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目的:探讨斑蟊素致肝脏慢性损伤的可能机制。方法:用不同剂量斑蟊素持续对昆明小鼠灌胃14天,检测其肝功能、肝脏指数和肝脏病理变化,检测ATF-6、XBP1、GRP78、ATF-4、CHOP、BAX、Bcl-2、caspase3、caspase8和caspase9蛋白的表达。用不同浓度斑蟊素连续培养人肝细胞LO2系,显微观察生长,CCK-8检测活性,检测GRP78和CHOP的mRNA的表达,检测GRP78、ATF-4、CHOP、BAX、Bcl-2、caspase3、caspase8和caspase9蛋白的表达。结果:斑蟊素(≥1.0 mg/kg)能增高小鼠转氨酶和肝脏指数,肝细胞坏死及凋亡,且程度与其剂量正相关;ATF-6、XBP1、GRP78、ATF-4、CHOP、BAX、caspase3、caspase8和caspase9蛋白的表达上调,Bcl-2蛋白的表达下调。斑蟊素(≥0.392μg/ml)抑制LO2系细胞活性,增高GRP78和CHOP基因mRNA的表达,GRP78、ATF-4、CHOP、BAX、caspase3、caspase8和caspase9蛋白的表达上调,Bcl-2蛋白的表达下调。结论:斑蟊素可能主要通过内质网应激途径诱导肝细胞凋亡从而导致肝脏的慢性损伤。
Objective: To investigate the possible mechanism of cantharidin-induced chronic liver injury. Methods: Kunming mice were continuously gavaged with different doses of cantharidin for 14 days. The liver function, liver index and liver pathological changes were detected. ATF-6, XBP1, GRP78, ATF-4, CHOP, BAX and Bcl- , Caspase3, caspase8 and caspase9 protein expression. The human hepatocyte line LO2 was cultured continuously with different concentrations of cantharidin. The growth and the activity of CCK-8 were detected by microscopy. The mRNA expression of GRP78, CHOP, BAX, Bcl-2, caspase3 , Caspase8 and caspase9 protein expression. Results: Cantharidin (≥1.0 mg / kg) increased the transaminase, liver index, hepatocyte necrosis and apoptosis in mice, and the degree was positively correlated with the dose of cantharidin. ATF-6, XBP1, GRP78, ATF-4, CHOP and BAX , The expressions of caspase3, caspase8 and caspase9 were up-regulated and the expression of Bcl-2 was down-regulated. Cantharidin (≥0.392μg / ml) inhibited the activity of LO2 cells and increased the mRNA expression of GRP78 and CHOP gene. The expressions of GRP78, ATF-4, CHOP, BAX, caspase3, caspase8 and caspase9 were up- Down regulation. CONCLUSION: Cantharidin may induce hepatocyte apoptosis mainly through the endoplasmic reticulum stress pathway, leading to chronic liver damage.