The Effectiveness Of Cultured Cordyceps Sinensis On Pulmonary Fibrosis Induced By Bleomycin In Rats

Background and Objective:Idiopathic pulmonary fibrosis (IPF) is a chronic,diffuse interstitial pulmonary disease associated with a characteristic histologic appearance of usual interstitial pneumonia (UIP). The principal symptoms are described as dry cough and dyspnea. However, IPF generally progresses relentlessly and carries the poorest prognosis of the chronic IPF with a median survival of 3-5 years after diagnosis, there is no effective way to cure this disease yet. The current standard therapy recommended by ATS (American Thoracic society) for IPF involves treatment with corticosteroids and other immunosuppressive/cytotoxic agents. However, the beneficial effects of these agents are not well established, and adverse effects are often seen in the long treatment with glucocorticoid.Cordyceps sinensis is a medicinal fungus of Traditional Chinese Medicine. There are a wide range of reported uses of Cordycep sinensis in the literature. However, the production of Cordyceps sinensis is so limited that can not be widely used.Cultured Cordyceps sinensis and natural Cordyceps sinensis have similar chemical composition. Cultured Cordyceps sinensis possesses anti-inflammatory,anti-hypoxia,anti-tumor effect function and regulating the endocrine system, enhanced immune function, which has a protective effect on the kidney,lung,liver and other organs. Recently, cultured Cordyceps sinensis has some beneficial effects on pulmonary fibrosis. So our experiments are to study the treatment effects and mechanism of cultured Cordyceps sinensis on pulmonary fibrosis,and the treatment effects of cultured Cordyceps sinensis combined with glucocorticosteroid on experimental pulmonary fibrosis in rats induced by Bleomycin. The rats were respectively intratracheally instilled with Bleomycin to set up a pulmonary fibrosis model.PartⅠThe effectiveness of cultured Cordyceps sinensis on pulmonary fibrosis induced by Bleomycin in ratsAim: To study effectiveness of cultured Cordyceps sinensis on pulmonary fibrosis induced by Bleomycin in rats.Methods: Fifty rats were randomly divided into five groups, including control group (Ctr group), model group(Blm group), cultured Cordyceps sinensis group(Ccs group),prednisone group(Pred group).On experimental day 0,the rats were respectively intratracheally instilled with Bleomycin, and rats in the Ctr group and Blm group with the same volume of normal saline. One day after the injection, Ccs group and Pred group was respectively given to rats daily by gastric gavage, while the same volume of normal saline was given to those in the Ctr group and Blm group.During the experimental,the rats were observed the general conditions and measured the body weight.On 28th d, bronchoalveolar lavage fluid(BALF) and lung tissue were collected. Histological changes of the lungs were evaluated by HE stain , Masson's trichrome stain. Collagen content of the lung tissue was assessed by hydroxyprolin concentration.Result: (1)Compared to Blm group, the Ccs group improved the body weight loss and general conditions;(2) Compared to Blm group,pulmonary fibrosis and collagen deposition were alleviated in Ccs group and Pred group ;(3) Compared to Blm group,hydroxyprolin concentrations were decreased in Ccs group and Pred group.Conclusion:Ccs group can alleviate pulmonary fibrosis,improve the general conditions and the body weight loss induced by Bleomycin in rats.PartⅡThe mechanism of cultured Cordyceps sinensis on pulmonary fibrosis induced by Bleomycin in ratsAim: To study mechanism of cultured Cordyceps sinensis on experimental pulmonary fibrosis in rats induced by Bleomycin.Methods: Eighty rats were randomly divided into14d group and 28 d group. 14 d group was divided into four groups, including control group(Ctrl4d group),model group(Blml4d group), prednisone group(Predl4d group) , and cultured Cordyceps sinensis group( Ccsl4d group). 28d group was divided into four groups, including control group(Ctr28d group),model group(Blm28d group), prednisone group(Pred28d group) , and cultured Cordyceps sinensis group( Ccs28d group). On experimental day 0,the rats were respectively intratracheally instilled with Bleomycin, and rats in the control group and model group with the same volume of normal saline. One day after the injection, cultured Cordyceps sinensis was given to rats daily by gastric gavage, and prednisone was given to the prednisone group, while the same volume of normal saline was given to those in the control group and model group.At 14d, bronchoalveolar lavage fluid(BALF) and lung tissue of l4 day group rats were collected.The contents of TNF-αin BALF were measured. The left lung was subjected to HE staining, iNOS immunohistochemical test, and homogenized for biochemical assays of MDA, GSH contents. At 28d, bronchoalveolar lavage fluid(BALF) and lung tissue of 28 day group rats were collected. The contents of TGF-β1 protein in BALF were measured. The lung were subjected to CTGF immunohistochemical test. Result: (1)Compared to Blml4d group, alveolitis were alleviated in Ccsl4d group,and the iNOS protein expression in lung tissue decreased.The MDA,TNF-αcontents decreased,and the contents of GSH,SOD increased in Ccsl4d group .(2)Compared to Blm28d group, the TGF-β1 contents and the CTGF protein expression in lung tissue decreased in Ccs28d group.Conclusion:(1) Cultured Cordyceps sinensis alleviated alveolitis,which was through decreasing MDA,TNF-αcontents and iNOS protein expression, increasing the GSH,SOD contents.(2) Cultured Cordyceps sinensis alleviated pulmonary fibrosis,which was through decreasing the TGF-β1 contents and the CTGF protein expression.(3) The mechanism by which cultured Cordyceps sinensis resists pulmonary fibrosis is involved in alleviating alveolitis, enhancing the power of elimination oxygen radical,reducing ROS-mediated TGF-β1 activation and inhibiting the production CTGF to synthesis the extracellular matrix(ECM).PartⅢThe effectiveness of cultured Cordyceps sinensis combined with glucocorticosteroid on pulmonary fibrosis induced by Bleomycin in ratsAim: To study the treatment effects of cultured Cordyceps sinensis combined with glucocorticosteroid on experimental pulmonary fibrosis in rats induced by Bleomycin.Methods: One hundred and five rats were randomly divided into seven groups, including control group(Ctr group), model group(Blm group), cultured Cordyceps sinensis group(Ccs group),conventional-dose prednisone group(C-Pred group), low-dose prednisone group(L-Pred group),cultured Cordyceps sinensis combined with conventional dose prednisone group(C-Com group),cultured Cordyceps sinensis combined with low dose prednisone group(L-Com group). On experimental day 0,the rats were respectively intratracheally instilled with Bleomycin, and rats in the Ctr group and Blm group with the same volume of normal saline. One day after the injection, cultured Cordyceps sinensis and glucocorticosteroid were respectively given to rats daily by gastric gavage, while the same volume of normal saline was given to those in the Ctr group and Blm group. During the experimental,the rats were observed the general conditions and measured the body weight.On 28th d, bronchoalveolar lavage fluid(BALF) and lung tissue were collected. Histological changes of the lungs were evaluated by HE stain , Masson's trichrome stain. Collagen content of the lung tissue was assessed by hydroxyprolin concentration.Lung expression of TGF-β1 protein was assessed by immunohistochemistry.Lung expression of PDGF-BB protein was assessed by Western blotting.The level of bFGF protein was measured by enzyme-linked immunosorbent assay (ELISA).Result:(1)Compared to Blm group, the Ccs group,Pred group,C-Pred group L-Com group,C-Com group improved the body weight loss and general conditions;(2)Compared to Blm group, pulmonary fibrosis were alleviated in Ccs group and C-Pred group,and PDGF TGF-β1 protein expression, hydroxyproline concentrations and protein bFGF were decreased.(3)The combination effect of C-Com group or L-Com group was augmented compared with using Cordyceps sinensis or prednisone group alone.Conclusion:(1)The combination use of cultured Cordyceps sinensis and prednisone ,which may be through down-regulating the factors promoting extraceular matrix(ECM) synthesis TGF-B1,PDGF and bFGF,has synergia effects in anti - fibrous degeneration and improving the body weight loss ,general conditions.(2) Cultured Cordyceps sinensis combined with low dose prednisone had the same anti-fibrosis effect as Cultured Cordyceps sinensis combined with conventional dose prednisone.