金钱草不同溶剂提取物对雷公藤毒性抑制作用的比较

Comparison of Detoxification Effect of Different Solvent Extracts of Lysimachiae Herba on Toxicity Induced by Tripterygii Radix et Rhizoma

目的:考察金钱草不同提取物对雷公藤主要毒性肝损伤的减毒作用,优选提取溶剂。方法:将90只雄性SPF级昆明种小鼠按体质量随机分为9组,即正常组、金钱草水提物组、金钱草30%乙醇提取物组、雷公藤组、雷公藤配伍金钱草水提取物组、雷公藤配伍金钱草30%乙醇提取物组、雷公藤配伍金钱草60%乙醇提取物组、雷公藤配伍金钱草95%乙醇提取物组、雷公藤配伍金钱草乙酸乙酯提取物组,其中雷公藤与金钱草的剂量均按生药计分别为2,1 g·kg^(-1),正常组给予等体积的溶媒,各组均采用灌胃给予,连续14 d,末次给药24 h取血和肝组织,采用酶联免疫吸附测定(ELISA)检测小鼠血清生化指标及肝脂质过氧化/抗氧化指标,同时借助主成分分析方法整体评价金钱草提取物对雷公藤肝损伤的减毒作用及其作用机制。结果:与正常组比较,雷公藤组小鼠血清丙氨酸氨基转移酶(ALT),天冬氨酸氨基转移酶(AST),碱性磷酸酶(ALP),肝丙二醛(MDA)水平,以及由以上4个指标降维产生的肝损伤综合得分(Z值)均显著升高(P<0.01),肝抗氧化指标总超氧化物歧化酶(T-SOD),谷胱甘肽-过氧化物酶(GPX),谷胱甘肽-S转移酶(GST)水平均显著降低(P<0.01);与雷公藤组比较,分别给予金钱草2种溶剂(水,30%乙醇)提取物干预后,血清ALT,AST,ALP及肝MDA水平均明显降低(P<0.05,P<0.01),肝T-SOD,GPX,GST水平显著升高(P<0.01),给予金钱草2种溶剂(60%乙醇,95%乙醇)提取物干预后,血清ALT,AST,ALP水平均显著降低(P<0.01),肝GPX水平显著升高(P<0.01),给予金钱草乙酸乙酯提取物干预后,仅能明显降低血清AST水平(P<0.05),显著升高肝GPX水平(P<0.05);而给予金钱草不同溶剂(水,30%乙醇,60%乙醇,95%乙醇,乙酸乙酯)提取物干预后均能显著降低肝损伤综合得分(P<0.01)。结论:金钱草不同溶剂(水,30%乙醇,60%乙醇,95%乙醇,乙酸乙酯)提取物均可不同程度逆转雷公藤引起的肝损伤,其中以水和30%乙醇作为溶剂时的减毒作用为优,尤以水作为提取溶剂为更佳,且随着提取溶剂乙醇含量的升高或脂溶性的增强,肝损伤呈下降趋势。

Objective:To investigate the effect of different extracts of Lysimachiae Herba on the main toxicity induced by Tripterygii Radix et Rhizoma. Method: Ninety male SPF Kunming mice were randomly divided into 9 groups according to their body weight,control group,Lysimachiae Herba water extract group,Lysimachiae Herba 30% ethanol extract group,Tripterygii Radix et Rhizoma group,Tripterygii Radix et Rhizoma combined with Lysimachiae Herba water extract group,Tripterygii Radix et Rhizoma combined with Lysimachiae Herba 30% ethanol extract group,Tripterygii Radix et Rhizoma combined with Lysimachiae Herba60% ethanol extract group,Tripterygii Radix et Rhizoma combined with Lysimachiae Herba 95% ethanol extract group and Tripterygii Radix et Rhizoma combined with Lysimachiae Herba ethyl acetate extract group. The dosage of Tripterygii Radix et Rhizoma and Lysimachiae Herba were 2,1 g·kg^(-1) based on crude drugs,respectively. The control group was given an equal volume of solvent,and each group was given by gavage for14 consecutive days. The blood and liver tissues were taken 24 hours after the last administration. The enzyme linked immunosorbent assay(ELISA)was used to detect serum biochemical indexes and liver lipid peroxidation/antioxidant indexes in mice. Meanwhile,principal component analysis was used to evaluate the attenuating effect and the mechanism of Lysimachiae Herba extract on toxicity of Tripterygii Radix et Rhizoma. Result:Compared with control group,Tripterygii Radix et Rhizoma caused the levels of alanine aminotransferase(ALT),aspartic acid amino transferase(AST),alkaline phosphatase(ALP)in serum of mice,and the levels of malondialdehyde(MDA)in liver,and comprehensive score of toxicity(Z value)produced by the above four indexes increased significantly(P<0.01). The levels of total superoxide dismutase(T-SOD),glutathioneperoxidase(GPX),glutathione-S transferase(GST)decreased significantly(P<0.01)in liver. Compared with Tripterygii Radix et Rhizoma group,after intervention with extracts of two solvents(water,30% ethanol)of Lysimachiae Herba,the levels of serum ALT,AST,ALP and liver MDA were significantly decreased(P<0.05,P<0.01), while the levels of liver T-SOD, GPX and GST were significantly increased(P<0.01). After intervention with extracts of two solvents(60% ethanol,95% ethanol)of Lysimachiae Herba,the levels of serum ALT,AST,ALP were significantly decreased(P<0.01), and liver GPX levels were significantly increased(P<0.01). After the intervention with ethyl acetate extract of Lysimachiae Herba,only the level of serum AST was significantly decreased(P<0.05)and the level of GPX was significantly increased(P<0.05).After the intervention with extracts of different solvents(water,30% ethanol,60% ethanol,95% ethanol,ethyl acetate)of Lysimachiae Herba,it can significantly reduce the comprehensive score of toxicity(P<0.01). The overall decline rates of toxicity were 127.5%,113.4%,98.1%,56.3% and 31.0% respectively. Among them,the toxicity reduction rate of the extracts with water as a solvent was 14.1%,29.4%,71.2%,96.5% higher than those of other solvent extracts with ethanol. Conclusion: The extracts of different solvents(water,30%ethanol,60% ethanol,95% ethanol and ethyl acetate)of Lysimachiae Herba can reverse the toxicity induced by Tripterygii Radix et Rhizoma in varying degrees. Among them,water and 30% ethanol are the best solvents for detoxification, especially water as the extraction solvent, and with the increase of ethanol content or fat solubility of extraction solvent,the detoxification shows a downward trend.