中药注射液体外肾细胞毒性评价的方法学探讨

目的研究柴胡注射液和丹参注射液在犬肾近端小管上皮细胞（MDCK细胞）和人胚肾细胞（HEK-293细胞）模型的毒性作用,并探讨中药注射液体外肾细胞毒性检测的实验方法。方法选用马兜铃酸A作为阳性对照,将不同厂家和批次的丹参注射液、柴胡注射液的原液及稀释液,与MDCK细胞或HEK-293细胞共同孵育,通过四甲基偶氮唑盐比色法（MTT法）量化细胞毒性,计算相对增殖率,进行中药注射液体外肾细胞毒性评价。结果在MDCK细胞模型中,两批丹参注射4个液稀释浓度下毒性分级显示为0级,一批稀释9、27、81倍后显示为0级,柴胡注射液稀释27、81倍后显示为0级;在HEK-293细胞模型中,一批丹参注射液4个稀释浓度下毒性分级显示为1级,一批稀释9、27、81倍后显示为1级,一批稀释27、81倍后显示为1级;柴胡注射液稀释27、81倍后显示为2级。结论利用肾脏细胞来源的细胞,可尝试建立中药注射液体外肾细胞毒性检测的实验方法。

Methodological evaluation of the renal cell toxicity of Chinese medicine injection

Objective To study the cytotoxicity of Salvia Injection and Bupleurum Injection in MDCK cells and HEK-293 cells and discuss the applicability of these experimental methods. Methods Aristolochic acid A was selected as a positive control drug. MDCK cells and HEK-293 cells were incubated with different concentrations of Salvia Injection or Bupleurum Injection. Then cytotoxicity was evaluated with MTT assay and relative growth rate （RGR） was calculated. Results In MDCK cell model, the RGR from two batches of Salvia Injection were shown as Class 0 after diluting 4 lev-els and one was shown as Class 0 after diluting 9, 27, 81 times. The RGR of Bupleurum Injection was shown as Class 0 after diluting 27, 81 times. Meanwhile, in HEK-293 cell model, the RGR from one batch of Salvia Injection was shown as Class 1 after diluting 4 levels. One batch of Salvia Injection was shown as Class 1 after diluting 9, 27, 81 times. One was shown as Class 1 after diluting 27, 81 times. The RGR of Bupleurum Injection was shown as Class 2 after diluting 27, 81 times. Conclusion The use of kidney-derived cells can help to establish experimental methods to test the Chi-nese medicine injection nephrotoxicity.