Protective Effects Of Neferine On Amiodarone-induced Pulmonary Fibrosis In Mice

Objectives To explore the protective effects of Neferine on Amiodarone-inducedpulmonary fibrosis in mice.Methods Mice were randomized into5groups as control,model,neferine, predniso-lone,pirfenidone. Control group,which received three intratracheal instillation of sa-line on the1st、3rd and5th day. Other groups received three intratracheal instillationof amiodarone to established the model of pulmonary fibrosis. From day6,control andmodel groups received saline10ml/kg,neferin e,prednisolone,pirfenidone groups re-ceived neferine20mg/kg,prednisolone15mg/kg,pirfenidone100mg/kg intragastricinjection twice a day respectively. Animals were sacrificed on the21st day. Recordedthe body weight at the same period during the experiment. Lungs were isolated tomeasure lung index (lung wet weight in milligrams versus body weight ingrams),Hematoxylin-eosin and Masson’s trichrome for evaluating alveolitis and fibro-sis. Chloramine-T method was used to assay the content of hydroxyproline. PlasmaSurfactant Protein(SP)D was assayed by ELISA,interferon-γ(IFN-γ)and interleu-kin-4(IL-4) levels were assayed with a sandwich ELISA technique to establish theTh1/Th2balance. Flow Cytometry was used to assess the percentage ofCD4~+CD25~+Tregs/cells in CD4~+cells.Results All groups’ body weight were gradual recovery from day7,Neferine signif-icantly restored reductions in body weights associated with amiodarone.Model grouplung index and content of hydroxyproline were increased compared with controlgroup,neferine inhibited lung index and content of hydroxyproline when comparedwith model group. Model lung histopathology revealed massive infiltration of in- flammatory cells and fibrosis,neferine significantly improved this. Neferine-treatedmice showed a down modulation of SP-D when compared with model (23.57±2.14ng/ml vs44.00±1.80ng/ml,p<0.05).Compared with model group,neferine increasedcontent of IFN-γ while decreased IL-4,restoring Th1/Th2balance. Neferine inhibitedthe trend of increasing proportion of CD4~+CD25~+Tregs in CD4~+cells when comparedwith model group (3.92±0.13%vs5.26±0.14%,P<0.05).Conclusion Protective effects of neferine on amiodarone-induced pulmonary fibrosisin mice probably through decreasing the proportion of CD4~+CD25~+Tregs inCD4~+cells,restoring Th1/Th2balance and inhibiting SP-D level. Objectives To study the proliferation effects of Amiodarone with different concentra-tion and action time on lung fibroblasts (LF).Methods LF cells in culture with10%bovine serum in logarithm growth stageswere planted in96well plates. LF cells were treated with Amiodarone of differentconcentration (0.1μmol/L~100μmol/L),at12,24,48h after administering Amioda-rone,the proliferation of LF cells was measured by MTT colorimetric assay.The effectof different concentration (1μmol/L~100μmol/L) neferine on amiodarone-inducedproliferation of LF were tested by MTT.Results There is significant effect of proliferation on LF cells(P<0.05)compareswith control group when the cells were created with Amiodarone of10μmol/L on12,24,48h,anthe10μmol/L neferine inhibit the growth of LF by amiodarone and theother concentrations have no effect.Conclusion Amiodarone10μmol/L can promote the proliferation of LF cells at12,24,48h,and at48h,the rate of proliferation is more significant.10μmol/L ne-ferine significant inhibit the proliferation of LF cells by amiodarone.