不同来源生脉注射液体外细胞毒性的比较及原因分析

目的:比较不同来源生脉注射液体外细胞毒性的差别,寻找造成差别的主要原因。方法:将不同来源生脉注射液原液以及不同浓度的稀释液分别与L929细胞接触培养,通过倒置相差显微镜观察其形态,采用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,并进行细胞毒性评价。结果:8个厂家生产的生脉注射液原液的细胞毒性分级均为4级,为重度毒性。样品A的EC50最高,说明其细胞毒性较其他样品高。在生脉注射液所添加的辅料中,当聚氧乙烯脱水山梨醇单油酸酯(吐温80)浓度为处方量的0.5%时,其细胞毒性分级为3～4级。将吐温80稀释至0.062 5%时,其细胞毒性分级为1级,基本无细胞毒性。8个厂家生产的生脉注射液原液中吐温80的含量不一致,有些厂家的含量大于0.5%。结论:不同厂家生产的生脉注射液由于原料产地、生产条件不同等原因,其细胞毒性存在较大的差别。吐温80作为中药注射剂的辅料,在高于0.062 5%的浓度下存在细胞毒性。生脉注射液中吐温80的含量与细胞毒性可能呈一定的正相关性。

Comparison and analysis of in vitro cytotoxocity of different shengmai injections

Objective: To compare the cytotoxic response with respect to different Shengmai injections manufactured by different manufacturers and to find the main reasons that cause the differences. Method: L929 cells were cultured with various Shengmai injections which were incubated into serum-supplemented minimum essential medium at different doses. The cellular morphology was observed by phase contrast inverted microscopy and proliferation of the cells was examined using mitochondrial function methyl thiazolyl tetrazolium (MTT) assay. Relative growth rate (RGR) was calculated. Moreover, cytotoxicity was evaluated. Result: For Shengmai injections manufactured by 8 manufacturers, cytotoxicity was high, Class 4. The EC value of sample A was high than other samples. This result indicated that sample A is much more toxic than other samples. For the excipients of Shengmai injections, when the concentration of tween 80 is 0.5% of composition of drug products, cytotoxicity was classified as 3-4; while the concentration of tween 80 is 0.062 5% of composition of drug products, the cellular toxicity was classified as 1 (no cellular toxic response). The content of tween 80 of shengmai injection is different during different factories; some of it is higher than 0.5%. Conclusion: Cytotoxic results of these various injections were significantly different because of different manufacturers of drug substance and manufacturing process. As an excipient of injection, it showed cytotoxicity when the concentration is higher than 0.062 5%. The content of tween-80 and the degree of cytotoxicity in different Shengmai injections may have a positive correlation.