决明子的不同提取物及炮制品对四氯化碳致小鼠急性肝损伤的保护作

【摘要】   目的观察不同极性溶剂的系列提取物、总蒽醌中的游离型和结合型蒽醌蒽醌苷)及不同的炮制品的保肝作用。方法将KM小鼠随机分组,分成正常组、模型组、联苯双酯组(0.1 g/kg)、各决明子组(11 g生药/kg),连续给药9 d,末次给药后约10 h,除正常组外,其它各组均腹腔注射0.15%四氯化碳溶液10 ml/kg。16 h后采用比色法测定血清中转氨酶ALT,AST的含量。结果以不同极性溶剂提取的5种决明子提取物中,除石油醚部分外,其余各组均有显著降低转氨酶活性作用;游离型和结合型蒽醌均有显著的降低转氨酶活性作用决明子的生、炒两类炮制品也同样有显著的降酶作用,但生品略优。结论 总蒽醌类成分是保肝降酶的主要活性成分;除总蒽醌外尚有其他多种保肝降酶的活性成分,有待于对不同提取物作进一步的活性成分追踪研究;决明子的生、炒两类炮制品在用于保肝降酶活性时以生品为佳。

【关键词】 决明子 决明子提取物 蒽醌类 炒决明子 保肝作用

Abstract:ObjectiveTo investigate the protective effects of different extracts and processed products of Semen Cassiae on the acute liver injury induced by CCl4. MethodsKM mice were divided into normal group, control group, biphenyl dimethylester group (0.1 g/kg) and Semen Cassiae group (11g crude drug/kg), which were processed by intragastric administration for 9 days. 0.15%CCl4 oil solution (10 ml/kg, diluted with plant oil) was injected into the mice of all groups but normal group by intraperitoneal injection 10hr after last administration. The contents of ALT and AST in the serum were determined by chromatometry 16hr after administration of CCl4. ResultsThe five kinds of extracts extracted by different heteropolarity dissolvents but ligarine extract, the free and conjugated anthraquinones significantly decreased the activities of aminopherase. The processed products of Semen Cassiae, the crude and the parched, significantly decreased the activities of aminopherase too. The effect of the crude on the decrease of aminopherase activities was prior to the parched slightly. ConclusionAll of the anthraquinones in Semen Cassiae was the primary active component at the protective effects on liver and decrease of the aminopherase activities and there would be other active components besides all anthraquinones. The follow—up of active components in other extracts but ligarine extract would be urgent at protective effects on liver in the future. The crude may be optimal between two processed products of semen cassiae if Semen Cassiae is used for the protective aim on liver and decrease of aminopherase activities.   Key words:Semen cassiae; Extract of semen cassiae; Anthraquinones; Parched semen cassiae; Protective effect on liver   决明子为豆科草本植物决明或小决明的干燥成熟的种子,为常用中药材,其性味甘、苦、咸、微寒,归肝、肾、大肠经。具有清肝明目和润肠通便的功效[1]。现代研究认为,决明子中的化学成分主要有蒽醌类、多糖类、萘并吡酮类、氨基酸类等[2,3],药理作用主要有降血脂、降血压、润肠通便等[3,4],在保肝方面主要对总蒽醌类、多糖类、醇提物、水提物进行了研究[5~7],但对不同极性溶剂的系列提取物、总蒽醌中的游离型和结合型蒽醌蒽醌苷)及不同的炮制品的保肝作用却未见报道,本文就上述目前文献未及内容,以CCl4肝损伤为模型进行了保肝作用研究,为丰富和完善决明子保肝作用提供科学的理论依据。

1 器材。

1.1 药品与试剂   决明子药材购自江苏江浦,经鉴定为豆科植物决明Cassia Obtusifolia L.的干燥成熟种子。

联苯双酯由浙药集团新昌制药厂生产,临用前用蒸馏水于研钵中研磨制成5 mg/ml的混悬液。

丙氨酸氨基转移酶(ALT)、天冬氨酸氨基转移酶(AST)试剂盒购自南京建成生物科技有限公司;四氯化碳石油醚、二氯甲烷、正丁醇、乙醇、氯仿均为分析纯(市购);食用油(市购)。

1.2 动物KM小鼠,体重18~22 g,雄性,由南京中医药大学实验动物中心提供。

1.3 仪器2401型紫外可见分光光度计(SHIMADZU)。

2 方法与结果。

2.1 决明子各提取部位四氯化碳致急性肝损伤的保护作用

2.1.1 决明子各提取部位的分离对决明子饮片进行系统溶剂分离,分为四个部位,即石油醚部位、二氯甲烷部位正丁醇部位、乙醇部位。均用淀粉配成混悬液,备用。

2.1.2 决明子水煎液的制备取生决明子粉(过20目筛)27.5 g,以8倍量水煎煮两次,离心,过滤,滤液减压浓缩至50 ml。

取KM小鼠70只,体重18~22 g,雄性,随机分为7组,分别为正常组;联苯双酯组(0.1 g/kg);四氯化碳模型组(11g生药/kg);石油醚提取物组(11 g生药/kg);二氯甲烷提取物组(11 g生药/kg);正丁醇提取物组(11 g生药/kg);乙醇提取物组(11 g生药/kg);水煎液组(11 g生药/kg)。各组每天每鼠按0.2 ml/10 g灌胃给药1次,正常组和四氯化碳模型组给予等体积生理盐水,连续9 d。末次给药后约10 h,除正常组外,其它各组均腹腔注射0.15%四氯化碳油溶液10 ml/kg,同时禁食、自由饮水。16 h后由眼眶静脉丛取血,分离血清,按试剂盒说明书采用比色法测定血清中ALT,AST的含量。结果见表1。表1 决明子各提取部位四氯化碳致急性肝损伤的保护作用(略)。

结果表明,决明子的二氯甲烷提取物正丁醇提取物、乙醇提取物及水煎液能显著降低小鼠血清中的升高的ALT值;正丁醇提取物、乙醇提取物及水煎液能显著降低小鼠血清中升高的AST值,其中正丁醇部位四氯化碳致急性肝损伤的保护作用最为显著。

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