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目的:观察黄药子中二萜内酯成分对大鼠肝细胞的损伤作用。方法:黄药子饮片3kg,用75%乙醇提取,氯仿萃取得萃取物22g,取萃取物18g与40ml吐温80(聚山梨酯80)研均,加蒸馏水配成0.044g/ml的混悬液。20只雄性SD大鼠分成2组:萃取物组及对照组,每组10只,萃取物组给予萃取物混悬液2.5～3ml(按体重计算,1.19g/kg)灌胃,对照组给予含同浓度吐温80的等体积蒸馏水,均2次/d,连续灌胃7d。观察大鼠外观、活动、饮食,于7d后称体重,检测谷胱甘肽过氧化物酶(GSH-PX)、过氧化氢酶(CAT)及超氧化物歧化酶(SOD),计算肝脏指数,透射电镜观察肝细胞超微结构。结果:萃取物组大鼠皮毛明显枯槁无光泽、脱毛,少动蜷卧,食量减少;萃取物组与对照组比较体重变化率分别为(0.929±0.032)%与(1.171±0.046)%,差异有统计学意义(P<0.01);2组的肝脏指数分别为14.541±2.154与8.602±0.170,差异也有统计学意义(P<0.01);2组GSH-PX、CAT及SOD分别为(157.63±57.44)U/L、(0.92±0.61)U/L和(131.12±27.54)U/L与(375.41±46.92)U/L、(6.44±2.25)U/L和(253.05±34.84)U/L,差异均有统计学意义(均P<0.01)。电子显微镜观察萃取物组大鼠可见肝细胞内脂滴和滑面内质网明显增多,线粒体肿胀、髓样改变,肝细胞表面微绒毛融合,肝血窦周围间隙结构不清。结论:黄药子萃取物中二萜内酯类成分具有肝细胞毒性,其机制可能与该成分引起线粒体的氧化损伤有关。
OBJECTIVE: To observe the effect of diterpene lactone component on the hepatocyte injury of rats in Dioscorea bulbifera. Methods: 3 kg of Huangyazi decoction pieces were extracted with 75% ethanol and extracted with chloroform to obtain 22 g of extract. 18 g of extract and 40 ml of Tween 80 (polysorbate 80) were mixed and distilled water was added to make a 0.044 g/ml suspension. Twenty male Sprague-Dawley rats were divided into two groups: the extract group and the control group, 10 in each group. The extract group was given 2.5 to 3 ml of the extract suspension (by weight, 1.19 g/kg) gavage; the control group was given An equal volume of distilled water containing the same concentration of Tween 80 was administered twice a day for 7 days. Observe the appearance, activity and diet of the rats and weigh them after 7 days. Detect glutathione peroxidase (GSH-PX), catalase (CAT) and superoxide dismutase (SOD) and calculate the liver index. Transmission electron microscopy was used to observe the ultrastructure of hepatocytes. RESULTS: The rats in the extract group were obviously dull, dull and hairless, with little movement and lying down, and their food intake decreased. The body weight change rates of the extract group and the control group were (0.929±0.032)% and (1.171±0.046)%, respectively. There was statistical significance (P<0.01); liver index in the two groups was 14.541±2.154 and 8.602±0.170, respectively, and the difference was statistically significant (P<0.01); GSH-PX, CAT and SOD in the two groups were (157.63±) 57.44) U/L, (0.92±0.61) U/L, (131.12±27.54) U/L and (375.41±46.92) U/L, (6.44±2.25) U/L, and (253.05±34.84) U/L The differences were statistically significant (all P<0.01). The electron microscopy observation showed that the lipid droplets and smooth endoplasmic reticulum in hepatocytes were significantly increased, the mitochondria were swollen, the myeloids were changed, the microvilli were fused on the surface of liver cells, and the structure around hepatic sinusoidal spaces was unclear. CONCLUSION: The diterpene lactones in the extract of Dioscorea bulbifera have hepatotoxicity. The mechanism may be related to the oxidative damage caused by this component in mitochondria.