The Inhibitory Effect And Mechanism Of Anlotinib On Mouse Model Of Pulmonary Fibrosis

BackgroundPulmonary fibrosis is one of the more serious interstitial pulmonary diseases.Compared with other respiratory diseases,such as asthma and chronic obstructive pulmonary disease(COPD),IPF remains poorly understood among clinicians.In fact,IPF has a very high morbidity and mortality rate,with a very low5-year survival rate.The current status of treatment for IPF is not encouraging.Only pirfenidone and nidanib have been publicly approved and marketed in the country.Existing literature has shown that pirfenidone and nidanib can play a very good role in improving the lung function of patients.However,the cost of these two drugs is very high,limiting clinical use.However,stem cell technology in the treatment of pulmonary fibrosis is not mature enough,and there is still a long way to go before it can be applied in clinical practice.Problems such as poor quality of donor lung and acute or chronic rejection reaction after surgery have not been solved.Therefore,frankly speaking,the treatment of IPF is still in a dilemma.Anlotinib is an anti-cancer drug,consider to Anlotinib for both drugs and manes nidanib,cloth similar mechanism of action,the hypothesis of this study is that Anlotinib ROM for can prevent and treat animal model of pulmonary fibrosis in mice is shown to improve lung function,improve lung fibrosis,pathology,changes of pulmonary fibrosis related marker.The results of this study,if consistent with the study hypothesis,will be able to lay a preclinical basis for the expanded clinical indication of amrotinib for the treatment of IPF,which is conducive to the Phase II and III clinical studies of amrotinib for the treatment of IPF.ObjectiveTo investigate whether Anlotinib could inhibit bleomycin(BLM)-induced pulmonary fibrosis in mice and its mechanism.MethodsThe pulmonary fibrosis model was induced by bleomycin(BLM)nasal drops,and the intervention was carried out with phosphate buffer solution(PBS)and different doses of amlotinib by gavage.Blank control group was treated with PBS nasal drip and PBS gavage.Male C57/BL6 mice were randomly divided into 5 groups: PBS+PBS group(PP group),BLM+PBS group(BP group),BLM+ amlotinib 1.5mg·kg-1 group(BA1.5 mg·kg-1),BLM+ amlotinib 3.0 mg·kg-1 group(BA3.0 mg·kg-1),and BLM+amlotinib 6.0 mg·kg-1 group(BA6.0 mg·kg-1).Histopathological analysis was performed to detect MDA and HYP contents,ELISA was used to detect inflammatory cytokines in BALF,quantitative PCR was used to detect cytokines in lung tissues,and Western blot was used to analyze the expression levels of TGF-β1,mitochondrial division/fusion proteins OPA1,Mfn2 and DRP1,MFF and epithelial mesenchymal transformation(EMT)markers.ResultsCompared with PP group of mice,BP group integral increase lung inflammation in mice,increased pulmonary fibrosis scores,bronchoalveolar lavage fluid(BALF),malondialdehyde content,achievement of lung tissue amino acid content increased,elevated inflammatory factor levels in BALF,lung tissue inflammation factor m RNA levels,TGF-beta 1 protein increases,E-Cadherin protein decreased,while Vimentin and N-Cadherin protein increases,OPA1 and MFN-2 protein is reduced,and DRP1 and MFF protein increased.6.0mg·kg-1 amlotinib intervention inhibited pulmonary inflammation score,pulmonary fibrosis score,BALF malondialdehyde content,lung tissue hydroxyproline acid content,BALF inflammatory factor level,lung tissue inflammatory factor m RNA level,TGF-β1,DRP1 protein expression and Vimentin protein expression,and increased the expression of MFN-2 and E-cadherin protein.In addition,3.0mg·kg-1 amlotinib intervention inhibited MFF protein expression.3.0mg·kg-1 and 6.0mg·kg-1 amlotinib inhibited N-cadherin protein expression and increased OPA1 protein expression.Conclusions Anlotinib inhibited bleomycin-induced pulmonary fibrosis in mice,and the mechanism is related to inhibition of pulmonary inflammation,oxidative stress,TGF-β1expression,mitochondrial mitosis/fusion,and EMT.