黄藤素对山羊子宫内膜上皮细胞的体外毒性研究

The Study on the Cytotoxicity of Palmatine on Goat Endometrial Epithelial Cells in vitro

为了探究黄藤素的体外细胞毒性,采用不同浓度的黄藤素作用山羊子宫内膜上皮细胞（EECs）后,采用MTT 法绘制细胞生长曲线；倒置显微镜下观察细胞生长状态；瑞氏—姬姆萨染色法观察细胞形态学变化并通过测定乳酸脱氢酶（LDH）含量检测细胞膜的完整性。结果显示,当黄藤素的浓度为10-227μg/mL 时,黄藤素对EECs有促增殖的作用；当浓度大于227μg/mL 时出现细胞增殖抑制现象,药物对细胞的半数增殖抑制率（IC50）为360μg/mL；瑞氏—姬姆萨染色结果显示,100、200μg/mL 的黄藤素作用于 EECs 时均未对其细胞核、细胞浆的形态产生影响；LDH 结果显示,药物浓度≥250μg/mL 时细胞 LDH 的释放量极显著高于正常细胞组（P 〈0.01）。综上所述,黄藤素作用于山羊 EECs 后呈现一定的毒性作用,且呈现显著的剂量依赖效应,作用于 EECs 的安全剂量范围为≤250μg/mL。

To explore the cytotoxicity of palmatine in vitro ,goat endometrial epithelial cells （EE-Cs）were stimulated by different concentrations of palmatine,then the growth curve of cell was drew by MTT assay which could detect the cell proliferation.The cell growth state was observed by inverted microscope,and Wright-Giemsa staining was used to observe morphological changes of the cells while LDH test was used to check the membrane permeability of EECs.The results showed that when the concentration was 10 to 227 μg/mL,palmatine had a role in promoting pro-liferation of the EECs,while when the concentration was more than 227 μg/mL,cell proliferation was inhibited,and 50% inhibitory rate of proliferation （IC50 ） of this drug on EECs was 360 μg/mL.Wright-Giemsa staining results showed that themorphology of the nucleus and cy-tolymph were not significantly changed when 100 and 200 μg/mL palmatine effected EECs.LDH results showed that when the palmatine concentration was more than 250 μg/mL,LDH content was extremely significantly higher than that of normal cell group（P 〈0.01）.In conclusion,palmatine presented certain toxic effect on EECs with a significant dose-response effect,and the safe dose range of palmatine was less than 250 μg/mL.