病变模型小鼠(db/db小鼠)JNK信号通路蛋白及炎性因子的影响,探讨其对db/db小鼠周围神经的保护作用机制。方法 40只db/db小鼠随机分为5组:模型组、硫辛酸组和益气活血通络方高、中、低剂量(6.4、3.2、1.6 g/kg)组,另设8只db/m小鼠作为正常组。灌胃给药,1次/d,连续8周。末次给药后,测定小鼠足底热敏反应时间和运动神经传导速度(MNCV);酶联免疫吸附法(ELISA法)检测血清中白细胞介素-1β(IL-1β)、肿瘤坏死因子-α(TNF-α)水平;Western blot法检测背根神经节中c-jun氨基末端激酶(JNK)和磷酸化的c-jun氨基末端激酶(p-JNK)的蛋白表达。结果与正常组比较,模型组小鼠MNCV减慢,足底热敏反应时间延长,血清中IL-1β、TNF-α水平升高,背根神经节中JNK、p-JNK蛋白的表达增强,差异具有统计学意义(P0.01或P0.05)。与模型组比较,益气活血通络方能提高小鼠MNCV,缩短足底热敏反应时间,降低血清中IL-1β、TNF-α水平,一定程度下调背根神经节中JNK、p-JNK的蛋白表达(P0.01或P0.05)。结论益气活血通络方能抑制JNK信号转导通路的异常激活,减轻db/db小鼠炎症反应,从而保护周围神经的功能。

Influence of Yiqi Huoxue Tongluo Fang on JNK signal pathway and inflammatory factors in mice with diabetic peripheral neuropathy

ObjectiveTo study the influence of Yiqi Huoxue Tongluo Fang (Qi-Replenishing Blood-Activating Collateral-freeing Decoction) on signal pathway of c-jun N-terminal kinase (JNK) and inflammatory factors in mice with diabetic peripheral neuropathy (db/db mice),and discuss its mechanism in protecting peripheral nerve.Methods The db/db mice (n =40) were randomly divided into 5 groups:model group,lipoic acid group,high-dose Yiqi Huoxue Tongluo Fang group (6.4 g/kg,high-dose group),mid-dose Yiqi Huoxue Tongluo Fang group (3.2 g/kg,mid-dose group) and low-dose Yiqi Huoxue Tongluo Fang group (1.6 g/kg,low-dose group),and other db/m mice (n =8) were chosen into normal group.All groups were intragastrically given corresponding drugs once a day for 8 weeks.After the last administration of drugs,the plantar thermal response time and motor nerve conduction velocity (MNCV) were detected.The levels of interleukin-1β (IL-1β) and tumor necrosis factor-α (TNF-α) were detected by using enzyme-linked immunosorbent assay (ELISA).The protein expressions of JNK and phosphorylase JNK (p-JNK) in dorsal root ganglion (DRG) were detected by using Western blot method.Results Compared with normal group,MNCV was slower,plantar thermal response time was longer,levels of IL-1β and TNF-α increased,and protein expressions of JNK and pJNK are stronger in model group (P ＜ 0.01 or P ＜ 0.05).Compared with model group,Yiqi Huoxue Tongluo Fang promoted MNCV,shortened plantar thermal response time,decreased levels of IL-1 β and TNF-α and down-regulated protein expressions of JNK and p-JNK to some extent (P ＜ 0.01 or P ＜0.05).ConclusionYiqi Huoxue Tongluo Fang can inhibit the abnormal activation of JNK signal pathway and relieve inflammatory reaction to protect peripheral nerve in db/db mice.