Effects Nrf2of Matrine In The Pulmonary Fibrosis Rats On Expression Of γ-GCS In Human Fetal Lung Fibroblast

Background:Oxidant/antioxidant imbalance play an important role in the pathogenesis ofpulmonary fibrosis.γ-glutamyl cysteine synthetase (γ-glutamylcysteine synthetase,γ-GCS) is the rate-limiting enzyme of glutathione (glutathione,GSH) synthesis.It was considered as very important antioxidant enzyme.Studies invitro suggested that nuclear factor erythroid-derived-associated factor-2(NF-E2related factor2,Nrf2) controll the expression of anti-oxidase.Studies sugesstedMatrine has the effect of anti-fibrosis.And matrine has the effect of anti-fibrosis.Objectives:Studies in vitro human fetal lung fibroblast suggested that nuclear factor erythroid derived associated factor-2(NF-E2related factor2,Nrf2) controll the expression of anti-oxidase.Studies sugessted Matrine has the effect of anti-fibrosis.And matrine has the effect of anti-fibrosis.ABSTRACTBackground:Oxidant/antioxidant imbalance play an important role in the pathogenesis ofpulmonary fibrosis.γ-glutamyl cysteine synthetase (γ-glutamylcysteine synthetase,γ-GCS) is the rate-limiting enzyme of glutathione (glutathione,GSH) synthesis.It was considered as very important antioxidant enzyme.Studies invitro suggested that nuclear factor erythroid-derived-associated factor-2(NF-E2related factor2,Nrf2) controll the expression of anti-oxidase.Studies sugesstedMatrine has the effect of anti-fibrosis.And matrine has the effect of anti-fibrosis.Objectives:Studies in vitro human fetal lung fibroblast suggested that nuclear factor erythroid derived associated factor-2(NF-E2related factor2,Nrf2) c ontroll the expression of anti-oxidase.Studies sugessted Matrine has the effect of anti-fibrosis.And matrine has the effect of anti-fibrosis.Methods: In vitro human fetal lung fibroblast(MRC-5) cultured to theexponential growth phase with different concentration of erythromycin, cells culturedwith MEM/NEAA; cells cultured with matrine(Mat);then were randomly divided intocontrol group (groupA),Bleomycin group (0.25mmol/L Mat group B、0.5mmol/LMatgroupC、1.0mmol/LMat groupD), Added into SiRNAgroup (0.25mmol/LMatgroupE、0.5mmol/LMat groupF、1.0mmol/LMat groupG), All of the group cells werecultured1hours、2hours、4hours and8hours, Reverse transcription-polymerase chainreaction(RT-PCR) measured the expression of Nrf2and γ-GCS mRNA in vitro humanfetal lung fibroblast(MRC-5).Western blot measured the protein expressions of Nrf2and γ-GCS.Results:(1)Contrast to group A,The protein expression of γ-GCS and nrf2weresignificantly higher in the in the MRC-5else groups. Group E、F、Gwas lower thanGroup B、C and D(P<0.01).(2) Contrast to group A,The expression of γ-GCS and nrf2mRNA weresignificantly higher in the lung tissue in the else groups.Group B was lower thanGroup C,D and E(P<0.01).Contrast to group B、C、D,Group E、F、G was lower thanBleomycin groups， and was lower than GroupA(P<0.01).(3)They were positively correlated between the expression of γ-GCS protein andthe expression of Nrf2protein,mRNAConclusions:By controlling the expression of Nrf2to indirectly control the expression ofγ-GCS. This is a possible mechanism why matrine inhibit pulmonary fibrosis in.