| Pulmonary Interstitial Fibrosis (PIF) is a kind of disease from various insults to the lung. The key feature of IPF is the excessive proliferation of fibroblasts and excessive collagen production and deposition, followed by gradual distortion of normal lung tissue architecture, which finally leads to the lung dysfunction. In recent years, the incidence of PIF is fast increasing for the air pollution. However, the lack of effective therapy and the sustainable developmental potential of PIF make the five-year survival rate of PIF patients under 50%. Thus it is a Hot pot to look for the effective medicine for PIF.The modern medicine study showed that the loss control of TGF-βexpression, the imbalance of Th1/Th2 and the abnormal activation of CD40/CD40L system were contributed to the PIF. These results demonstrated that was a multi-factors induced lung disease. Thus the combined therapy strategy that simultaneous targeting multi-targets became the hot targets for PIF therapy and draw many researchers' attentions.Traditional Chinese medicine (TCM) was an important mine and it had a 2,500 year's history for traditional Chinese medicine to adopt combined treatment utilizing 100,000 formulae. Although they demonstrated good effectiveness in clinical practice, the dimness of effective constituent and mechanism of many formulae restricted their spreading. Therefore, with morden technology and combinding the multi-suject methods, it was necessary and possible to elucidate the effective constituent and mechanism of TCM and thereafter spreading their application.FeiKangLing (FKL) was a formula based on traditional Chinese meicine theories and it showed good clinical efficacy. The total efficacy was 68% of 58 clinical trials. For elucidating the effective constituent and mechanism of FKL and spreading its application, morden biotechnology, proteomics and pharmcological technology were adopted to explore the mechanism and molecular targets of FKL under the assistance of two National Natural Science Foundation of China.In in vitro study, serum pharmacology was adopted. And the supernatant of macrophage and TGF-βwere used as inducer to observe the protective effects of FKL on PIF with MTT, flow cytometry and confocal. The results showed that FKL can evidently inhibit both the supernatant of macrophage and TGF-βinduced human embryo pulmonary fibroblast proliferation. The further studies displayed that FKL increased Th1, decreased Th2 and decreased the inflammatory factors, and thus relieve the PIF. In vivo, FKL also decreased the inflammatory factors from macrophage in bleomycin induced rat fibrosis model. The histomorphology study exhibited that FKL could decrease the inflammatory cells invasion, the fibroblast cells proliferation and matrix deposition, and thereby inhibit fibrosis. These results suggestted FKL was able to inhibit PIF.The further studies demonstrated that the main active constituent of FKL was polysaccharide. In this paper, we integrate affinity chromatography, mass spectrum, immunoprecipitation, SDS-PAGE, western blot, co-localization, and SPR, and combinding the polysaccharide-chip to explore the mechanism and targets of FKL. Results showed that the polysaccharides can bind to TGF-βand RIP1, through which polysaccharides inhibited the signals of TGF-βand CD40 system, decreased the inflammatory factors, balanced Th1/Th2, and finally inhibited PIF.It is the first time that the mechanism of FKL were elucidated from in vivo, cellular and molecular level through compositive technology system. This study opens up a good source for the PIF treatment and possessed important theoretical value. |
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