蒙药山沉香护肝、抗氧化活性部位研究

目的研究蒙药山沉香不同提取物护肝、抗氧化作用,筛查山沉香具有护肝、抗氧化作用的活性部位。方法以浓度为95%乙醇回流提取山沉香粉末浓缩得到乙醇提取物浸膏,用水分散制成混悬液后按极性从小到大依次用石油醚、乙醚、乙酸乙酯、正丁醇按比例1∶3萃取,减压浓缩干燥后称质量用于药效学实验。将80只雄性昆明种小鼠随机分成空白组(羧甲基纤维素钠CMC-Na)、模型组(CCl4-花生油)、阳性对照组(联苯双酯)、山沉香乙醇部位提取物组、山沉香石油醚部位提取物组、山沉香乙醚部位提取物组、山沉香乙酸乙酯部位提取物组、山沉香正丁醇部位提取物组,每组10只。每天按20 mL·kg^-1的剂量灌胃给药1次,连续灌胃7 d。最后一次给药1 h后除空白组外其余组均注射0.1%CCl4-花生油(20 mL·kg^-1)造成急性肝损伤模型,空白组注射等量花生油。12 h后观察小鼠的毛色及活动情况,检测小鼠血清中谷丙转氨酶(ALT)、谷草转氨酶(AST)含量,肝匀浆中丙二醛(MDA)、谷胱甘肽过氧化物酶(GPx)、超氧化物歧化酶(SOD)含量,并观察肝组织形态外观、切片病理特征及对其进行Ishak评分。结果与空白组比较,模型组小鼠体型偏瘦,毛发黯淡无光且较稀疏,灵活度下降,容易聚堆;血清ALT、AST含量明显升高(P<0.01);肝匀浆MDA含量明显升高(P<0.01),GPx和SOD含量明显降低(P<0.01);模型组小鼠肝组织外观呈暗棕色,体积增大;病理切片表现为肝小叶中央静脉周围出现凝固性坏死、肝索肝窦排列不整齐Ishak评分明显升高(P<0.01)。与模型组比较,山沉香乙醇提取物组和乙酸乙酯提取物组小鼠毛色光泽,体型较为丰满;血清ALT、AST含量明显降低(P<0.01);山沉香乙酸乙酯提取物组小鼠肝匀浆MDA含量明显降低(P<0.01),GPx(P<0.01)和SOD(P<0.05)含量明显升高;且该组肝组织形态外观与空白组接近,病理损伤有明显改善,Ishak评分明显降低(P<0.01)。结论蒙药山沉香具有护肝作用,而且可通过抗脂质过氧化来保护肝脏,山沉香95%醇提物的乙酸乙酯提取物是其护肝、抗氧化的活性部位。

Study on the Liver Protection and Antioxidant Active Part of Syringa pinnatifolia

Objective To study liver protection and anti-oxidation effects of different extracts of Syringa pinnatifolia.Methods The Syringa pinnatifolia powder was refluxed with 95%ethanol to obtain ethanol extract.After dispersing with water,the suspension was extracted using petroleum ether,ether,ethyl acetate and n-butanol in order according to the polarities of the solvents,with a ratio of 1∶3.The obtained extracts were vacuum concentrated and weighed.Eighty male mice were randomly divided into blank group(CMC-Na),model group(CCl4-arachis oil),biphenyldiestergroup(positive control),Syringa pinnatifolia ethanol extract group,Syringa pinnatifolia petroleum ether extract group,Syringa pinnatifolia ether extract part group,Syringa pinnatifolia ethyl acetate extract group,and Syringa pinnatifolia n-butanol extract group,10 in each group.Mice were intragastrically administered once every day at a dose of 20 mL·kg^-1 for 7 days.One hour after the last administration,apart from blank group mice,the mice in other groups were injected with 0.1%CCl4-peanut oil(20 mL·kg^-1)to induce acute liver injury model.After12 hours,the coat color and activity of the mice were observed.Serum alanine aminotransferase(ALT)and aspartate transaminase(AST)levels of mice were tested;Content of malondialdehyde(MDA),activities of glutathione peroxidase(GPx)and superoxide dismutase(SOD)in the liver homogenates were detected.Liver tissue morphology and histopathological morphology were observed,and the Ishak scores were obtained.Results Compared with the blank group,the mice in the model group were thinner,the hair was dull and less dense,the elasticity was decreased,and willing to gather.The serum levels of ALT and AST in the mice were significantly increased(P<0.01).The MDA content in the liver homogenate was significantly increased(P<0.01);GPx and SOD activities were significantly decreased(P<0.01).Liver tissues were dark brown in color and increased in size.The pathological features of liver tissues of the mice in the model group showed coagulative necrosis around the central vein of the hepatic lobules,and the hepatic sinusoids were disorderly arranged,and Ishak score was significantly decreased(P<0.01).Compared with the model group,the mice in the ethanol extract and the ethyl acetate extract groups had shiny coats with fuller figures,the serum levels of ALT and AST were significantly lower(P<0.01).The MDA content in the liver homogenate of the ethyl acetate extract group was significantly lower(P<0.01);GPx(P<0.01)and SOD(P<0.05)activities were significantly increased;and the liver tissue morphology of mice in this group were close to the blank group.The pathological damage of the tissue was also significantly reduced and Ishak score was significantly increased(P<0.01).Conclusion Mongolian medicine Syringa pinnatifoliahas has liver protective and antioxidant pharmacological effects.The ethyl acetate part of the ethanol extract is the main active part.