苦参碱诱导大鼠肝卵圆细胞表型改变

背景与目的:在肝细胞癌发生过程中出现卵圆细胞不典型增生,阻断其恶变或诱导其向肝细胞方向分化是肝癌化学预防的重要途径,为寻找有效的卵圆细胞分化诱导药物,我们建立了大鼠卵圆细胞增殖模型,研究苦参碱对化学诱发肝癌模型中卵圆细胞表型的改变。方法:SD大鼠喂饲2-乙酰氨基芴加三分之二肝切除术建立卵圆细胞增殖模型,模型组、低剂量苦参碱组、高剂量苦参碱组、对照组,光镜、透射电镜观察卵圆细胞的超微结构;免疫组织化学方法检测造血干细胞标志Thy-1、甲胎蛋白(alphafetoprotein,AFP)表达的变化;酶组织化学方法检测γ-谷氨酰转肽酶(γ-glutamyltranspeptidase,γ-GT酶)、三磷酸腺苷酶(adenosinetriphosphatase,ATP酶)表达的变化。结果:超微结构显示高剂量苦参碱组卵圆细胞体积较模型组大,含有更多的粗面内质网等细胞器。Thy-1免疫组化表达指数模型组为8.15±2.64,低剂量苦参碱组为5.27±1.32,高剂量苦参碱组为3.83±0.35,对照组为1.63±0.22,高剂量苦参碱组显著低于模型组(P<0.05);AFP阳性细胞计数模型组为15.36±4.42,低剂量苦参碱组为9.75±2.41,高剂量苦参碱组为7.33±1.38,对照组为2.51±0.93,高剂量苦参碱组显著低于模型组(P<0.05);低剂量苦参碱对γ-GT阳性病灶抑制率为33.35%,高剂量苦参碱抑制率为55.37%,高剂量苦参碱组显著高于低剂量苦参碱组(P<0.05)。结论:苦参碱抑制卵圆细胞增生,使卵圆细胞的超微结构发生改变,降低Thy-1、AFP、γ-GT酶表达,表明苦参碱可诱导卵圆细胞表型改变。

Inducement effect of matrine on phenotype changes of rat hepatic oval cells

BACKGROUND & OBJECTIVE: Atypical dysplasia of oval cells plays a crucial role in the pathogenesis of hepatocellular carcinoma (HCC). To prevent oval cells from carcinogenesis or induce them to differenciate into hepatocytes will become an effective way for chemical treatment of HCC. This study was to construct a rat hepatic oval cell proliferation model, and observe the inducement effect of matrine on phenotype changes of hepatic oval cells. METHODS: Hepatic oval cell proliferation model was constructed with SD rats by 2-acetaminofluorene administration and 2/3 partial hepatectomy, and divided into model group, low dose matrine group, high dose matrine group, and control group. Ultrastructure of oval cells was observed by electron microscopy. The expression of Thy-1 and alpha fetoprotein (AFP) was detected by immunohistochemistry. The expression of gamma-glutamyl transpeptidase (gamma-GT) and adenosine triphosphatase was detected by enzyme-histochemisty. RESULTS: The oval cells in high dose matrine groups were larger and contained more rough endoplasmic reticula than those in control group. The expression index of Thy-1 was 8.15+/-2.64 in model group, 5.27+/-1.32 in low dose matrine group, 3.83+/-0.35 in high dose matrine group, and 1.63+/-0.22 in control group; it was significantly lower in high dose matrine group than in model group (P<0.05). The number of AFP-positive cells was 15.36+/-4.42 in model group, 9.75+/-2.41 in low dose matrine group, 7.33+/-1.38 in high dose matrine group, and 2.51+/-0.93 in control group; it was significantly lower in high dose matrine group than in model group (P<0.05). The inhibition rate of gamma-GT was significantly higher in high dose matrine group than in low dose matrine group (55.37% vs. 33.35%, P<0.05). CONCLUSION: Matrine can inhibit the proliferation of oval cells, induce ultrastructure changes, and suppress the expression of Thy-1, AFP, and gamma-GT.