原代肝、肾细胞培养在保健食品原料毒性筛选中的应用

目的建立肝、肾毒性的体外筛选方法,并研究其在保健食品原料毒性筛选中的应用。方法以原代肝细胞、原代肾细胞、CHL和HepG2细胞为平台,用MTT法检测毒物对细胞活性的半数抑制浓度IC50,用生化学方法检测培养液中酶学指标的变化,作为毒性效应标志物;用已知毒物对该方法进行验证;用该方法对大黄、苍术、红花水提物进行毒性筛选。结果该方法能够筛选出已知毒物的肝肾毒性;在本实验条件下,未发现大黄、苍术对肝肾存在靶毒性,红花可能存在潜在的肾毒性。结论本研究建立的方法可用于保健食品原料的毒性筛选;有必要对红花进行更深入的安全性研究。

Application of in vitro Screening Test in Screening the Toxicity of Raw Materials for Health Food on Primary Hepatocyte and Nephrocyte Cell Culture

Objective To establish an in vitro screening method for hepatotoxicity and nephrotoxicity and to apply the test in screening the toxicity of raw materials for health food. Methods Primary hepatocytes, primary nephrocytes, CHL and HepG2 cells were cultured. IC₅₀ values were tested by MTT, and the enzymes leakaging from cells were measured by biochemical methods and used as markers for toxicity. The method was validated by known toxic substances and was applied in screening the toxicity of raw materials for health food, such as water-extracted Rhubarb, Rhizoma Atractylodis and Safflower. Results The method was effective for screening the hepatotoxicity and nephrotoxicity. Target toxicity was not found from Rhubarb and Rhizoma Atractylodis on hepatocytes and nephrocytes. It might have potential nephrotoxicity from Safflower water-extracts. Conclusion The in vitro screening test for hepatotoxicity and nephrotoxicity was able to be used in screening raw materials for health food. Further study on evaluating the safety of Safflower water-extracts might be needed.