1.Medical Research Center,Mudanjiang Medical College,Heilongjiang Province,Mudanjiang 157011,China;
2.Inspection and Legal Affairs Section,Mudanjiang Public Health Inspection Institution,Heilongjiang Province,Mudanjiang 157011,China
Abstract：ObjectiveTo preliminarily study the influence of recombinant protein of latency-associated peptide(LAP)of recombinant truncated TGF-β precursor on proliferation of hepatic stellate cells (HSC)-T6.MethodsThe truncated LAP fragment was amplified by polymerase chain reaction(PCR)and the prokaryotic expression vector was constructed.After double enzyme digestion and DNA sequencing,the protein expression was induced by IPTG at 0.5 mmol/L at 37°C for 4 h.The expressed protein was detected by SDS-PAGE and Western blot,after that the recombinant protein was by nickel agarose affinity chromatography for purification.The recombinant protein was applied to rat hepatic stellate cell-T6 (HSC-T6),and the influence of different concentrations of recombinant protein on cell proliferation was detected by MTT assay.ResultsThe prokaryotic expression vector was successfully constructed and the recombinant protein was expressed and purified successfully.The proliferation of HSC-T6 cells decreased with the increase of the recombinant protein concentration.ConclusionThe recombinant protein inhibits the proliferation of HSC-T6 cells,and lays the foundation for the subsequent study on the interference of LAP fragments on the activation of TGF-β and the influence on organ fibrosis.
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