Study On The Mechanism Of The Intracellular Interleukin-1α Regulation Of The Production Of IL-8Induced By Chlamydia Trachomatis

Chlamydia trachomatis (Ct) is an important sexually transmitted diseases pathogen, which can cause cervicitis, salpingitis and pelvic inflammatory disease, even can lead to abortion, ectopic pregnancy, infertility and other serious complications. Ct infection can stimulate epithelial cells to produce a large number of inflammatory cytokines, involved in regulating the host’s defense function. The inflammatory response caused by Ct infections was the main cause of disease and the IL-1α is an important inflammatory mediator which help the resistance to infection, it can induce the production of inflammatory markers such as IL-1α, IL-6and IL-8.Objects:Exploring the important role of nuclear signal of intracelluar IL-1alpha regulate IL-8production induced by chlamydia trachomatis. The other is how Hax-1interact with IL-1alpha involved in regulate inflammation induced by chlamydia trachomatis.Methods:1. ELISA and IFC tested the production and localization of inflammatory cytokines induced by Ct infection. siRNA technology was apply to inhibit the expression of endogenous IL-1alpha of HeLa229cells, establish an endogenous IL-1alpha expression HeLa229stable cell line. 2. Constructing pro-IL-la plasmid, transfect the stable cell line, ELISA to detect the effect of exogenous IL-1alpha on the produce of the inflammatory cytokines IL-8.3. Western blot and cell immune fluorescence detect the Hax-1protein expression and subcellular localization in cells after Ct infection in HeLa229. Immunofluorescence test the colocazation of Hax-1and IL-1alpha after Ct infection4. Appling siRNA technology to knock down the express of Hax-1, ELISA to detect the effect of silence of Hax-1on the production of IL-8.Results:1.44h after Ct infection HeLa229cells can produce inflammatory cytokine IL-1α and IL-8, the IL-1α is mainly expressed intracellular, IL-8is mainly expressed extracelluar.2. IFC dectection proved that IL-1α can move into the nuclear after Ct infection, with the production of IL-8.3. ELISA detection screening HeLa229stable cell lines S2C2,44h after Ct infection, compared with the normal HeLa229cells, expression of IL-8of stable S2C2cells were significantly reduced.4. ELISA dectection proved that S2C2cells can express pro-IL-1α, the transfection of pro-IL-1α induce the expression of IL-8.5. Western blot proved that48h after the Ct infection, Hax-1expression was significantly increasing.6. Immunofluorescence proved that44h after Ct stimulate, both Hax and IL-1alpha move into the nuclear.Appling siRNA technology down regulate the expression of Hax-1has no effect on IL-8production induced by Ct infection.Conclusion:1. Production of IL-8depends on intracelluar IL-1alpha on the early stage of Ct infection.2. The Ct infection can increase the expression of Hax-1, stimulate protein IL-1alpha and Hax-1move into the nucleus on the same time.3. Down regulate of Hax-1does not affect the production of IL-8induced by IL-1alpha.