均匀设计结合多元回归分析用于淫羊藿对大鼠肝毒性的影响

Hepatoxicity of Epimedii Folium in Rat Model Based on Uniform Design and Regression Analysis

目的：通过研究淫羊藿对大鼠肝脏毒性的影响,探索淫羊藿潜在肝毒性,并对造成其毒性的原因进行初步分析。方法：以基原、工艺、剂量、疗程为考察因素,采用均匀设计方法设计各实验分组,将220只SD大鼠随机分为正常组与各给药组,每组20只,雌雄各半,连续灌服对应淫羊藿提取物,灌胃过程中观察大鼠一般情况,每周定期测定体质量和摄食量2次,在各疗程给药结束后,腹主动脉取血,测定与肝功能相关的血液生化指标,大鼠处死后取肝脏称质量,计算脏脑比和脏体比,再制作肝脏石蜡切片,经苏木素-伊红（HE）染色后进行病理评分,比较各给药组与对应正常组间各指标统计学差异,并将肝脏病理评分结果作为＂金指标＂,结合其他检测指标作为参考,进行回归分析,评价各因素对肝毒性的影响。结果：与正常组相比,实验6组与实验8组大鼠直接胆红素（DBIL）,γ-谷氨酰转移酶（GGT）,总胆红素（TBIL）,肝脏质量,脏脑比值出现明显差异（P〈0.05）,肝脏病理评分较高,表现出较明显的肝损伤情况,经回归分析并综合各指标检测结果推断,朝鲜淫羊藿和巫山淫羊藿潜在肝毒性较强,工艺与肝脏毒性强度呈负相关,剂量和疗程与肝脏毒性强度呈正相关,雌性动物受肝损害较雄性动物明显;结论：朝鲜淫羊藿及巫山淫羊藿存在潜在肝毒性,并与提取工艺、给药剂量和用药疗程存在密切关系,应当在临床应用及新药研发中引起重视。

Objective： To study the potential hepatotoxicity of Epimedii Folium and explore the reasons for toxicity by the study of the hepatotoxic effect of Epimedii Folium on rats. Method： The experiment was performed by uniform design,and the study factors included original plant,process,dose and course. A total of220 SD rats were randomly divided as the normal group and the drug group,with 20 in each group,half male and female. The rats were given the corresponding drugs and observed; and on a regular basis,the weight and food intake were determined twice a week. After the administration of each course,the rats were anesthetized with 1%pentobarbital,and blood was collected from abdominal aorta and used to detect the blood biochemical indexes related to liver function. The livers were weighted,and the ratio of liver to brain and body was calculated; and then the livers were made into paraffin sections and histopathologically scored after hematoxylin-eosin（ HE） staining.The statistical differences between normal control group and drug groups were compared. With the histopathologic score as the gold indicator and other indicators for reference, the effects of each factor on the potential hepatotoxicity were evaluated by regression analysis. Result： Compared with normal group,the animals in group 6 and 8 showed significant differences in indirect bilirubin（ DBIL）,γ-glutamyltransferase（ GGT）,total bilirubin（ TBIL）,the liver weight and the ratio of liver to brain（ P 0. 05）,and had a higher liver pathological score,which indicated a severer hepatotoxicity. After the regression and comprehensive analysis,under the experiment conditions,epimedium koreanum nakai and epimedium wushanense had a stronger toxicity,the process was negatively correlated with the liver damage degrees,the dose and course were positively correlated,and females were more seriously damaged than males. Conclusion： Epimedium koreanum nakai and epimedium wushanense have a potential hepatotoxicity,which is closely correlated with the four factors. The potential hepatotoxicity of Epimedii Folium shall be given more attention in clinical application and drug development.