细脚拟青霉多糖对肝星状细胞体外活化的影响

Effect of Paecilomyces tenuipes polysaccharides on hepatstellate cell in vitro

目的探讨细脚拟青霉多糖(PtPs)对原代分离培养的新生大鼠肝星状细胞(HSC)活化的影响。方法将不同浓度的PtPs作用于新生大鼠的HSC,反转录聚合酶链反应(RT-PCR)检测HSC中转化生长因子β1(TGF-β1)mRNA和白介素10(IL-10)mRNA的表达,细胞免疫组织化学检测在不同培养时间HSC中α-平滑肌肌动蛋白(SMA-α)、细胞角蛋白18(CK-18)和Ⅰ型胶原的表达,检测单胺氧化酶(MAO)在HSC的培养上清和线粒体中的活性,检测羟脯氨酸(Hyp)在HSC的培养上清和胞浆中的含量。结果与对照组比较,各浓度PtPs组TGF-β1mRNA的表达均显著增加(P<0.01),IL-10mRNA的表达均显著下降(P<0.01);随着HSC培养时间的延长,PtPs各组较对照组CK-18的表达下降,SMA-α和Ⅰ型胶原的表达增强;与对照组比较PtPs 250 mg/L组培养上清和线粒体中的MAO活性最高(P<0.01),PtPs 62.5 mg/L组胞浆中Hyp含量最高(P<0.05)。结论PtPs能促进新生大鼠HSC体外活化。

Purpose To investigate the effect of different concentfitions of Paecilomyces tenuipesl polysaccharides（PtPs） on hepat-stellate cell （HSC）. Methods To choose different concentritions of PtPs on HSC through proliferation assay. Expression of transforming growth factor β1 （TGF-β1）mRNA and interleukin（IL-10） mRNA of HSC was detectd by reverse transcriptase-polymerase chain reaction（RT-PCR） . Growth and decline of α-smooth muscle actin （SMA-α）, cytokeratin 18 （CK-18）and Collagen Ⅰ were observed by immunohistochemistry method. Activity of monoaminoxidase （ MAO ） and content of hydroxyproline （Hyp） were detectd through zymochemistry method. Results The results showed that expression of TGF-β1 mRNA significantly increased（ P 〈 0.01 ） in PtPs groups vs PtPs 0 mg/L group and expression of IL-10 mRNA decreased （ P 〈 0.01 ） ; along with HSC＇s culture time expression of SMA-α and Collagen Ⅰ increased in PtPs groups and CK-18 decreased; MAO activity of PtPs 250 mg/L group in supernatant and mitochondria was the highest vs PtPs 0 mg/L group （P 〈 0.01 ）, the same of Hyp contentent of PtPs 62.5 mg/L group in endochylema （ P 〈 0.05 ）. Conclusion PtPs can promote HSC proliferation and activation without exact dose-effect relationship in vitro.