Celeeoxib Decrease Ij-ip Stimulated Cox-2 And I1-8 Production In Human Periodontal Ligament Fibroblasts

Objective: (1) To observe the expression of Cyclooxygenase 2 (COX-2) in Human periodontalligament fibroblasts (HPLFs) by the interference of IL-1βwith different concentrations. (2) Toobserved the inhibition of COX-2 and IL-8 producing by specific concentration ofIL-1β-induced HPLFs with varied concentrations of celecoxib，which offer experimentalfoundation for the clinical use of celecoxib to treat periodontitis.Methods: (1) Culture the primary HPLFs in vitro and identify them. (2) Experiments werecarried out with cells from the fifth to eighth passages. It’s randomly divided into control andexperimental groups, the control group is HPLFs in serum-free medium, the experimentalgroup is the same but adds 0.1ng/ml、0.5ng/ml、1ng/ml、5ng/ml、10ng/ml IL-1βseparately，Using immunohistochemical methods and ELISA to detect the expression of COX-2 to find theoptimal stimulatory concentration of IL-1β.（3）Using the cells from the fifth to eighthpassages, it’s divided into control and experimental groups randomly, the experimental group is10ng/ml IL-1βin serum-free medium,by adding different concentration of celecoxib(0μM、10μM、25μM、50μM、100μM), used DMSO (dimethyl sulfoxide, DMSO) to assistdissolving celecoxib, the control group use serum-free medium adds the same concentration ofDMSO. Applied with ELISA method to detect the amount of COX-2 and IL-8. All the datawere analized using single factor variance statistical analysis.Results: (1) Successful cultured in vitro,the HPLFs showed a typical fibroblast-likemorphology (long spindle or star shape) under the inverted microscopeimmunohistochemicalstaining: the HPLFs were negative in anti-keratin stainingnegativ, but positive in anti-vimentinstaining, this indicates that the HPLFs are derived from mesoderm. (2) In IL-1βstimulatedHPLFs to express the COX-2 experiments, the results of immunohistochemistry showed thatthe COX-2 was not expressed in the control group , but induced by IL-1βit shows positive incytoplasm and nulear membrane of the HPLFs and its expression level increased while theIL-1βconcentration on the rise,which is the same as the results in ELISA（P<0.05）.(3) In thecelecoxib inhibition on IL-1βstimulated HPLFs to express the COX-2 experiments: celecoxibcan inhibit the expression of COX-2 by HPLFs differently. The inhibition gradually becamestronger as the celecoxib level increased. In the celecoxib inhibition on IL-1βstimulated HPLFsto restrain the IL-1βexperiments: With different concentrations of celecoxib in eachexperimental group,the expression of IL-8 have different degrees of reduction,in addition to 10u M group,dosing groups and the group without adding group comparison,P<0.05；less than25μM, the inhibitory effect on IL-8 is enhanced with the increasing concentration, and theinhibition became strongest in 25μM,compared with control group,P>0.05. When celecoxib level exceeds 25μM, its inhibitory effect on IL-8 drops in a minor range,but the comparisonamong group, the difference was not statistically significant,P>0.05.Conclusions: (1) Tissue culture of periodontal ligament fibroblasts is easy to operate, which canprovide experimental basis for periodontitis. (2) As the increase of IL-1βconcentration in acertain range, its stimulation of HPLFs provided more COX-2. (3)Application of specificconcentrations of celecoxib can effectively reduce the inflammatory factor COX-2 and IL-8produced by IL-1βintervened HPLFs.