论文部分内容阅读
目的:通过体外实验考察薯蓣皂苷Dioscin(Dio)对肝脏的毒性。方法:①采用MTT法检测肝细胞活力;②以不同浓度的薯蓣皂苷与人肝细胞(L-02)共培养后,检测细胞上清液中的丙氨酸转氨酶(ALT),碱性磷酸酶(ALP)和乳酸脱氢酶(LDH)含量,并采用倒置相差显微镜观察细胞形态;③采用Hoechst-PI染色法观察细胞凋亡;④KM种小鼠分为正常组和给药组,给药组小鼠每天尾静脉注射10μg/ml薯蓣皂苷,连续7d,观察血清肝功能和肝组织病理学变化。结果:①MTT法显示,薯蓣皂苷对肝细胞活力有明显的抑制作用,IC50为2.25μg/ml;②10μg/ml的薯蓣皂苷能升高肝细胞上清中的AST、ALP、LDH,2.5和5μg/ml的薯蓣皂苷能升高肝细胞上清中的ALP,随着薯蓣皂苷浓度剂量的增加,肝细胞不同程度的减少、变圆、甚至死亡;③5和10μg/ml的薯蓣皂苷能使肝细胞不同程度的出现凋亡和坏死的现象;④整体实验表明,给药组每日从尾静脉给予薯蓣皂苷10mg/kg连续7天,小鼠血清中的AST和LDH水平出现升高,仅有1只小鼠肝脏出现肝细胞广泛水肿和2只小鼠出现偶见巨核肝细胞现象。结论:大剂量的薯蓣皂苷可能对肝脏具有一定的毒性。
Objective: To investigate the toxicity of Dioscin (Dio) on liver in vitro. Methods: (1) MTT assay was used to detect the viability of hepatocytes; (2) cotyledons of different concentrations of diosgenin and human hepatocytes (L-02) were co-cultured to detect the levels of alanine aminotransferase (ALT), alkaline phosphatase (ALP) and lactate dehydrogenase (LDH), and the morphological changes of the cells were observed by inverted phase contrast microscope. ③Hoechst-PI staining was used to observe the apoptosis of the cells. ④KM mice were divided into normal group, The mice were injected intravenously with 10μg / ml diosgenin through tail vein every day for 7 days. The changes of serum liver function and pathological changes were observed. Results: ①MTT assay showed that diosgenin significantly inhibited the viability of hepatocytes with IC50 of 2.25μg / ml; ②The diosgenin of 10μg / ml increased AST, ALP, LDH, 2.5μg / ml and 5μg / ml of diosgenin can increase the ALP in the supernatant of hepatocytes, with the dosage of diosgenin increased, the degree of hepatocyte decreased, rounded, and even died; ③ 5 and 10μg / ml of diosgenin can make different hepatocytes Degree of apoptosis and necrosis; ④ The overall experiment showed that daily administration of diosgenin 10mg / kg from the tail vein to the administration group for 7 days continuously increased the levels of AST and LDH in the serum of the mice, only 1 The liver cells of mice showed extensive edema of liver cells and occasional megakaryocytic hepatocytes occurred in 2 mice. Conclusions: High-dose diosgenin may be toxic to the liver.