A Preliminary Study On The Effect Of TGF-β1 On Inflammatory Cytokines In Rat Bronchial Fibroblasts

Objective:Rat bronchial fibroblasts were obtained by culturing rat bronchial fibroblasts in vitro.The possible effects of TGF-β1 on IL-6,IL-8 and CCL20 secretion from rat bronchial fibroblasts were preliminarily explored.Methods:1.Isolate and culture normal rat bronchial fibroblasts in vitro.Basing on cell morphology,tissue origin,and identification of rat bronchial fibroblasts using immunofluorescence identification,select third-generation cells for experiments.2.Interfering rat bronchial fibroblasts with TGF-β1 concentrations of Ong/ml(control group),5ng/ml,10ng/ml,and 20ng/ml for 2h,12h,24h and 48h respectively,and collecting cell cultures from each group.The supernatant was tested for levels of IL-6,IL-8 and CCL20 in the supernatant of bronchial fibroblasts of each experimental sample by ELISA method.Results:1.Rat bronchial fibroblasts were successfully cultured in vitro.The cells were long spindle-shaped,triangular or polygonal cells.Basing on cell morphology,tissue origin,and immunofluorescence identification,the cells were indeed rat bronchial fibroblasts.After passage,the cells grew vigorously,were in good condition and stable character.It may be an ideal experimental model of bronchial fibroblasts.2.By ELISA method,under the condition of Ong/ml TGF-β1,the expression of IL-6 in rat bronchial fibroblasts at 2h,12h,24h,48h were 26.412±0.471ng/1,26.912±0.274ng/l,28.046±0.565ng/l,28.862±0.695ng/l,the expression levels of CCL20 were 10.163±0.261ng/l,10.396±0.429ng/l,11.197±0.381ng/l,11.771±0.168 ng/l,the expression levels of IL-8 were 772.633±1.989ng/l,775.826±2.306ng/l,781.454±2.589ng/l,789.492±1.840 ng/1.3.Compared with the Ong/ml control group,except for the TGF-β1 5ng/ml group,the content of IL-6 in the bronchial fibroblast supernatant of rats increased during the 2h period(P<0.05).The content of IL-6 in the bronchial fibroblast supernatant of rats at each time period was lower than that in the control group(P<0.05).4.Compared with the Ong/ml control group,except for the TGF-β1 5ng/ml group,the content of CCL20 in the bronchial fibroblast supernatant of rats was not significantly different within 2 hours(P>0.05).The content of CCL20 in the bronchial fibroblast supernatant of rats at each time period was lower than that in the control group(P<0.05).5.Compared with the Ong/ml control group,except for the TGF-β1 5ng/ml group,the content of IL-8 in the supernatant of rat bronchial fibroblasts increased within 24 hours(P<0.05).The content of IL-8 in the bronchial fibroblast supernatant of rats at each time period was lower than that in the control group(P<0.05).Conclusion:1.In this experiment,primary culture of rat bronchial fibroblasts was successfully obtained.2.Bronchial fibroblasts could be used as source cells for IL-6,IL-8 and CCL20.3.TGF-β1 could induce and inhibit the release of IL-6 and IL-8,but it is mainly inhibited in general,and has obvious time and concentration dependence.TGF-β1 inhibited the expression of rat bronchial fibroblast CCL20 in a concentration and time dependent manner.