槲芪散阻断肝癌前病变的机制研究

Mechanism study of Huqisan on prevention and cure hepatic precancerosis

目的:探讨槲芪散及其拆方阻断肝癌前病变的作用。方法:用二乙基亚硝胺(DEN)建立Solt-Farber二步大鼠肝癌前短期实验模型,用槲芪散全方(Ⅰ号方)及槲芪散拆方(Ⅱ号方)进行干预,应用流式细胞术(FCM)测定肝癌前病变大鼠肝组织的DNA相对含量;观察肝组织病理及超微结构的变化。采用组织化学染色法检测肝癌前病变组织中γ-谷氨酰转肽酶(γ-GT)、葡萄糖-6-磷酸脱氢酶(G-6-Pase)、三磷酸腺苷酶(ATPase)的活性。采用琼脂糖电泳法测定各组大鼠血清中乳酸脱氢酶(LDH)总活力及其同功酶、血清γ-谷氨酰转肽酶同功酶Ⅱ(γ-GT-Ⅱ)。结果:槲芪散Ⅰ号方可减少大鼠肝癌前病变部位的γ-GT灶的发生数和灶面积,血清中γ-GT-Ⅱ阳性率明显下降。DI接近于正常肝细胞;可将调节能量代谢的酶类基本维持在正常水平,ATPase与G-6-Pase增生灶呈阳性反应。结论:槲芪散具有明显阻断肝癌前病变的作用,而槲芪散拆方无此作用。

Objective： To investigate the obstructive effect of Huqisan and break this prescription of hepatocarcinogenesis. Methods： A Soh-Farber two-step test model of prehepatocarcinoma was established by diethylnitrusamine （DEN） and 2-acetylaminofluorene （AAF） in rats. Complete prescription-Huqisan, also called drug No. Ⅰand disassemble this prescription, also called drug No.Ⅱ to carry out intervention. DNA contents of rat hepatic tissues were measured with flow cytometer （ FCM ）, and observe the proliferation cycle of cell transmingration vary with Huqisan and its disassemble prescirption. Heterotypic proliferative focus of hepatic cells were reviewed by light microscopy and electron microscopy. γ-glutamy-transpeptidase-isoenzyme （γ-GTase）, adenosin triphosphatase （ATPase）, glucose-6-phosphat dehydrogenase （G-6-PD） were determined with histochemical methods in rats. Serum lactic dehydrogenase （LDH） and its isoenzyme, serum γ-glutamy-transpeptidase-isoenzyme- Ⅱ （γ-GT- Ⅱ ） were determined with the way of agarose electrophoresis in rats. Results： The results showed the focal amount （ numbers/cm^2 ） of γ-GTase abnormal elevate in model group, but the rats were treated with complete prescription-Huqisan, the activities these enzymes descend obviously, P 〈 0. 005 ; the inhibitive rate was 41.2% - 44. 7% on the focal numbers. The arc of focal hyperplasia （ mm^2/cm^2 ） less than model group, P 〈 0. 05 ; the inhibitive rate of focal hyperplasia was 52. 72% - 60. 04%. We observed that Huqisan reduced cells in DNA synthesis phase, which percentage descend, P 〈0. 05. However, the cells obviously increased in DNA pre-synthesize and replication of DNA. Condusion： This study indicated that complete prescription Huqisan inhibited obviously rat hepatic precancerous, which were aroused by DEN, and disassemble prescription hasn＇t function.