Experimental Study On The Effect Of JNK On RANKL/OPG On Osteoclast Differentiation

Objective: To inhibit the JNK signaling pathway by adding different concentra tions of c-Jun N-terminal kinase(JNK)specific inhibitor SP600125,and to explore the related regulation of mature osteoclast(OC)formation.Methods: SD rat denta l follicle cells(DFCs)were isolated and cultured.CCK-8 was used to detect the e ffects of JNK inhibitor SP600125 at concentrations of 0,5,10,15,and 20 ?mol/L on the proliferation of DFCs.Bone marrow macrophages(BMMs)of C57 mice w ere isolated and cultured,and DFCs and BMMs were co-cultured according to a c ertain ratio,and inhibitors at concentrations of 0,5,10,15,and 20 ?mol/L were s eparately treated.After 72 h,real-time quantitative PCR was used to detect the expr ession changes of nuclear factor-kappaB receptor activator ligand(RANKL)and ost eoprotegerin(OPG)genes in DFCs.After 7 days and 9 days,the anti-tartaric acid phosphatase(TRAP)staining was observed.The number of OCs was taken;after 9 days,the bone-grinding tablets were taken out,and the shape and number of bo ne sag formation were observed by electron microscopy.Results: CCK-8 results: J NK inhibitor SP600125 had no significant effect on the proliferation of DFCs in th e concentration range of 0-20?mol/L(P>0.05).The results of PCR showed that aft er 72 hours of treatment,the expression of RANKL gene in DFCs decreased,OPG increased,and the difference was statistically significant(P<0.05),but the expressi on of RANKL was the weakest at 15?mol/L,the most OPG.Strong expression.T he results of TRAP staining showed that OC was produced in each group,and the staining was obvious.The OC number increased significantly on 9d compared wit h 7d.Compared with the control group,the OC number decreased in each group with inhibitors,and the difference was statistically significant(P<0.05).The results of electron microscopy showed that bone resorption lacuna was observed in each experimental group,and collagen fibers were visible in the lacuna.The formation of bone lacuna in the control group was significantly higher than that in the treat ment group(P<0.05).The results showed that staining and qRT-PCR were perform ed.Consistent.Conclusion: In vitro DFCs,the specific inhibitor SP600125 inhibits JNK signaling pathway and changes the expression of RANKL and OPG,which has a certain negative regulation effect on the formation and activation of osteoclas ts,suggesting that JNK signaling can affect tooth eruption.Provide research basis f or osteoclast-related oral diseases such as bone remodeling.