提取溶剂对何首乌肝细胞毒性的影响

Influence of Extracting Solvent on Hepatocytes Toxicity of Polygonum multiflorum

摘要:

目的: 比较不同溶剂提取何首乌对肝细胞毒性的差异,探讨提取方式对何首乌安全性的影响。方法: 制备不同提取方式的何首乌样品,均按生药量的5 g·L^-1给药,以CCK-8法检测人肝细胞(L02细胞)抑制率为评价指标,考察提取溶剂,溶剂量和提取时间3个因素对何首乌肝细胞毒性的影响。同时建立何首乌的UPLC指纹图谱,采用偏最小二乘法(PLS)回归分析肝细胞毒性与化学成分之间的关系。结果: 建立了基于CCK-8比色的何首乌对人正常肝细胞毒性的检测方法。何首乌75%乙醇提取物的肝细胞毒性显著＞水提取(P<0.01)。进一步正交试验显示,乙醇浓度对何首乌肝细胞毒性具有显著性影响(P<0.01),提取物肝细胞毒性顺序:50%乙醇提取物>75%乙醇提取物>95%乙醇提取物。通过偏最小二乘法回归分析,发现了2个与毒性密切相关的色谱峰。结论: 不同提取溶剂对何首乌肝细胞毒性有较大影响,50%乙醇提取物毒性最强,而何首乌泡酒主要采用50度左右的白酒,提示何首乌泡酒服用的方式可能有较大肝损害风险,应予以高度重视。

Abstract:

Objective: The difference of extracting solvent to hepatocytes toxicity of Polygonum multiflorum was researched to probe the influence of different usages to the safety of P. multiflorum. Method: The inhibition rate of P. multiflorum samples to human hepatocytes (L02 cell line) with different extracting methods was detected by CCK-8 method. The dose of all the samples was 5 g·L^-1 counted on the quantity of crude material. The influence of extracting solvent, the amount of solvent and extracting duration on hepatocytes toxicity was investigated by orthogonal test. UPLC fingerprints of the P. multiflorum samples were acquired and then analyzed with relationship to hepatocytes toxicity by partial least squares (PLS) analysis. Result: The method to detect the hepatocytes toxicity of P. multiflorum was established based on the CCK-8 colorimetry. The hepatocytes toxicity of the sample extracted with 75% ethanol was much greater than that of the sample extracted with water. The concentration of ethanol investigated in orthogonal test revealed significant influence to the hepatocytes toxicity, and the toxicity sequence of the samples was:50% ethanol>75% ethanol>95% ethanol. According PLS analysis there are two contents relative to hepatocytes toxicity. Conclusion: The extracting solvent showed significant influence on the hepatocytes toxicity of P. multiflorum and the sample extracted with 50% ethanol was of much toxic potential. It should be cautious about the high risk of liver damage of P. multiflorum, considering people usually used the herb by soaking with wine around 50 degree.