百里香酚对山羊子宫内膜上皮细胞的体外毒活性研究

Study on the cytotoxic activity ofthymol of goat endometrial epithelial cells in vitro

关键词:  百里香酚  子宫内膜上皮细胞  细胞毒性  膜通透性

Key words:  thymol  endometrial epithelial cells  cytotoxicity  membrane permeability

摘要:

为了探究百里香酚的体外细胞毒活性。采用不同浓度的百里香酚作用山羊子宫内膜上皮细胞(endometrial epithelial cells,EECs)后,采用MT T 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-吉姆萨染色法观察细胞形态学变化；LDH法检测细胞膜的完整性。结果表明,百里香酚对EECs的毒性作用与其浓度呈剂量依赖效应(y = -0.008x 1.2202,R2 = 0.9767),随着药物浓度的增大细胞的增殖抑制率增大,药物对细胞的半数增殖抑制率(IC50)为100 μg/ mL。瑞氏-姬姆萨染色结果显示50 μg/mL、100 μg/mL的百里香酚作用于EECs时均未对其细胞核、细胞浆的形态产生明显影响,但150 μg/mL 的百里香酚作用于EECs时,细胞的细胞膜破裂,细胞死亡；LDH结果显示,药物浓度≥100 μg/mL时细胞LDH的释放量显著高于正常细胞组。总结,百里香酚作用于山羊子宫内膜上皮细胞后呈现一定的毒性作用,且毒性作用和药物的浓度有关,作用于细胞的安全范围在0-100 μg/mL,为以后百里香酚治疗奶牛子宫内膜炎的研究提供相关的实验依据。

Abstract:

To explore the cytotoxicity of thymol in vitro. Different concentrations of thymol stimulated goat endometrial epithelial cells (EECs), then drew cell curve by MTT assay which can detect the cell proliferation; The cell growth state was observed by inverted microscope; Wright Giemsa staining was used to observe morphological changes of the cells; LDH test was used to check the integrity of the cell membrane. The results, it showed a dose-dependent effect between thymol toxicity to EECs and the concentration, palmatine has a role in promoting proliferation of the EECs(y = -0.008x 1.2202,R2 = 0.9767); With the concentration of the drug increased the inhibitory rate of cell proliferation increased.And 50％ inhibitory rate of proliferation (IC50) of this drug on EECs is 100 μg/mL; Wright Giemsa staining results showed that themorphology of the nucleus andcytolymph has no significant change when 50 μg/mLand 100 μg/mL of thymol effected EECs.But when 150 μg/mL of thymol effected EECs, the cell membrane ruptured, cell death. LDH results showed that when drug concentration was more than 100 μg/mL, LDH release was significantly higher than that of normal cel group. Draw a conclusion, thymol presented certain toxic effect on goat endometrial epithelial cell in a significant dosage dependent manner, and the safe dose range of palmatine effect on EECs is less than 100 μg/mL.