| Background Nonspecific low back pain(NLBP)refers to acute and chronic pain in the lumbosacral region excluding specific pathogenic factors,usually ranging from the lower edge of the ribs to the upper edge of the transverse striations of the buttocks,with or without thigh pain.The high incidence of NLBP and the huge consumption of medical resources have become a heavy social burden.At present,the commonly used treatment methods mainly include bed rest,drug therapy,physical / rehabilitation therapy,cognitive behavioral therapy,traditional Chinese medicine treatment and so on.Each therapy has its own advantages and disadvantages.Scraping is one of the commonly used characteristic therapies of traditional Chinese medicine.Because its operation is easy to learn,and treatment tools are simple and lightweight,it is easy to promote at the grass-roots level,and can effectively save medical resources.Scraping therapy can improve the microenvironment of the body by increasing local tissue temperature,increasing microcirculation and promoting energy metabolism,and play a role in dredging channels and collaterals,dispelling pathogens and detoxification.It has been reported that scraping has a significant effect on NLBP.At present,the research on scraping is mainly to observe the clinical efficacy,and most of the treatment measures are two or more.The related laboratory basic research is very few,and the potential mechanism remains to be clarified.By studying the therapeutic effect of scraping on rats with multifidus injury,and with the help of transcriptional sequencing technology,our research group intends to explore the potential mechanism of its therapeutic effect,so as to provide experimental data support for clinical promotion of scraping treatment of NLBP,and lay a foundation for further elucidating the mechanism of scraping therapy in the future.Objectives 1.To observe the effect of scraping intervention on the local tissue of healthy rats and evaluate its safety.2.To observe the effect of scraping on the local tissue of rats with lumbar multifidus injury and evaluate its curative effect.3.To explore the potential mechanism of the therapeutic effect of scraping.Methods 1.30 male SD rats aged 7 to 8 weeks were randomly divided into 5 groups:blank group,6h scraping control group,1d scraping control group,2d scraping control group and 3d scraping control group,with 6 rats in each group.Rats in the scraping control group were given scraping intervention after anesthesia,while rats in the blank group were grabbed and anesthetized simultaneously.The changes of skin surface temperature,mechanical pain threshold of hindfoot,skin and multifidus tissue were observed in each group.2.18 male SD rats aged 7 to 8 weeks were randomly divided into 3 groups: blank group,model control group and model group,with 6 rats in each group.The rats in the model group were locally injected with 0.5% bupivacaine solution,while the rats in the model control group were locally injected with normal saline,and the rats in the blank group were grabbed and anesthetized synchronously.The changes of mechanical pain threshold and multifidus tissue of hind foot were observed before and after modeling.3.54 male SD rats aged 7 to 8 weeks were randomly divided into 9groups: blank group,6h model group,1d model group,2d model group,3d model group,6h scraping group,1d scraping group,2d scraping group and 3d scraping group,with 6rats in each group.The rats in the scraping group were treated with scraping intervention24 hours after the establishment of the model,and the rats in the other groups were grabbed and anesthetized synchronously.The changes of skin surface temperature,hindfoot mechanical pain threshold,skin and multifidus tissue were observed in each group.4.Transcriptional sequencing was used to compare the difference of gene expression between scraping group and model group after 6 hours and 2 days of scraping.GO and KEGG enrichment analysis were used to screen the related differential signal pathways.q PCR and WB were used to verify some of the sequencing results to explore the regulatory effect of scraping on GLUT4/ glycolysis pathway and AMPK/mTOR/4EBP1 signal pathway.Results 1.After scraping intervention,the skin surface of healthy rats was continuous and intact,the skin ecchymosis basically disappeared on the 3rd day,the size and space of muscle cells basically returned to normal,the mechanical pain threshold of hindfoot decreased on the 2nd day,and there was no significant change in the rest of the time.2.After local injection of 0.5% bupivacaine solution for 24 hours,the injury of multifidus cells was obvious,the average cross-sectional area of multifidus decreased,the mechanical pain threshold of hindfoot decreased,and the intercellular space of multifidus increased slightly in the model control group,and there were no other pathological changes.3.Within 3 days after scraping treatment,the muscle cell and muscle bundle structure of the scraping group tended to recover compared with the model group,and the mechanical pain threshold of the hindfoot on the 2nd day after scraping treatment was higher than that of the model group.The skin surface temperature increased immediately after scraping and 6 hours after scraping.4.The results of transcriptional sequencing showed that 6 hours after scraping,there were 391 differentially expressed genes between the scraping group and the model group,including 322 up-regulated genes and 69 down-regulated genes,and 2 up-regulated genes and 1 down-regulated genes 2 days after single scraping.KEGG enrichment pathway analysis showed that 8 signal pathways,such as glycolysis/gluconeogenesis,starch and sucrose metabolism,carbon metabolism,and fructose and mannose metabolism,were concentrated in 6 hours after scraping,and ketone synthesis and degradation,methyl butyrate metabolism and glycosphingolipid biosynthesis were mainly concentrated in 2days after scraping.5.q PCR results: compared with the model group,the expression of Hk2,Pfkm,Slc2a4 and Bdh1 increased and the expression of Prkaa1 decreased 6 hours after scraping in the scraping group;the expression of Pfkm decreased and the expression of Bdh1 increased 1 day after scraping,but there was no significant difference in the expression of Hk2,Slc2a4 and Prkaa1;2 days after scraping,the expression of Hk2,Slc2a4 and Bdh1 increased,and the expression of Pfkm and Prkaa1 had no significant difference;3 days after scraping,the expression of Hk2 and Bdh1 decreased,and the expression of Pfkm,Slc2a4 and Prkaa1 had no significant difference.6.WB results: 6hours after scraping,the expression of glycolysis pathway related proteins: compared with the model group,the expression of GLUT4,HK2,PFKM,PKM and LDHA increased in the scraping group.The expression of AMPK pathway related proteins:compared with the model group,the expression of AMPKα and p-AMPKα decreased,the expression of p-mTOR and p-4EBP1 increased,and the expression of mTOR and 4EBP1 did not change significantly in the scraping group.Conclusions 1.The safety of scraping is proved,and the pathological changes of body tissue can be recovered in a short time.2.Scrapping treatment can effectively promote the repair process of injured multifidus.3.The results of transcriptional sequencing showed that scraping may play a therapeutic role through signal pathways such as glycolysis/gluconeogenesis,starch and sucrose metabolism,carbon metabolism,and fructose and mannose metabolism.4.The partial results of transcriptional sequencing verified by qPCR and WB showed that 6 hours after scraping,GLUT4/glycolysis pathway was activated,the gene and protein expression of related rate-limiting enzymes increased,AMPK/mTOR/4EBP1 signal pathway was regulated,AMPKα activity decreased,and mTOR and 4EBP1 activity increased. |
PDF Full Text Request