The Effect Of Estrogen On Expression Of HSP27in HPLFs Induced By Lipoplysaccharide

Objective:(1)To construct the human periodontal ligament fibroblasts (HPLFs)model in vitro by primary tissue culture method.(2)The expression of heat shock protein27(HSP27) will be observed in HPLFs after the intervention of LPS.(3)The effect ofestrogen on expression of HSP27will be observed in HPLFs induced by LPS,andprovide the foundation of cytokine involves in periodontal desease.Methods:(1)To isolated and culture human periodontal ligament fibroblasts byprimary tissue culture method,immunohistochemistry method was used to assess thecells’ biology features.(2)The cells from the fifth to eighth passage was stimulatedwith0μg/ml、0.1μg/ml、1μg/ml、10μg/ml、50μg/ml、100μg/ml of LPSrespectively,and detected the expression of HSP27with immunehistochemicalmethods after24hours. The cells was stimulated with LPS which group shows thestrongest positive reaction,and ascertain the specifically concentration.(3)To choosethe fifth to eighth passage once more,and stimulated with the specificallyconcentration of LPS with0mol/L、10-9mol/L、10-8mol/L、10-7mol/L、10-6mol/Lof estrogen on cells respectively.To detecte the effect of estrogen on expression ofHSP27with immunohistochemical methods after24hours.(4)To collecte and analyzethe images and data by computer image analysis system.Both of the results wereanalyzed with single-factor variance analysis.Results:(1) To construct the human periodontal ligament fibroblasts model in vitrosuccessfully.The cells showed fusiform or stellate cells were observed under aninverted microscope. Immunohistochemical identification showed that the expressionof anti-keratin is negative, and anti-vimentin is positive,the cells showed biologicalcharacteristics of the fibroblast-like cells; After five generations of cell growth isstrong,slow cell proliferation after8generations.(2) Having LPS intervention HPLFs,immunohistochemistry to detect the expression of HSP27in the cells shows that:negative or weakly positive expression of HSP27in the normal HPLFs,but After LPSintervention,positive expression of HSP27in the cytoplasm of the cell,and withincreasing concentration of LPS intervention, the expression of HSP27in showing thetrend of increment.(3) Using immunohistochemical methods to detect17β-estradiolon the expression of proinflammatory HPLFs of HSP27: With the17β-estradiol concentration increased, the expression of proinflammatory HPLFs of HSP27in adecreasing trend.Conclusions:(1)The primary tissue culture method has simple operation and highsuccess rate,the human periodontal ligament fibroblasts model was the foundation ofexperiments.(2) After LPS-induced HPLFs of HSP27have positive expression withthe expression intensity of LPS-induced have dose-dependent relationship showedthat:HSP27involved in the development of periodontal tissue inflammation.(3)Theexpression of HSP27in the inflammation of HPLFs with the intervention of estrogenput forward that estrogen involvs in the anti-inflammatory of desease and could be thefoundation of diagnosis and treatment of periodontal desease.