Expression And Significance Of TMEM100 In Neuropathic Pain Model

Objective:Discogenic pain,one of the most common orthopedic pain,is low back pain caused by irritation of pain receptors following disc degeneration or annulus fibrosus rupture.There are many misconceptions about discogenic pain,and misdiagnosis and mistreatment are serious in the vast underdeveloped areas.It is difficult to get early,effective and standardized diagnosis and treatment.Although intervertebral disc degeneration is the main cause of discogenic pain,in clinical diagnosis,treatment and scientific research,intervertebral disc degeneration has always been the focus of research,ignoring that pain is the urgent problem that affects the quality of life of patients.If the pain early intervention,not easy to delay no more,into chronic pain,chronic pain stimulates the central nervous system pathological refactoring,make progress more and more difficult to control pain,to cause great mental and physical trauma patients,often accompanied by physiological,psychological and social function change,not only affects the body function,more seriously affect the quality of life.Therefore,it is of great significance to find the key mechanism of chronic discogenic pain regulation and early effective targeted intervention.This study aims to study the expression of TMEM100 and its corresponding significance,so as to lay the groundwork for targeted pain intervention.Methods:A total of 48 male SD rats were included in this study and randomly divided into 4 groups:Normal group,Sham group,CCI(Chronic constriction injury)group,and TNI(Tibial Nerve injury)group,with 12 rats in each group.Paw withdrawal mechanical threshold(PWMT)and Paw withdrawal thermal latency(PWTL)were measured continuously until14 days after surgery,and changes in general behavior and pain behavior of rats were observed and recorded.After observing the changes of pain behavior,the rats were sacrificed 14 days after the establishment of the rat model.The L4-6 DRG of the rats was taken,and the changes in the transcription level and protein expression level of TMEM100 in L4-6 DRG of each group were detected by qRT-PCR,Western Blot,immunohistochemistry and immunofluorescence staining.Results:1.Behavioral results: CCI group and TNI group showed significant behavioral changes,such as foot contraction,licking,foot suspension and foot valgus.2.The results of pain behavior: PWMT in CCI group began to decrease significantly on the 3rd day after surgery,and decreased to the lowest level on the 10 th day after surgery,and continued until the 14 th day after surgery;PWMT in TNI group began to decrease significantly on the first day after surgery,and decreased to the lowest level on the seventh day after surgery,which lasted until the 14 th day after surgery.PWMT and PWTL in TNI group and CCI group were lower than those in Normal group and Sham group,with statistical differences(P < 0.05).There were no significant differences in PWMT and PWTL between Normal group and Sham group(P > 0.05),while there were no significant differences in PWMT and PWTL between CCI group and TNI group(P >0.05).3.QRT-PCR results: m RNA expression in TNI group and CCI group was significantly decreased compared with Normal group and Sham group(P < 0.05).There was no difference between Normal group and Sham group(P>0.05).There was no difference between TNI group and CCI group(P>0.05).4.Western Blot results: Compared with Normal group and Sham group,the expression of TMEM100 protein in TNI group and CCI group was significantly decreased(P < 0.05).There was no difference between Normal group and Sham group(P > 0.05).There was no difference between TNI group and CCI group(P>0.05).5.Immunohistochemical results: Compared with Normal group and Sham group,the expression of TMEM100 in TNI group and CCI group was weakly positive and low(P <0.05).There was no significant difference between the other groups(P>0.05).6.Immunofluorescence staining results: compared with Normal group and Sham group,the fluorescence intensity in TNI group and CCI group was significantly decreased(P<0.05).There was no significant difference between the other groups(P>0.05).Conclusion:We successfully constructed CCI and TNI models,and the data showed that TMEM100 was down-regulated in DRG of the above models,which may be involved in the occurrence and transmission of pain.