蛇床子素对小鼠实验性肝损伤的干预效应

背景蛇床子素是一种广泛存在于伞形科植物中的香豆素,本课题组曾报道蛇床子素具有Ca2+拮抗作用和抗炎、抗氧化等作用,另据报道蛇床子素能抑制肝肿瘤等所致血清黄嘌呤氧化酶等升高.目的观察蛇床子素对四氯化碳诱发的小鼠肝损伤模型肝损伤的影响.设计完全随机对照动物实验.单位赣南医学院药理教研室,赣南师范学院体育系.材料选用昆明种小鼠40只,雌雄不拘,体质量为(20±2)g.实验药物蛇床子素为成都龙泉高科天然药业有限公司提供.方法实验于2005-03/07在赣南医学院药理教研室完成.实验小鼠以随机数字表法分成4组,即对照组,四氯化碳模型组,蛇床子素50,100 mg/kg剂量组,每组10只.对照组和模型组分别腹腔注射生理盐水10mL/kg;蛇床子素50mg/kg剂量组小鼠腹腔注射蛇床子素50mg/kg;蛇床子素100 mg/kg剂量组小鼠腹腔注射蛇床子素100 mg/kg.1次/d,连续15 d.末次给药后2 h,正常组腹腔注射容量花生油,其余各组均腹腔注射1 g/L四氯化碳花生油溶液10 mL/kg1次.禁食,自由饮水,16 h后各组小鼠麻醉下断头取血.测定血清谷丙转氨酶、谷草转氨酶活性、肝组织丙二醛含量,并观察肝组织病理变化.主要观察指标各组小鼠血清中谷丙转氨酶、谷草转氨酶活性、肝组织丙二醛含量,并观察肝组织病理变化.结果40只小鼠均进入最后结果分析.①给四氯化碳诱导16 h后,模型组小鼠血清中谷丙转氨酶和谷草转氨酶均高于对照组(P＜0.001).蛇床子素组呈剂量依赖性地抑制四氯化碳所致的谷丙转氨酶的升高,100 mg/kg剂量组显著性抑制四氯化碳所致的谷丙转氨酶的升高(P＜0.05),蛇床子素50,100 mg/kg剂量组均可显著性抑制四氯化碳所致的谷草转氨酶的升高(P＜0.01～0.001).②模型组丙二醛含量的增加(P＜0.05),蛇床子素100 mg/kg剂量组的丙二醛的含量接近对照组,可明显降低四氯化碳肝损伤小鼠肝脏丙二醛含量(P＜0.05),蛇床子素50 mg/kg组虽没有显著性抑制四氯化碳所致的丙二醛含量的升高,但有抑制四氯化碳所致肝损伤丙二醛含量的趋势.③四氯化碳肝损伤模型组肝组织明显损伤,肝细胞浊肿、气球样变性,病变以肝小叶中央静脉周围为重.肝小叶内可见轻度、中度肝细胞点状、碎片状坏死;坏死区内有中、重度淋巴细胞和浆细胞浸润,汇管区内有单核细胞浸润.蛇床子素50,100 mg/kg组也有不同程度的肝细胞破坏、炎性细胞浸润等肝损伤病变,但与模型组比较损伤程度明显减轻,尤其是100 mg/kg剂量组表现更为明显.结论蛇床子素能保护四氯化碳所致小鼠肝损伤,表现为降低血清谷丙转氨酶和谷草转氨酶活力和肝脏丙二醛含量.

Interventional effect of osthol on experimental liver injury in mice

BACKGROUND: Osthol is a simple coumarin from Cnidium monnier (L.)Cusson which has been long used of in China as a herbal medicine for arthritis. We have previously observed protective effects of osthol on Ca2+antagonism, oxidative stress and inflammation. And other researches reported that it could inhibit increase of serum xanthine oxidase induced by liver tumor.OBJECTIVE: To investigate the protective effect of osthol on carbon tetrachloride (CCl4)-induced liver injury of mice.DESIGN: Completely randomized controlled study.SETTING: Department of Pharmacology, Gannan Medical College; Physical Education, Gannan Normal College.MATERIALS: A total of 40 Kunming mice were of both genders and weighing (20±2) g. Osthol was provided by Chengdu Longquan High-Tech Natural Pharmaceutical Co. Ltd.METHODS: The experiment was performed at Department of Pharmacology, Gannan Medical College from March to July 2005. Forty mice were randomly divided into control, model, osthol (ip 50 g/kg) and osthol (ip 100 g/kg) groups with 10 in each. Separately once a day for 15 consecutive days, the control and model groups were equalized injected intraperitoneally with 10 mL/kg saline and osthol groups injected intraperitoneally with 50 and 100 g/kg osthol, respectively. On 15 day just after treatment,they, except the control, were challenged with CCl4 (ip 1 g/L peanut oil solution 10 mL/kg). Then all mice were free access to water but fast food.At specified time points 16 hours after the injection of CCl4, all mice were sacrificed and blood was collected in centrifuge tubes. The serum alanine aminotransferase (ALT), aspartate aminotransferase (AST) and liver malondialdehyde (MDA) were determined, and the pathological examination of liver was observed.MAIN OUTCOME MEASURES: Contents of serum ALT, AST, MDA and pathological examination.RESULTS: Forty mice were involved in the final analysis. ① At 16 hours after CCl4 induction, contents of ALT and AST were higher in model group than those in control group (P ＜ 0.001). The increase in the contents of ALT and AST was inhibited by osthol in a dose-dependent manner, especially in 100 mg/kg osthol group (P ＜ 0.05). Both 50 mg/kg and 100 mg/kg osthol could inhibit the increase of AST induced by CCl4 (P ＜ 0.01-0.001).② Content of MDA in model group was increased (P ＜ 0.05), and content of MDA in 100 mg/kg osthol group was similar to that in control group.100 mg/kg osthol could decrease content of MDA, 50 mg/kg osthol could increased the content; however, it still had the tendency of decrease. ③Effect of osthol on histopahtological changes, the livers of CCl4intoxicated mice showed massive fatty change, gross necrosis, broad infiltration of the lymphocytes, and Kupffer cells around the central vein, loss of cellular boundary. The histological pattern of the livers of the mice treated with 100 mg/kg and 50 mg/kg osthol showed a mild degree of fatty change,necrosis and lymphocyte infiltration. In contrast, the inhibitory potency of 100 mg/kg osthol on the histological changes significantly higher than those models.CONCLUSION: Osthol can protect against CCl4-induced hepatotoxicity in mice through decreasing activities ALT and AST and contents of MDA.