盐酸多巴酚丁胺注射液细胞毒性研究

Comparison of in vitro Cytotoxicity of Different Dobutamine Hydrochloride Injections

目的:比较不同来源盐酸多巴酚丁胺注射液体外细胞毒性的差别,为遴选优质药品、保障用药安全提供科学依据。方法:将不同来源国产盐酸多巴酚丁胺注射液原液以及稀释液,与L929细胞接触培养,通过倒置相差显微镜观察其形态,采用MTT法量化细胞毒性,计算相对增值率和IC50值,进行细胞毒性评价。并将国产盐酸多巴酚丁胺注射液与进口制剂进行比较。结果:8个厂家生产的盐酸多巴酚丁胺注射液原液及进口产品的细胞毒性分级均为4级。不同企业生产的盐酸多巴酚丁胺注射液的IC50值差异较大。盐酸多巴酚丁胺注射液C和F的IC50值较小,说明这两个企业产品的细胞毒性较其他样品大。进口盐酸多巴酚丁胺注射液的IC50值为99.9 mg·L-1,较其他样品高,说明其细胞毒性较其他样品低。进口盐酸多巴酚丁胺注射液中仅添加亚硫酸氢钠一种辅料,国产盐酸多巴酚丁胺注射液中添加的辅料主要为依地酸二钠、亚硫酸氢钠、盐酸半胱氨酸等。按各辅料在国产盐酸多巴酚丁酸注射液中的浓度测其细胞毒性,依地酸二钠、盐酸半胱氨酸和亚硫酸氢钠的细胞毒性分级均为3级。将依地酸二钠的浓度稀释为0.03 mg·ml-1时,其细胞毒性分级为1级,无细胞毒性。将盐酸半胱氨酸稀释到0.11 mg·ml-1时,其细胞毒性分级为1级,基本无细胞毒性。结论:不同厂家生产的盐酸多巴酚丁胺注射液由于生产条件不同、处方不同等原因,这些产品的细胞毒性存在较大的差别。与进口制剂比较,国产制剂中添加的辅料浓度较大,可适当降低其浓度,以减少药物制剂的细胞毒性,从而提高药品的安全性。

Objective：To compare the in vitro cytotoxicity of dobutamine hydrochloride injections from different factories to provide scientific evidence for the choice of high-quality medicines and the safety guarantee of drug application.Methods：L929 cells with different dobutamine hydrochloride injections at various doses were incubated together.The cellular morphology was observed under an inverted phase contrast microscope and the proliferation of the cells was quantitatively examined using MTT assay.The relative growth rate （RGR） and IC50 were calculated,and the cytotoxicity was evaluated.The difference among the injections from the domestic and abroad was compared.Results：The cytotoxicity of dobutamine hydrochloride injections produced by 8 different factories was all in grade 4,however,significant difference was shown in IC50.Sample C and F with lower IC50 showed higher cytotoxicity.IC50 of the imported injections was 99.9 mg · L-1,which was the highest among the tested injections suggesting the lowest cytotoxicity.Sodium bisulfite was the only adjuvant added in the imported injections,while EDTA-2Na,sodium bisulfite and cysteine hydrochloride were added in the domestic products.The cytotoxicity of EDTA-2Na,sodium bisulfite and cysteine hydrochloride with the same concentration in the injections was in grade 3.The cytotoxicity was grade 1 when EDTA-2Na was diluted to 0.03 mg · ml-1,and that was also grade 1 when cysteine hydrochloride was diluted to 0.11 mg · ml-1,both showing no cytotoxicity.Conclusion：The cytotoxicity of dobutamine hydrochloride injections from different factories is various.Sodium bisulfite,cysteine hydrochloride and EDTA-2Na may be relative to the toxicity of dobutamine hydrochloride injections.