The Mechanism Of Long Non-coding RNA Linc00968 On Chondrocyte Inflammation And Apoptosis Through Regulating PTBP1/p66shc

Objective Osteoarthritis（OA）is a highly prevalent degenerative joint disease that affects the health of more than 500 million people worldwide and has become one of the leading causes of chronic disability in the elderly.The currently known pathological mechanism of OA is that various factors lead to inflammation and apoptosis of chondrocytes,which in turn lead to articular cartilage destruction,subchondral bone remodeling and osteophyte formation.However,these understandings are not enough to block the progression of OA,and Surgical treatment prognosis also has a variety of possible complications.The aim of this study was to explore the mechanism of long non-coding RNA（lnc RNA）linc00968 on chondrocyte inflammation and apoptosis through regulating PTBP1/p66 shc.Methods（1）Taking normal chondrocytes as the research object,the expression level of linc00968 was up-regulated by linc00968 plasmid;（2）Taking IL-1β-induced chondrocytes as the research object,the expression level of linc00968 was silenced by linc00968 si RNA;（3）Taking normal chondrocytes as the research object,linc00968 was up-regulated by linc00968 plasmid,and the expression level of PTBP1 was silenced by PTBP1 si RNA at the same time;（4）Taking normal chondrocytes as the research object,linc00968 was up-regulated by linc00968 plasmid,and the expression level of p66 shc was silenced by p66 shc si RNA at the same time;Western blot technology was used to detect the changes in the expression levels of COL II,MMP-3,MMP-13,and COX-2 at the protein level after the above various treatments.Combined with the experimental results of RNA-pulldown combined with mass spectrometry analysis and co-immunoprecipitation,the data were analyzed to obtainin conclusion.Results（1）The expression of lnc RNA linc00968 was significantly increased in the articular cartilage of OA patients;（2）After over expression of linc00968 in human articular chondrocytes,chondrocyte inflammation and apoptosis were promoted;knocking down of linc00968 in human articular chondrocytes partially alleviated chondrocyte inflammation;（3）In human articular chondrocytes,the protein binding to linc00968 was detected by RNA-pulldown combined with mass spectrometry,and it was found that linc00968 had the highest binding rate to PTBP1;（4）Over expression of linc00968 in human articular chondrocytes and knockdown of PTBP1 at the same time resulted in partial remission of chondrocyte inflammation caused by overexpression of linc00968;（5）Using Blast analysis,it was found that there are multiple complementary sequences between linc00968 and p66 shc m RNA;（6）The co-immunoprecipitation experiments of interacting proteins confirmed that PTBP1 and p66 shc were combined in human articular chondrocytes;（7）Over expression of linc00968 in human articular chondrocytes and knockdown of p66 shc at the same time resulted in partial remission of chondrocyte inflammation caused by overexpression of linc00968.ConclusionsOver expression of linc00968 in human articular chondrocytes promotes inflammation and apoptosis,possibly by regulating PTBP1/p66 shc.