通络糖泰方对糖尿病周围神经病变大鼠JNK信号转导通路的影响

Effect of Tongluo Tangtai Power on JNK Signaling Pathway in Rats of Diabetic Peripheral Neuropathy

目的:观察通络糖泰方对糖尿病周围神经病变(DPN)大鼠JNK信号转导通路的影响,并探讨其可能的作用机制。方法:SPF级GK大鼠给予高脂饲料喂养建立DPN大鼠模型,50只造模成功的GK大鼠随机分为5组,每组大鼠均为10只,分别为模型组,通络糖泰方高、中、低剂量组,西药联合(二甲双胍+弥可保)组,同时设立正常组Wistar大鼠为10只,分别给予相应处理4周后以免疫组化方法检测坐骨神经组织磷酸化氨基末端激酶(p-JNK),半胱氨酸天冬氨酸蛋白酶-3(Caspase-3)蛋白的表达水平,实时荧光定量聚合酶链式反应(Real-time PCR)方法检测JNK,胰岛素样生长因子1(IGF-1)mRNA的表达水平,酶联免疫吸附法检测血清肿瘤坏死因子-α(TNF-α),白细胞介素-6(IL-6),C-反应蛋白(CRP),髓磷脂碱性蛋白(MBP)水平。结果:与正常组比较,模型组p-JNK及Caspase3蛋白和JNK mRNA的表达明显升高,IGF-1 mRNA的表达明显降低,血清TNF-α,IL-6,CRP,MBP的水平明显升高(P<0.05);与模型组比较,通络糖泰方能一定程度下调p-JNK及Caspase3蛋白和JNK mRNA的表达,上调IGF-1 mRNA的表达,明显升高坐骨神经传导速度,明显降低血清TNF-α,IL-6,CRP,MBP的水平(P<0.05)。结论:通络糖泰方可能通过抑制高糖应激诱导的JNK信号转导通路的异常激活,改善细胞凋亡、胰岛素抵抗、炎症反应、下游靶基因的异常表达等病理过程,达到防治DPN发生发展的作用。

Objective： To investigate the effect of Tongluo Tangtai power（ TLTTP） on JNK signaling pathway in rats of diabetic peripheral neuropathy（ DPN）. Method： Totally 70 GK rats were selected to set up the rat model of spontaneous DPN induced by high fat diet. the 50 successfully modeled DPN rats were randomly divided into five groups,namely model group,TLTTP high-dose group,TLTTP group,TLTTP Low-dose group,and metformin ＋ methycobal group,with 10 rats in each group; another 10 male GK rats were selected in normal group. All of the rats were detected for the expressions of p-JNK and Caspase-3 protein,JNK and Insulin-like growth factor 1（ IGF-1） mRNA,sciatic nerve conduction velocity,and the levels of tumor necrosis factor-α（ TNF-α）,interleukins-6（ IL-6）,C-reactive protein（ CRP）,myelin basic protein（ MBP） in serum after 4weeks of intragastric administration. Result： Compared with the normal group,the model group showed significant increase in p-JNK and Caspase-3 proteins,JNK mRNA expression,and serum TNF-α,IL-6,CRP and MBP levels,and significant decrease in IGF-1 mRNA expression（ P〈0. 05）; TLTTP could down-regulate the expression of p-JNK, Caspase3, JNK mRNA,increase the expression of IGF-1 mRNA,significantly improve sciatic nerve conduction velocity,and significantly reduce the levels of TNF-α, IL-6, CRP, MBP in serum.Conclusion： TLTTP may prevent the development of DPN by inhibiting JNK signaling pathway,mitigating ell apoptosis,IR,inflammation reaction and abnormal expression of downstream target genes.