商陆皂苷甲致肝毒性的研究

Hepatotoxicity induced by esculentoside A

目的通过体外、体内实验相结合的方法考察商陆皂苷甲所致的肝毒性。方法 MTT法测肝细胞(L-02细胞)活力;以不同质量浓度的商陆皂苷甲与肝细胞共培养后,测细胞培养上清中的AST(谷草转氨酶)、ALP(碱性磷酸酶)和LDH(乳酸脱氢酶)水平,并在光镜下对细胞形态进行观察;Hoechst-PI双标染色观察细胞的凋亡和坏死情况;取KM种小鼠分为正常组和给药组,给药组的小鼠每天尾静脉注射10 mg/kg商陆皂苷甲,连续给药7 d,考察小鼠血清肝功能指标和肝组织的病理学变化。结果 MTT的结果显示,商陆皂苷甲能显著抑制肝细胞活力,其IC50为360.18μg/mL;400μg/mL的商陆皂苷甲能升高细胞培养上清中的AST、ALP和LDH(P<0.01)并使肝细胞出现变圆、减少和死亡的现象;300和350μg/mL的商陆皂苷甲能使肝细胞发生凋亡和坏死;体内实验表明,小鼠血清中的AST和ALT(谷丙转氨酶)水平均明显升高(P<0.01),小鼠肝脏多出现肝细胞多发灶性坏死及肝细胞再生现象。结论大剂量的商陆皂苷甲对肝脏具有一定的毒性作用。

AIM To explore the hepatotoxicity of esculentoside A from Phytolacca acinosa Roxb. in vitro and in vivo. METHODS MTT assay was used to test the hepatic cells （L-02 cell） viability. The contents of aspartate anfinotransferase （AST）, alkaline phosphatase （ALP） and lactic dehydrogenase （LDH） in hepatocytes supernatant were detected after being incubated with the different concentrations of esculentoside A and morphological changes of hepatocytes were observed by inverted phase contrast microscope. Cell apoptosis and necrosis were ob- served by Hoechst-PI staining method. After 10 mg/kg esculentoside A were given to mice for 7 days through intra- venous administration, hepatic function and histopathology in mice was investigated. RESULTS The MTr result showed that esculentoside A could obviously inhibit hepatic cells viability with the IC50 of 360. 18 μg/mL. 400 μg/mL esculentoside A could increase the levels of AST, ALP, LDH in hepatocytes supernatant （P 〈 0. 01 ） and cell morphology showed that 400 p,g/mL esculentoside A could damage the shapes of hepatocytes. Three hun- dred and 350 μg/mL esculentoside A caused cell apoptosis and necrosis. The animal experiment showed that AST and alanine aminotransferase （ALT） in blood serum increased notably （P 〈 0.01 ）. Histopathological examination revealed that most of mice＇ s livers had multiple focal necrosis of liver cells and liver regeneration. CONCLUSION Esculentoside A may cause hepatotoxicity.