| Objective Neuropathic pain is a hot and difficult research topic in recent years.At present,there is no effective way to prevent and cure neuropathic pain,which greatly affects the quality of life of patients.Postherpetic neuralgia is a typical neuropathic pain.Because of its high diagnostic accuracy and typical clinical manifestations,it is often regarded as a classic research disease of neuropathic pain.At present,the recognized pathogenesis of neuropathic pain mainly includes peripheral sensitization and central sensitization,of which central sensitization is the key point.Effective prevention or reversal of central sensitization is an important event in the treatment of neuropathic pain.Early diagnosis and timely intervene can greatly improve the quality of life of patients.Because the dorsal horn of the spinal cord is an important connect between the peripheral nerve and the central nerve,it is reasonable and necessary to study neuropathic pain at the level of the spinal cord,especially the dorsal horn of the spinal cord.Based on the proteomic detection in the serum of patients with postherpetic neuralgia,this study screened out the differential proteins in the serum of patients with non-neuropathic pain and patients with neuropathic pain,and further explored the mechanism at the level of animal spinal cord,with the purpose of revealing its role in the generation of neuropathic pain,looking forward to further clinical transformation,providing early and objective diagnosis and screening means for neuropathic painMethods Part I: Characteristics of differential protein expression in peripheral serum of patients with postherpetic neuralgia and comprehensive bioinformatics analysis Select patients without pain after acute herpes zoster and with postherpetic neuralgia,and divide them into control group and case group(PHN group),with 10 patients in each group.Take peripheral blood samples of patients.The differential proteins were screened by protein quantitative DIA technology,including the difference analysis of non-null values in the control group and PHN group,and the analysis of the existence of null values in the control group or PHN group.Further differential protein results were analyzed by bioinformatics.Part II: The expression of SH3BGRL3 in the spinal cord of neuropathic pain rats and its effect on mechanical pain sensitivity1.SD rats were divided into control group,sham operation group and CCI group.The chronic compression injury model of sciatic nerve was established for CCI group rats,and the mechanical threshold of rats in each group was observed and recorded,and the morphology of neurons in the dorsal horn of the spinal cord was observed and compared.2.The rats were divided into sham operation group,CCI-D3 group,CCI-D7 group,CCI-D14 group and CCI-D21 group.The rats in each CCI group were treated with CCI respectively,and the lumbosacral enlargement of the spinal cord was taken at the corresponding time(the 3rd,7th,14 th and 21 st days after operation)for Western Blotting detection;Western blotting was performed on the 14 th day in the sham group to observe the expression of SH3BGRL3 and its correlation with mechanical hyperalgesia.3.Take the spinal cord tissue of rats 14 days after CCI operation for immunofluorescence detection.4.The rats were divided into sham operation group,CCI group,CCI+si RNA-NC group and CCI+si RNA group.The sham operation was performed on the rats in the sham group,and the rats in the other groups were performed CCI operation.The intrathecal injection was given to CCI+si RNA-NC group and CCI+si RNA group.The CCI+si RNA-NC group was given negative control solution,and the CCI+si RNA group was given SH3BGRL3 specific si RNA.The PWT at each time point of the rats was recorded.The samples were taken for Western Blotting test on the 14 th day after CCI operation,to observe the effect of reducing the expression of SH3BGRL3 in the spinal cord on mechanical sensitivity in CCI rats. Part III: SH3BGRL3 participates in the downstream signal pathway of neuropathic pain regulation1.SD rats were randomly divided into sham operation group,CCI-D3 group,CCI-D7 group,CCI-D14 group(and CCI-D21 group).Rats in CCI groups were treated with CCI,and samples were taken at the corresponding time for Western Blotting detection;rats in sham group were treated with sham operation,and samples were taken on the14 th day for Western Blotting detection,and the expression of p-EGFR in the spinal cord of CCI rats and its correlation with mechanical sensitivity were observed.2.The rats were randomly divided into sham operation group,CCI group,CCI+si RNA-NC group and CCI+si RNA group.Sham operation was performed on rats in sham group,and CCI operation was performed on rats in other groups.Intrathecal injection of drugs was given to CCI+si RNA-NC group and CCI+si RNA group.Negative control solution was given to CCI+si RNA-NC group,and SH3BGRL3 specific si RNA was given to CCI+si RNA group.Samples were taken on the 14 th day after CCI operation to observe the effect of reducing the expression of SH3BGRL3 in the spinal cord on p-EGFR and downstream pathway protein.3.The rats were randomly divided into sham group,CCI+si RNA-NC group,CCI+si RNA group and CCI+si RNA+Colivelin group(STAT3 agonist).Sham group received sham operation.CCI+si RNA-NC group,CCI+si RNA group and CCI+si RNA+Colivelin group received CCI operation.CCI+si RNA-NC group,CCI+si RNA group and CCI+si RNA+Colivelin group received intrathecal injection intermittently.CCI+si RNA-NC group received si RNA control solution,and CCI+si RNA group received si RNA solution,CCI+si RNA+Colivelin group was given si RNA solution and Colivelin(the time interval between the two reagents was more than 4 hours).On the 14 th day,the spinal cord tissue at the lumbosacral enlargement of rats was taken to observe the effect of STAT3 agonist on the mechanical sensitivity caused by the down-regulation of SH3BGRL3.Results Part I 1.Among the selected subjects,there was no statistical difference between the PHN group and the control group in terms of age,sex,location,and course of disease.There was a significant difference in VAS scores between the PHN group and the control group.2.Carry out proteomic difference analysis on the serum samples of the two groups of patients.There are 25 proteins with statistical differences and 29 proteins with or without analysis.Part II1.Chronic compressive injury of sciatic nerve induced mechanical hyperalgesia in rats,that is,CCI can induce neuropathic pain in rats.2.Chronic compression injury of sciatic nerve leads to neuron injury of spinal dorsal horn in rats.3.The expression of SH3BGRL3 in the spinal cord of CCI rats increased and reached its peak on the 14 th day after CCI.4.SH3BGRL3 is co-located with neurons in the dorsal horn of the spinal cord,mainly expressed in the cytoplasm of neurons,and slightly expressed in the nucleus.5.Intrathecal injection of specific si RNA can reduce the expression of SH3BGRL3 in the spinal cord and relieve the mechanical sensitivity of CCI rats.Part III1.The expression of p EGFR in the spinal cord of rats after CCI showed an increasing trend and reached its peak on the 14 th day,which was significantly different from that of the sham operation group.2.Inhibition of SH3BGRL3 in rat spinal cord can reduce the expression of p EGFR.3.Knocking down SH3BGRL3 in the spinal cord of rats can down-regulate the expression of p STAT3.4.STAT3 agonist can reverse the reduction of mechanical sensitivity caused by down-regulation of SH3BGRL3.Conclusions:There is differential protein expression in peripheral blood of patients without pain after acute herpes zoster and with postherpetic neuralgia and differential protein participate in biological processes,molecular functions and cell components.The expression of SH3BGRL3 in the spinal cord of CCI rats increased,which was positively correlated with the neuropathic pain produced by CCI;SH3BGRL3 is co-localized in the neurons of spinal cord and mainly expressed in the cytoplasm;SH3BGRL3 is involved in the formation of neuropathic pain in rats.The expression of p EGFR in the spinal cord of CCI rats increased,and SH3BGRL3 participated in the occurrence of neuropathic pain through STAT3-related pathway. |
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