细脚拟青霉多糖对乙醇诱导L-02细胞损伤的保护作用研究

 目的探讨细脚拟青霉多糖(PtPs)对乙醇诱导肝细胞损伤的体外保护作用。方法采用RT-PCR分别检测乙醇诱导损伤的乙醇组、PtPs处理组(PtPs+乙醇)及正常对照组L-02细胞肿瘤坏死因子-α(TNF-α)mRNA和热休克蛋白90(HSP90)mRNA的表达;检测细胞培养上清液中黄嘌呤氧化酶(XOD)、超氧化物歧化酶(SOD)、乳酸脱氢酶(LDH)活性及丙二醛(MDA)含量;检测细胞匀浆中三磷酸腺苷酶(ATPase)活性,并对各组细胞进行Annexin-FITC标记染色观察凋亡细胞情况。结果与乙醇组比较,乙醇+PtPs组细胞内TNF-α mRNA表达下降(P<0.01),HSP90 mRNA表达增加(P<0.05或P<0.01);细胞培养上清液中SOD活性升高、XOD及LDH活性下降、MDA含量下降(P<0.05或P<0.01);细胞匀浆中ATPase活性上升(P<0.05或P<0.01);PtPs处理后可见乙醇诱导的L-02细胞凋亡和坏死被明显抑制。结论PtPs对乙醇诱导肝细胞的损伤具有保护作用。

Study of the Hepatoprotective Effect of Paecilomyces tenuipes Polysaccharides on Alcohol-Induced L-02 Cell Injury

OBJECTIVE To investigate the protective effect of Paecilomyces tenuipes polysaccharides(PtPs)on alcohol-induced hepatocyte injury in vitro.METHODS RT-PCR was used to detect the expressions of tumor necrosis factor-α(TNF-α)mRNA and heat shock protein 90(HSP90)mRNA in L-02 cells of alcohol group,PtPs disposed groups(PtPs+ alcohol)and normal control group. The levels of xanthinoxidase(XOD),superoxide dismutase(SOD),lactic dehydrogenase(LDH),malondialdehyde(MDA)in supernatant and adenosine triphosphatase(ATPase)in cell homogenate were determined. The cells in each group were stained by Annexin-FITC and the apoptosis was observed with fluorescent microscope.RESULTS Compared with those in the alcohol group,the expression of TNF-α mRNA was decreased(P<0.01) and that of HSP90 mRNA was increased(P<0.01 or P<0.05) in PtPs disposed groups. The activity of SOD increased and the levels of XOD,LDH,MDA decreased in supernatant of PtPs disposed groups(P<0.01 or P<0.05). The activity of ATPase in cell homogenate increased (P<0.01or P<0.05). PtPs significantly inhibited apoptosis and death of L-02 cells induced by alcohol. CONCLUSION PtPs at a certain concentration could protect hepatocyte from the injury induced by alcohol.