Effect Of Yiqihuoxuetonghuo Recipe On The Expression Of Inflammatory Cytokines And Nerve Growth Factor In Rats With Diabetic Peripheral Neuropathy

Objective:This topic is to observe the effect of Yiqi Huoxue Tongluo Recipe on the expression of inflammatory cytokines ? nerve growth factor and its receptor in diabetic peripheral neuropathy rats,and to explore the molecular mechanism of Yiqi Huoxue Tongluo Recipe on the treatment of diabetic peripheral neuropathy.Methods:1.The SPF 120 male SD rats were selected,fifteen rats were randomly selected as the normal group,the remaining rats using high fat and sugar method of feeding in combination with small dose of STZ induced diabetic rat model was established.2.After the success of the model,the rats were randomly divided into model group,Yiqi Huoxue Tongluo Recipe high dose group,Yiqi Huoxue Tongluo Recipe middle dose group,Yiqi Huoxue Tongluo Recipe low dose group and positive control group.the rats in the high,medium and low dose groups were given daily dosage of 9.0?4.5 and 2.5g/kg,the rats in the positive control group were treated with cobalt ammonium(0.225mg/kg),the rats in the normal and model groups were given the same volume of distilled water for 8 weeks.3.During the administration of the rats,the general state of the rats was observed and the changes of blood glucose and body weight were measured.After dosing 8 weeks,the temperature threshold and sciatic nerve conduction velocity were measured.After the materials,the changes of morphology and ultrastructure of sciatic nerve were observed by HE staining and transmission electron microscopy.The levels of TNF-a,IL-1,MMP-1,MCP-1 and NGF in serum were detected by ELISA.The expression of NGF m RNA in sciatic nerve was detected by RT-PCR.Western blotting was used to detect the expression of p75 NTR and Trk A Protein.Results:1.Compared with the normal group,the blood glucose of each model group was significantly higher than that of the normal group(p<0.01),there was no significant difference between the treatment group and the model group(p>0.05),the blood glucose of rats was decreased before intervention(p<0.05,p<0.01).After modeling,the body weight of each group was slightly increased compared with that before model,there was no significant difference between the two groups(p>0.05).After intervention,the body weight of the normal group was significantly increased,in the model group,the body weight decreased and the body weight increased after the intervention.2.Compared with the normal group,the sciatic nerve tissue morphology and ultrastructural changes were changed in the model group,and the threshold of the tailing was significantly increased(p<0.01),while the sciatic nerve conduction velocity was significantly decreased(p<0.01).Compared with the model group,the threshold of the tailing temperature of the treatment group and the methylcobalamin group was significantly lower than that of the model group(p<0.05,p<0.01),at the same time,the rate of sciatic nerve conduction was significantly increased(p<0.05,p<0.01).3.The levels of TNF-a,IL-1,MMP-1 and MCP-1 in the serum of the model group were significantly higher than those in the normal group(p<0.05,p<0.01).Compared with the model group,the treatment group could inhibit the expression of cytokines,the difference was statistically significant(p<0.05,p<0.01).4.The expression of NGF in the model group was significantly lower than that in the normal group(p<0.01).The expression of NGF in the serum of rats was significantly higher than that of the model group(p<0.05,p<0.01).The relative expression of sciatic nerve m RNA in the model group was significantly lower than that in the normal group(P<0.05).Compared with the model group,the relative expression of m RNA in the dose group and the mecobalamin group was significantly decreased(p<0.05,p<0.01).5.The expression level of p75 NTR in the dorsal root ganglion of the model group was significantly higher than that in the dorsal root ganglion group,and the expression level of Trk A was significantly decreased(p<0.01).Compared with the model group,the expression of p75 NTR in the dorsal root ganglion of the rats in the high,medium and low dose groups was significantly lower than that in the model group(p<0.05).The expression of Trk A in the dorsal root ganglion of the treatment group was increased,the difference was statistically significant(p<0.01).Conclusion:1.High-fat and high-sugar diet combined with low-dose STZ-induced method to establish diabetic rats model,sciatic nerve tissue morphology and ultrastructural changes,pendulum temperature threshold significantly increased,while the sciatic nerve conduction velocity was significantly reduced,DPN rat model was established successfully.2.The expression of TNF-a,IL-1,MMP-1 and MCP-1 protein in serum of DPN rats was significantly increased,the expression of NGF protein and the relative expression of NGF m RNA in sciatic nerve decreased.The level of p75 NTR was significantly increased,while the expression level of Trk A was significantly decreased,indicating that inflammatory cytokines and nerve growth factor involved in the development of DPN.3.Yiqi Huoxue Tongluo Recipe can significantly reduce the serum TNF-a,IL-1,MMP-1and MCP-1 protein expression,and can significantly increase the expression of NGF and Trk A,reduce the expression of p75 NTR.4.Yiqi Huoxue Tongluo Recipe has the effect of improving the peripheral nerve tissue injury in DPN rats,reducing the expression of inflammatory factors and promoting the secretion of nerve growth factor may play an important role in it.