【摘 要】
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Aim:Oxymatrine(OM)is a monomer extracted from Sophora species that has been used to treat fibrotic diseases.While its anti-fibrotic potential in clinical application is limited due to a lack of specif
【机 构】
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Department of Pharmacy,The Third Xiangya Hospital,Central South University,Changsha,Hunan,China
【出 处】
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2017年中国药学大会暨第十七届中国药师周
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Aim:Oxymatrine(OM)is a monomer extracted from Sophora species that has been used to treat fibrotic diseases.While its anti-fibrotic potential in clinical application is limited due to a lack of specific mechanism,so our study aim to explore the molecular mechanism of inhibitory effect of Oxymatrine on HSCs activation and and its modulation on the TGF-β/Smad signaling pathway.METHODS:HSC-T6 was cultivated for our study,the effect of different concentrations and time of OM on HSC-T6 cell viability was determined by MTT assay.Then groups were divided as follows,(1)Control group,(2)TGF-β1 group,(3)125 μg/mL OM+TGF-β1 group,(4)250 μg/mL OM+TGF-β1 group,(5)500 μg/mL OM+TGF-β1 group,(6)IFN-γ+TGF-β1 group,(7)miR-195 mimic+OM+TGF-β1 group,(8)miR-195 mimic control+OM+TGF-β1 group.HSC-T6 were respectively transfected 100 nM miR-195 mimic/mimic control and incubated for 24 h,then the drug treatment group was treated with 250 μg/mL OM for 1 h,while the positive drug group was treated with 500 U/mL IFN-γ for 1 h,at last added 5 ng/mL TGF-β1 to the cells,cultured for 24 h.The mRNA expression of miR-195,Smad7 and HSC-T6 activation marker gene α-SMA were detected by Quantitative Real-time PCR.The protein expression of α-SMA were detected by Western Blot.Results:Inhibition effect of OM on HSC-T6 cell proliferation was strengthened along with the elevated concentrations and prolonged incubation time.OM could decrease cell activation induced by TGF-β1 in a dose-dependent method,reduce the expression of miR-195 and α-SMA(P<0.01),and promote the expression of Smad7(P<0.05).OM and IFN-γ had similar anti-activation effect on HSC-T6.HSC-T6 transfected with miR-195 mimic and pretreated OM could upregulate the expression of miR-195 and α-SMA(P<0.05),and downregulate the expression of Smad7(P<0.05)compared with the OM group.Conclusions:OM could attenuate HSCs activation induced by TGF-β1 in a dose-dependent method,which achieved via down-regulating the expression of miR-195 and up-regulating Smad7 which is the target of miR-195.
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