Research Of Differential Expression Of TMEM100 In Genetic Intervention

Objective:It is believed that neuropathic pain(NP)is caused by abnormalities of the nervous system itself.Primary nerve injury or abnormal function can lead to the occurrence of NP.In the diseases related to spinal degeneration,although the compression is relieved by surgery and other related methods.However,there are still a considerable number of patients,especially those who have not been timely,standardized and effective treatment,will still have residual pain,which may be related to the long-term compression of intervertebral disc and the neuropathic pain of central divine system remodeling,but not closely related to discogenic pain.Long-term nerve pain will have a great negative effect on the life and work of the patients with the disease,and even endanger the mental health of the patients.Therefore,it is very urgent and necessary to choose a feasible way to alleviate the harm caused by neuropathological pain to patients and improve their quality of life.Although the treatment of discogenic pain is more mature,there is no clear and effective plan for the treatment of neuropathic pain.Therefore,how to relieve the rational pain of neuropathy,To alleviate the symptoms of patients and improve the prognosis is the main research direction at the present stage.,we found that there was a significant difference in the expression of TMEM100(Transmembrane 100)between the neuropathic pain model and the normal group.The purpose of this experiment is to verify the difference of TMEM100 expression in different neuropathic pain models,and to study whether TMEM100-related genes can interfere with neuropathic pain and provide help for the future neuropathological pain mechanism and even accurate targeted therapy.Methods:The results of previous experiments showed that there were significant differences in general behavior,pain behavior,Western Blot,qRT-PCR,immunohistochemistry and immunofluorescence between CCI,TNI group and Normal,Sham group.There was no significant difference among the other groups(P > 0.05).After the establishment of the above model,the mechanical foot withdrawal threshold(PWMT)andthermal foot withdrawal latency(PWTL)and other general behavior and pain behavior were observed and recorded,and the statistical differences were analyzed.The rats were killed 28 days after operation,and bilateral L4-6 dorsal root ganglia(DRG)were obtained immediately.The expression of TMEM100 protein and its gene in each group was observed by qRT-PCR,Western Blot,immunohistochemistry and immunofluorescence,and the statistical difference was analyzed.Results:The results of previous experiments showed that there were significant differences in general behavior,pain behavior,Western Blot,qRT-PCR,immunohistochemistry and immunofluorescence between CCI group and Sham group and Normal group and Sham group.There was no significant difference among the other groups(P > 0.05).1.General behavioral results: in the CCI group and TNI group,the pain,licking,valgus and suspension of the foot in the virus injection group were better than those in the non-virus injection group.2.Pain behavior results:(1)PWMT results: the PWMT of T-CCI group and T-TNI group reached the lowest on the 5th day and 10 th day respectively,and was significantly higher than that of CCI and TNI group on the 21 st day.On the 28 th day,the performance was similar to that of Normal group and Sham group.(2)PWTL results:the PWTL of T-CCI group and T-TNI group reached the lowest on the 5th day and 10 th day respectively,and was significantly higher than that of CCI and TNI group on the 21 st day.On the 28 th day,the performance was similar to that of Normal group and Sham group.3.Western Blot results:T-CCI group and T-TNI group ‘s expression was higher than CCI group and TNI group,and there was statistical difference.The histone expression of Normal was similar to that of Sham(P > 0.05).The other group has no difference among(P > 0.05).4.qRT-PCR results:although the T-CCI group and T-TNI group’s expression was still lower than Normal group,but was prominently higher than the CCI group and TNI group’s expression(P < 0.05).There was no difference in expression between T-CCI and T-TNI group.The other group has no difference among(P > 0.05).5.Immunohistochemical results:the expression rate of protein in T-CCI group and T-TNI group was similar to that in Normal group,but significantly different from that in CCI group and TNI group.The expression rate of histone in Normal was the same as that in Sham.The other group has no difference among(P > 0.05).6.Immunofluorescence results:the average fluorescence intensity of T-CCI group and T-TNI group was significantly different from that of CCI group and TNI group,but no difference between Normal and Sham group.The other group has no difference among(P > 0.05).Conclusion:In this study,we verified the conclusions of previous experiments,that is,there are statistical differences between CCI model and TNI model in general behavior and pain behavior,as well as TMEM100 expression compared with Normal group and Sham group.After injection of TMEM100 gene virus group,by means of general behavior,pain behavior,Western Blot,qRT-PCR,immunofluorescence and immunohistochemistry,the expression of TMEM100 was significantly higher than that of non-virus injection group,and significantly lower than that of Normal group and Sham group.This proves that TMEM100 is involved in the process of neuropathic pain.The results showed that the expression of TMEM100 in neuropathic pain model was different from that in normal rats,and injection of TMEM100-carrying virus could improve neuropathic pain.However,the mechanism of the effect of TMEM100 on neuropathic pain is not clear,and it is not clear that TMEM100 is involved in the generation or transmission of neuropathic pain,which needs further study.