【作者】 刘东；

【导师】 陈志良；

【作者基本信息】 南方医科大学 ， 药理学， 2007， 博士

【摘要】 目的：建立小鼠和大鼠急、慢性肝损伤的模型，研究甘草甜素与苦参碱联合使用对肝纤维化以及大鼠肝星状细胞的影响，进而观察甘草甜素与苦参碱联合用药对肝纤维化的防治作用并初步探讨其作用机制。方法：（1）采用四氯化碳（CCl₄）制备小鼠的急性、慢性肝损伤模型，通过血清酶学指标和组织病理学观察不同剂量的甘草甜素与苦参碱合剂对肝纤维化的保护作用。（2）采用CCl₄诱导大鼠肝纤维化模型，通过测定大鼠血清中Ⅲ型前胶原（PC-Ⅲ）、层粘蛋白（LN）、透明质酸（HA），并作肝组织病理学形态分析研究不同剂量合剂治疗组的效果。（3）以培养的大鼠肝星状细胞（HSC）为实验对象，观察单用甘草甜素或苦参碱以及联合用药对活化HSC分泌Ⅰ型胶原、HA和LN的影响；同时研究甘草甜素与苦参碱联合使用对Fe-NTA致HSC-T6细胞氧应激后的细胞增殖、Ⅰ型胶原水平、MDA、GSH、SOD及GSH-PX活性的影响。结果：（1）在小鼠急、慢性肝损伤模型的基础之上，甘草甜素与苦参碱合剂组均可降低血清中ALT、AST、LDH、ALP的活性，缓解汇管区的纤维组织增生。（2）合剂低、中、高各治疗组均能显著改善肝功能，不同程度的降低PC-Ⅲ、LN、HA水平，组织学检查也显示具有抗纤维化作用。（3）研究结果显示：在无包被的塑料培养皿诱导的肝纤维化细胞模型中，与空白对照组比较，1～20μmol／L浓度范围的甘草甜素，1～20μmol／L浓度范围的苦参碱均可剂量依赖地抑制HSC的增殖，并不同程度抑制Ⅰ型胶原、HA和LN分泌，综合比较各治疗组对HSC分泌Ⅰ型胶原、HA和LN的影响，合用组优于单用组，尤以甘草甜素与苦参碱合用组效果最佳。在Fe-NTA诱导氧应激模型中，与模型组比较，甘草甜素与苦参碱均可抑制HSC的增殖，其中甘草甜素与苦参碱合用优于单用（有显著性差异），与阳性对照组效果相当。各治疗组还可降低HSC培养液中Ⅰ型胶原的水平，尤以甘草甜素与苦参碱合用组Ⅰ型胶原水平最低（有显著性差异）。结论：（1）低、中、高合剂组对小鼠急性肝损伤有很好的保护作用，表现在均能抑制肝酶（ALT、AST、LDH、ALP）的上升，可抑制胶原的生成和沉积。（2）合剂低、中、高各治疗组均能显著改善肝功能，降低PC-Ⅲ、LN、HA水平，组织学检查也显示具有抗纤维化作用。（3）甘草甜素与苦参碱合剂可显著抑制HSC的增殖及ECM分泌，且合用优于单用。更多还原

【Abstract】 Objective: To investigate the influence of glycyrrhizin plus oxymatrineon glycyrrhizin plus oxymatrine, acute and chronic liver fibrosis modes were set upto study the prevention and treatment of glycyrrhizin plus oxymatrine, additionallly,we carried out a profound investigation into the mechanism involved in the effectand influence on the hepatic stellate cells （HSC）.Methods: （1） The acute and chronic liver fiborsis models of mice inducedby carbon tetrachloride （CCl₄） were designed to investigate the protective effects bymeasuring the lever of serum enzymes and examing histological morphous. （2） Ratsliver fiborsis model was induced by carbon tetrachloride （CCl₄）, the level ofserum procollgan type-Ⅲ（PC-Ⅲ）, hyaluronic acid （HA）, laminin （LN）, liverfunction and liver histological examination were measured respectively. （3） HSC ofrats were cultured to observe the action of glycyrrhizin, oxymatrine and mixture onthe proliferation and collagenⅠ, and then investigate the synergism effect on HSC.Furthermore, we investegated the effecte of glycyrrhizin plus oxymatrine on theactivity of MDA、GSH、SOD and GSH-PX in the supernatant of cell culturefollowed by treatment of Fe-NTA.Results: （1） Glycyrrhizin plus oxymatrine ameliorated the level of serumALT, AST, DH, ALP in the models of CCl₄-induced liver fibrosis significantly. （2）The lever of serum PC-Ⅲ, HA, LN decreased and liver function were improved. The histological examination also demonstrated its anti-fibrotic effect. （3） The resultsshows that compared with the contol group, 1～20μmol/L glycyrrhizin and 1～20μmol/L oxymatrine can inhibit the proliferation of HSC dose-dependly and inhibitthe secretion of PC-Ⅲ, HA, LN, compared all groups comprehensivly, we could alsoconcluded that the the effect of combination group is than the single use group,especially the combine use of glycyrrhizin plus oxymatrine.Conclusion: Glycyrrhizin plus oxymatrine showed significant reductionsin the ALT、AST、LDH and ALP activities, and inhibited the formation anddeposition of collagen. The histological examination demonstrated the anti-fibroticeffect of mixtura through ameliorating the activity of serum PC-Ⅲ, HA, LN. In vitro,the mixtura inhibited the secreation of ECM and also had anti-proliferation effect onHSC. Furthermore, we observed the anti-fibrotic effect of mixtura was better thaneither glycyrrhizin or oxymatrine alone.更多还原