Mechanism Exploration Of Hirsutella Sinensis Mycelium In Regulating Immunity And Ameliorating Pulmonary Fibrosis

Fibrosis is a natural process showing excessive production of fibrous connective tissue during an organ or tissue repair and react.Acute and chronic inflammation often cause fibrosis,resulting in epithelial cells damage and excessive release of inflammatory mediator including cytokines,chemokine and other factors.Besides,inflammation influence immune imbalance and interfere with tissue repair,tissue regeneration and fibrosis.Pulmonary fibrosis(PF)is a chronic,irreversible and debilitating lung disease leading to loss of pulmonary function with dyspnea and hypoxemia,and finally respiratory failure and death.Recently,the prevalence and mortality of pulmonary fibrosis are increasing,but owing to its complicated mechanism of pathology,there is no effective cure.Cordyceps sinensis,as a traditional medical fungi,is an entomogenous fungus whose mycelia grow inside larvae of Lepidoptera moth,forming a complex of the larva body and dry stroma.According to reports,Hirsutella sinensis mycelium(HSM)is an antioxidant and an inhibitor of angiogenesis,HSM has been used to cure respiratory diseases.Some reports show that HSM can ameliorate fibrosis in the liver,pulmonary and kidney through inhibition of TGF-β1 expression and promotion collagen degradation.Nevertheless,the role of HSM in resistance to pulmonary fibrosis and its mechanism are unclear.So our research evaluate the effect of HSM on ameliorating lung fibrosis and discuss the underlying molecular mechanism.1、HSM could ameliorate CLP-induced sepsis through an inhibition of inflammation and regulation immunitySepsis is a systemic inflammatory response to infection triggered by Gram-positive and/or-negative organisms,sepsis-related morbidity and mortality remain intractable problems in critically ill patients.Cecal ligation and puncture(CLP)is an experimental model which produces all the pathologic consequences of sepsis,so,we used the CLP-induced sepsis model to exam the effect of Hirsutella sinensis mycelium(HSM)on inflammatory and immunoregulation during sepsis’ development.The results revealed that HSM can effectively inhibit production of pro-inflammatory cytokines and pro-fibrosis cytokines,reduced the histopathologic alterations and influence the amount of immune cells,thus showed a good therapeutic effect on the model of sepsis,suggesting that HSM has immunomodulatory and anti-inflammatory capacity.Besides,CLP cause lung injury,thus induce pulmonary fibrosis,and HSM display therapeutic effect of on mice’s pulmonary fibrosis.All those suggest HSM may achieve the goal of improving lung fibrosis through inhibition of alveolitis during the earlier period.2.HSM could ameliorate bleomycin(BLM)-induced pulmonary fibrosisBLM-induced pulmonary fibrosis is a more ideal experimental model.To explore whether HSM has an anti-fibrosis activity,we used the BLM-induced pulmonary fibrosis model to analyze the anti-fibrosis activity of HSM during lung fibrosis’ development.Amazedly,HSM could markedly suppress the amount of hydroxyproline(HYP)in pulmonary section,inhibit pro-inflammatory cytokines’ and pro-fibrosis cytokines’ production and improve the injured organ.Besides,HSM reduce macrophages’ chemotaxis and adjust immune imbalance.It was displayed a well therapeutic effect of HSM on mice’s pulmonary model.3、HSM affect incidence of pulmonary fibrosis through inhibition phosphorylation of mTOR p70S6KIn order to investigate the inhibitory mechanism of HSM on pulmonary,we used the Q-PCR assay to analyze the inhibitory effect of HSM on TGF-pl-induced fibrosis associated molecules’ mRNA expression from A549.From the Q-PCR assay’s result,HSM inhibited the vimentin and fibronectin expression from TGF-β1-induced A549 cells.Besides,we used Western Blot assay to analyze the influence of HSM on the productions of phosphorylation of p70S6K from pulmonary sections in mice and TGF-β1-induced A549 cells,the result showed HSM effectively inhibit phosphorylation of p70S6K.All the results suggest that HSM could be a promising therapeutic agent for pulmonary fibrosis diseases,and the effect is attributed to the influence of mTOR pathway by down-regulating of p70S6K.In summary,we have found HSM can effectively regulate immunocytes,function,suppress inflammation and ameliorate pulmonary fibrosis.Meanwhile,the effect of HSM on formation of lung fibrosis may be associated with inhibition phosphorylation of mTOR p70S6K.