人健康和炎性牙槽骨成骨细胞COX-2、PGE2、OPG,RANKL的表达

Expressions and Reciprocity of COX-2,PGE2,OPG and RANKL in Healthy and Inflamed Human Osteoblasts

目的:初步探讨COX-2、PGE2、OPG和RANKL在牙周炎发病机制中所起的作用及其相互关系。方法:采用组织块法对人健康和牙周炎性牙槽骨进行体外培养和传代,加入含100μg/LrhOPG的不含胎牛血清的DMEM2mL。严格按照ELISA试剂盒说明进行操作,获得各指标的检测数值。结果:牙周炎组RANKL、PGE2、COX-2的表达较健康组明显增高,OPG的表达较健康组明显降低。加入rhOPG后,牙周炎组RANKL、PGE2、COX-2的表达明显降低;OPG表达明显增加。健康组RANKL、PGE2、COX-2表达明显降低;OPG表达增加。结论:RANKL、PGE2、COX-2可促进牙周炎的发生、发展,而OPG对牙周炎的发生、发展可起抑制作用,同时表明人工重组OPG可能协同其内源性OPG来共同抑制RANKL、PGE2、COX-2的活性。

Objective：To investigate the expressions of the COX-2,PGE2,OPG and RANKL in healthy and inflamed humans osteoblasts,and to compare the levels of COX-2,PGE2,OPG and RANKL after adding the recombinant human osteoprotegerin（rhOPG）into the cell culture so as to clear whether the COX-2,PGE2,OPG and RANKL may play a role in periodontitis pathogenesis,and to provide the foundation of appling rhOPG in the treatment of periodontitis.Methods：The healthy and inflamed osteoblasts were grown from tissue explants and cells between the 5th and 6th passage in culture were used.Aliquots（2ml）of cells at a density of 1.5×106/ml were seeded into 12-well cell culture plates with 2ml media containing 10% FBS for 2days.After the media were removed,the wells were washed twice with PBS,2ml media（FBS-free）was replaced and incubated again for 24 hours.100ng/ml OPG media（FBS-free）were added into wells.The culture media were collected and stored similarly.According to the ELISA requirements,we carried out the operations strictly.Results：The expressions of RANKL,PGE2and COX-2in periodontitis groups were increased obviously than those in healthy group and the expression of OPG were reduced obviously in periodontitis group than those in healthy group.To add the rhOPG into the cell culture plates,the expressions of RANKL,PGE2,COX-2were reduced obviously and the expression of OPG were increased obviously in periodontitis groups.In healthy group,the expressions of RANKL and PGE2were reduced obviously.The expression of COX-2were reduced（P0.05）.The expression of OPG were increased.Conclusion：The results showed that the RANKL,OPG,PGE2and COX-2were related closely with pathogenesis of periodontitis.The action of rhOPG together with the synergistic action of the endogenous OPG may reduce the expressions of RANKL,PGE2and COX-2in osteoblasts of periodontitis.