京尼平苷和京尼平的肝毒性比较研究

Comparative study on hepatotoxicity of Geniposide and Genipin

目的比较常规剂量下不同给药周期京尼平苷和京尼平的肝毒性。方法小鼠实验:将120只昆明种雄性小鼠按体重随机分空白组、对照组及低、中、高剂量实验组。空白组灌胃给予10 mL·kg^(-1)羧甲基纤维素钠,对照组灌胃给予50 mg·kg^(-1)京尼平苷,低、中、高剂量实验组分别灌胃给予25,50和100 mg·kg^(-1)京尼平。每组8只,每天给药1次,分别干预1,2和4周。给药结束后,检测肝功能相关血清生化指标,观察肝组织病理学变化。细胞实验:用含0.1,1.0,10.0,100.0和1 000.0μg·mL^(-1)京尼平苷及京尼平的培养基分别干预人源性HepaRG肝细胞24 h,通过噻唑蓝比色法及测定乳酸脱氢酶(LDH)的释放量评估细胞活力。结果与空白组相比,给药1周,低剂量实验组小鼠血清GPT和GOT水平有所升高;随着给药剂量增大及给药时间的延长,京尼平致使血清GPT、GOT和ALP水平均显著升高(均P<0.05)。肝病理结果显示:给药2周,实验组小鼠出现肝细胞肿胀、炎性细胞浸润及肝细胞坏死等明显的病理学改变;给药4周,小鼠肝病理进一步加重。与京尼平所致肝毒性相比,京尼平苷仅在给药2和4周时,使GOT水平显著升高,GPT和ALP虽有所升高,但差异均无统计学意义(均P>0.05)。给药4周,肝组织才产生炎性细胞浸润。在细胞实验中,1μg·mL^(-1)京尼平呈现出明显的肝毒性,而京尼平苷≥100μg·mL^(-1)才表现出明显的细胞毒性,且其所致毒性明显低于相同浓度下京尼平所致肝细胞毒性。结论常规剂量下京尼平是栀子主要致肝毒性的物质,京尼平苷的肝毒性可能与其水解为京尼平有关。

Objective To compare the hepatotoxicity of Geniposide and Genipin in different administration cycles at conventional doses.Methods Mouse experiment:A total of 120 male Kunming mice were randomly divided into blank group,control group and experimental-L,-M,-H groups.The blank group was gavaged with 10 m L·kg^(-1) sodium carboxymethyl cellulose,the control group was gavaged with 50mg·kg^(-1) Geniposide and the experimental-L,-M,-H groups were gavaged with 25,50 and 100 mg·kg^(-1) Genipin.Eight mice in each group were given medicine once a day for 1,2 and 4 weeks,respectively.After administration,serum biochemical indexes related to liver function were detected and histopathological changes of liver were observed.Cell experiment:Human HepaRG cells were respectively interfered with different concentrations (0.1,1.0,10.0,100.0 and1 000.0μg·mL^(-1)) of Geniposide and Genipin medium for 24 h,cell viability were evaluated by thiazolium blue colorimetric and lactate dehydrogenase (LDH) release.Results Compared with the blank group,the experimental-L group can increase the serum glutamic-pyruvic transaminase (GPT) and glutamic-oxaloacetic transaminase (GOT) levels in mice after 1 week;GPT,GOT and alkaline phosphatase (ALP)were significantly increased with increasing dose and prolonging time (all P<0.05).The liver pathology showed that the experimental groups mice had obvious pathological changes such as hepatocyte swelling,inflammatory cell infiltration and hepatocyte necrosis after 2 weeks.After 4 weeks of administration,mice’liver pathology was further aggravated.Compared with the hepatotoxicity caused by Genipin,only the level of GOT significantly increased after Geniposide administration for 2 and 4 weeks.Although the levels of GPT and ALP increased,there was no statistical difference (all P>0.05).After 4 weeks of administration,inflammatory cell infiltration occurred in liver tissue.Similarly,in the cell experiment,1μg·mL^(-1) Genipin showed obvious hepatotoxicity,while Geniposide≥100μg·mL^(-1).The toxicity was significantly lower than that of Genipin at the same concentration.Conclusion Genipin is the main hepatotoxic substance.The hepatotoxicity of Geniposide may be related to its hydrolysis to Genipin.