Protective Funtion Of Yiqi Huoxue Tongluo Recipe On Peripheral Nerve Of Db/db Mice And Its Effect On JNK Signaling Pathway

Objective:To observe the protective effect of Yiqi Huoxue Tongluo Recipe on db/db mice peripheral nerve and its correlation with JNK signaling pathway,and explore the protective mechanism of Yiqi Huoxue Tongluo Recipe on diabetic peripheral nerve,so as to lay a theoretical foundation for its rational clinical application.Method:40 db/db mice were randomly divided into 5 groups: model group,lipoic acid group,Yiqi Huoxue Tongluo Recipe high,middle and low dose groups,and 8 db/m mice served as normal group.Normal group and model group were given normal saline at10ml/kg,Yiqi Huoxue Tongluo Recipe groups were treated with Yiqi Huoxue Tongluo Recipe suspensions by 6.4g/kg,3.2g/kg and 1.6g/kg respectively and lipoic acid group was administered with lipoic acid suspension by 78mg/kg,once a day for 8 weeks.The general condition of mice was observed and recorded.At the end of eight weeks,the plantar thermal response time and sciatic nerve conduction velocity were measured;the ultrastructural changes of sciatic nerve in mice were observed by transmission electron microscope;ELISA(enzyme-linked immunosorbent assay)method was used to detect the content of inflammatory factor TNF-alpha and IL-1 beta in db/db mice serum,Western blot method was used to detect the proteins related to JNK signaling pathway in db/db mice dorsal root ganglion(DRG),and apoptosis genes transcript levels of P53 and Bax in mice sciatic nerve were detected by RT-PCR.Result:1.Effects of Yiqi Huoxue Tongluo Recipe on the structure and function of sciatic nerve in db/db mice Compared with the normal group,motor nerve conduction velocity was slower,plantar thermal reaction time was longer(P < 0.01),and the pathological injury of the sciatic nerve was serious in the model group mice;compared with the model group,Yiqi Huoxue Tongluo Recipe could improve the conduction velocity of sciatic nerve and shortened the plantar thermal reaction time(P < 0.01 or P < 0.05),significantly alleviated the pathological injury of sciatic nerve.2.Effects of Yiqi Huoxue Tongluo Recipe on inflammatory factors in db/db mice serum Compared with the normal group,the levels of IL-1? and TNF-? in serum of model group were increased significantly(P < 0.01);compared with the model group,Yiqi Huoxue Tongluo Recipe could decreased the levels of IL-1 beta and TNF-alpha in serum(P < 0.01 or P < 0.05),and reduced inflammatory reaction.3.Effects of Yiqi Huoxue Tongluo Recipe on JNK signaling pathway related proteins expressions in db/db mice Compared with the normal group,content of JNK pathway associated proteins(MEKK1,MLK3,MEK7,JNK,p-JNK)in dorsal root ganglion of model group mice increased(P < 0.01 or P < 0.05);compared with the model group,Yiqi Huoxue Tongluo Recipe could inhibit the expression of the JNK signaling pathway associated proteins(MEKK1,MLK3,MEK7,JNK,p-JNK)(P < 0.01 or P < 0.05).4.Effects of Yiqi Huoxue Tongluo Recipe on the transcript levels of pro-apoptotic genes P53 and Bax in the sciatic nerve of db/db mice Compared with the normal group,the transcript levels of downstream pro-apoptotic genes P53 mRNA and Bax mRNA of JNK signaling pathway in sciatic nerve of model group mice increased(P < 0.01);compared with the model group,Yiqi Huoxue Tongluo Recipe could depress the transcript levels of apoptosis genes P53 and Bax in sciatic nerve(P < 0.01 or P < 0.05).Conclusion:Yiqi Huoxue Tongluo Recipe could improve the structure and function of db/db mouse sciatic nerve,showed a protective effect on db/db mice peripheral nerve.The mechanisms may be related to the inhibition of abnormal activation of JNK signaling pathway,the inhibition of inflammatory response in db/db mice and the reduction of neuronal apoptosis.