The Study Of Effect And Mechanism Of Nervilia Fordii Against Lung Fibrosis

The purpose of study was to explore Herba Nervilia fordii,the leaves or the whole plant of Nervilia fordii(Hance)Schltr.(Orchidaceae)induced effect and mechanism against lung fibrosis by the signing pathway of TGF-β/Smad,CTGF and TGF-β/ERK.The main content include the model of lung fibrosis was replicated by BLM;To observe acute toxicity of NFTF;By applicatting LC-MS method to detect ComplanatUside and it,s metablish in the blood ang lung tissues by oral complanatuside;To observe interventive effect and mechanism of NFTF on experiental rat model of lung fibrosis by BLM;To observe effect and mechanism of NF-1 and NF-2 on experiental of 3T6 Swiss albino;To observe interventive effect and mechanism of NF-1 on experiental rat model of lung fibrosis by BLM.The results showed that to replicate Pulmonary fibrosis model experiment by BLM in rats,the dose of BLM in 4,5 mg/kg can lead to the formation of pulmonary fibrosis in lung tissue of rats after 14 days.The primary pulmonary fibrosis model that was formed after 14 days thougan sigle administration 4 mg/kg of BLM ean be used as this topic subsequent experiment.In NFTF maximum dosage experiment,NFTF maximum dosage is 2.448 g,with 1/10 of the maximum dosage as for NFTF effective dose,Meanwhile combined with the conversion of clinical drug dose,to get NFTF pharmacodynamics of high,medium and low dose respectively are 400,200 and 100 mg/kg.Complanatuside pharmacokinetic experiments in vivo,mice were gavaged at a dose of 144 mg/kg of Complanatuside,biological samples in different time were taken before treatment,mouse biological samples were analyzed with the established methods,Complanatuside and its metabolites are consistent with non compartmental model by DAS3.0 fitting pharmacokinetics dynamic.The blood drug concentration,residence time as well as the apparent volume is the largest in serum and tissues.The original drug and its metabolite pharmacokinetic parameters are analyzed in plasma and lung tissue,The results show that drug concentration in lung tissues with the peak time delay are compared with plasma,and the peak concentration in lung tissues is lower than the peak concentration of the drug in the plasma.At the same time,the apparent volume of distribution of rhamnocitrin in the lung tissues is similar in plasma,the average dwell time in the lung tissues is relatively long than in the plasma.In the experiment that NFTF affects the BLM induced pulmonary fibrosis model,After NFTF drug intervention,compared with model group,the HYP content in serum and lung tissue of rats decreased significantly,the oxidation and anti-oxidation function index T-AOC,GSH were rebounded,MPO content significantly reduced to normal]evels(P<0.05);In the drug group,pathology was observed to have a small amount of inflarnation factor invasion in lung tissue,less fibroblasts proliferation and collagen deposition near the Iung and bronchus were decreased.The group of NFTF high were effective obviously.From the point of gene and protein level,other drugs group can cut or increase the contents of relative protein in pathway of TGF-β/Smad and TGF-p/ERK.All statistically significant difference(P<0.05).In the experiment that NF-1 and NF-2 affects the TGF-β induced embryonic fibroblast(3T6 swiss ablino)proliferation experiment,compared with model group,NF-1 and NF-2 can significantly reduce the contents of collagen type Ⅰ and Ⅲ,TNF-α and IL-1β that rised in cell supernatant fluid due to TGF-β.The content of collagen deposition is given priority to with collagen type 1.At the same time,NF-1 and NF-2 can increase SOD and GSH content(P<0.05).From the point of gene and protein level,drug group can cut or increase the contents of relative protein in pathway of TGF-β/Smad and TGF-β/ERK.All statistically significant difference(P<0.05)In the experiment that NF-1 affects the BLM induced pulmonary fibrosis model,After NF-1 drug intervention,compared with model group,the HYP content in serum and lung tissue of rats decreased significantly,the oxidation and anti-oxidation function index T-AOC,GSH were rebounded,MPO,TNF-α and IL-1β content significantly reduced to normal levels(P<0.05);In the drug group,pathology was observed to have a small amount of inflammation factor invasion in lung tissue,less fibroblasts proliferation and collagen deposition near the lung and bronchus were decreased.From the point of gene and protein level,drug group can cut or increase the contents of relative protein in pathway of TGF-β/Smad and TGF-β/ERK.All statistically significant difference(P<0.05).In this paper,the model of pulmonary fibrosis induced by BLM in rats,NFTF and its main active components of NF-1 and NF-2 can significantly improve the content of serum and lung tissue in HYP,T-AOC,MPO,GSH,TNF-α and IL-1β and other biochemical indicators,inhibit the proliferation of fibroblasts,fibroblast cell danage protection,at the same time can significantly reduce the collagen and extracellular matrix near the hilum of lung and bronchial lung tissues(α-SMA)deposition,thereby reducing lung fibrosis in rats.The mechanism of anti fibrosis drug may be through inhibiting the expression of TGF-βexpression,further regulation of Smad and ERK genes and protein downstream,reduce inflammation,antioxidant index and collagen expression of the extracellular matrix,thereby preventing the formation of rat pulmonary fibrosis.