Effect Of MiRNA/HMGB1 On The Function And Autophagy Of Lumbar Cartilage Endplate Cells In Rats

Objective Degenerative disc disease is the most common disease in spine surgery,and cartilage endplate,as an important component of the structure and nutrition of the disc,its degeneration is the main cause of disc degeneration.The occurrence and development of cartilage endplate degeneration is a complex biological process involving autophagy,proliferation,apoptosis,invasion and migration.Although mirco RNA(micro RNA)and HMGB1(high mobility group box-1 protein high mobility protein-b1)are involved in the process of disc degeneration,whether there is a clear correlation between them and the related mechanism is still unclear.Therefore,we explore the relationship between mi RNA and HMGB1 and the function and autophagy of cartilage endplate cells,so as to provide new ideas and theoretical basis for the treatment of cartilage endplate degeneration.Methods At first,ATDC5 mouse chondrocytes were cultured and subcultured by 1% fetal bovine serum.After cell subculture,ATDC5 cells were divided into two parts for transfection.The first part was divided into the following six groups: blank control group(mock group),mirna-142-3p low expression group(inhibitor group),mirna-142-3p low expression control group(inhibitor NC group)Group B,blank control + HMGB1 inhibitor group(mock +antihmgb1 group),low expression + HMGB1 inhibitor group(inhibitor + antihmgb1 group),low expression control + HMGB1 inhibitor group(inhibitor NC group)+ Antihmgb1 group);the second part is set up as the following six groups: blank control group(mock group),mirna-142-3p overexpression group(MICs group),mirna-142-3p overexpression control group(MICs NC group),blank control + HMGB1 activator group(mock + HMGB1 group);overexpression + HMGB1 activator group(MICs + HMGB1 group),overexpression control +HMGB1 activator group(MICs NC + HMGB1 group).After the two parts of cell transfection,CCK-8 method was used to detect cell proliferation,scratch method to detect cell migration,Transwell chamber experiment to detect cell invasion,FITC anexin-v / PI double staining and flow cytometry to detect cell apoptosis,RT-PCR and Western blot were used to detect protein and m RNA(mainly including HMGB1,beclin-1,p62,Bax,bcl-2).Results1.In the first part of the experiment,it was observed that when the expression ofmirna-142-3p was down regulated in degenerated ATDC5 cells,the expression of HMGB1,beclin-1,p62,Bax and Bcl-2,Bcl-2 and m RNA were significantly increased,while the expression of beclin-1 and m RNA were significantly increased At the same time,the ability of cell proliferation,migration and invasion decreased,the rate of apoptosis increased,but the cell cycle did not change significantly.When HMGB1 inhibitor was added,cell proliferation,migration and invasion were restored,and apoptosis rate was decreased.2.In the second part of the experiment,it was observed that when the expression of mirna-142-3p was up-regulated in degenerated ATDC5 cells,the ability of cell proliferation,migration and invasion was restored.When HMGB1 activator was added,the cell proliferation,migration and invasion decreased significantly.Conclusion1.Mirna-142-3p can inhibit cell proliferation,migration and invasion and promote cell apoptosis by targeting HMGB1 in chondroendplate cells.2.The content of mirna-142-3p in cartilage endplate cells is closely related to the level of autophagy3.Mirna-142-3p and HMGB1 affect the proliferation,migration,invasion and apoptosis of chondroendplate cells by regulating the autophagy level.