Experimental Study On The Interventional Effect Of Shengxian Tongluo Decoction On Pulmonary Fibrosis

Background:IPF is a kind of serious disease with high disability and death rates.In recent years,the incidence of this disease is going to increase.The pathogenesis of IPF is not fully understood as yet,as a result it is hard in drug R&D.Most studies show that the injure of human alveolar epithelial cells,the form of fibroblasts and extracellular matrix increased deposition are the main pathological features.Finally,alveolar structure destroyed and formed totally pulmonary fibrosis and vesicular structures,which leaded to pulmonary function sustained,progressively damage.Besides long-term oxygen therapy,there are no effective medicines to treat IPF.Chinese medicine for treatments of IPF have unique advantage which can relieve clinical symptoms and stay lung function decline.In recent years.Through our clinical observations,our research group discovered that sinking of pectoral QI is the initial factor of this disease,from which exogenous pathogenic obstruction,chronic diseases into collateral combined with endogenous damp-heat,phlegm and blood stasis accumulateion make further obstruction in lung collaterals,as a result,sinking of pectoral QI aggravated pulmonary vascular obstruction increasingly,phlegm and blood stasis obstructed in lung collaterals seriously.The treatments of Shengxian Tongluo Decoction which can invigorate qi and ascend qi collapse,ventilate lung collaterals,resolve phlegm and promote blood circulation have achieved good clinical healing efficacy.Purpose:Observe the effect of Shengxian Tongluo decoction on the mTOR/p70S6K signal pathway and the MMPs/TIMPs system in lung tissue of rats with pulmonary fibrosis,and the impact on Col Type Ⅰ,Col Type Ⅲ,Col Type Ⅳ,HA,FN and other ECM protein levels.To investigate the relationship between mTOR/p70S6K signaling pathway and ECM protein deposition,the relationship between MMPs/TIMPs system and ECM protein deposition,and observe the effect of Shengxian Tongluo decoction on pulmonary function in rats.Methods:Eighty healthy male SD rats were randomly divided into blank control group(n=10),model group(n=12),Shengxian Tongluo decoction 3rd day intervention group(n=14),Shengxian Tongluo decoction 14th day intervention group(n=12),Shengxian Tongluo decoction 28th day intervention group(n=12),N-acetylcysteine 3rd day intervention group(n=14),N-acetylcysteine 14th-day intervention group(n=12),N-acetylcysteine 28th intervention Group(n=12).After modeling the corresponding time in the node(day 3,14,28)corresponding to the start interventions,intervention consecutive 90 days.After the end of the intervention period,the lung functions(FVC,FEV0.3/FVC%,PEF,MVV,Cdyn)of rats in all groups were observed,and the rats’ middle lobe were taken for pathological observation.The effects of Shengxian Tongluo decoction and N-acetylcysteine on alveolitis and pulmonary fibrosis in rats with pulmonary fibrosis were observed by HE staining.The levels of mTOR,p70S6K,MMP2,TIMP1,Col Type Ⅰ,Col Type Ⅲ,Col Type Ⅳ,HA,FN protein and gene expression in lung tissue of each group were detected by Western blotting and RT-PCR.Results:The lung function demonstrate that Shengxian Tongluo decoction can improve the cdyn index of rats(P<0.05),but for some indexes of lung function,FVC,FEVO.3/FVC,PEF,MVV,the improvement is not obvious(P>0.05);Pathological results show that the model group rats have alveolar structure damage,interstitial fibers hyperplasia,a large number of inflammation cells infiltration,Shengxian Tongluo decoction group and N-acetylcysteine group compared with model group are alleviated.The results of Western blotting and RT-PCR show that the ECM proteins(type I collagen,type Ⅲ collagen,type Ⅳ collagen,HA,FN)and mTOR/p70S6K signaling pathways,MMPs/TIMPs protease system relative expression of mRNA of the lung tissue of rats in the group of Shengxian Tongluo decoction were lower than those of the model group(P<0.01),and the protein level of ECM protein,mTOR/p70S6K signal pathway and MMPs/TIMPs system in the Shengxian Tongluo decoction 3rd day intervention group was also lower than that in the model group(P<0.01).The relative mRNA expressions of HA in Shengxian Tongluo decoction 3rd day and 14th day intervention groups were lower than that in the corresponding western medicine groups(P<0.05 or P<0.01).The relative mRNA expressions of Col Type Ⅰ and FN in Shengxian Tongluo decoction 14th day and 28th day intervention groups were significantly lower than that of the corresponding western medicine groups(P<0.05 or P<0.01).The relative mRNA expressions of MMP2 and TIMP1 in the Shengxian Tongluo decoction 3rd day intervention group were lower than that on the 14th day and the 28th day intervention groups(P<0.01).Conclusion:Shengxian Tongluo decoction can inhibit the deposition of ECM by regulating mTOR/p70S6K signaling pathway,MMPS/TIMPs system and other related mechanisms.And Shengxian Tongluo decoction can also reduce the inflammatory cell infiltration,fibrous tissue hyperplasia,degree of pulmonary fibrosis and decline of Cdyn.