醋氨酚所致精密肝切片损伤模型的建立及CYP2E1的调节作用

中国临床药理学与治疗学 ›› 2008, Vol. 13 ›› Issue (1): 46-50.

醋氨酚所致精密肝切片损伤模型的建立及CYP2E1的调节作用

李婧婷, 汪晖, 潘晓靓, 鄢友娥, 郭喻, 张本坚

武汉大学基础医学院药理学系, 武汉 430071

收稿日期:2007-09-11 修回日期:2007-10-26 出版日期:2008-01-26 发布日期:2020-10-13
通讯作者: 汪晖, 女, 教授, 博士生导师, 从事药物代谢与毒理研究。Tel:027-68758665 　E-mail:clbwhcbd@yahoo.com
作者简介:李婧婷, 女, 硕士生, 从事肝纤维化机制研究。Tel:15810221790 E-mail:deelee221@163.com
基金资助:
国家自然科学基金资助项目(30371666)

Establishment of an acetaminophen-induced hepatotoxicity model of precision-cut liver slices and the regulation of cytochrome P450 2E1

LI Jing-ting, WANG Hui, PAN Xiao-liang, YAN You-e, GUO Yu, ZHANG Ben-jian

Department of Pharmacology, Basic Medical School of Wuhan University, Wuhan 430071, Hubei, China

Received:2007-09-11 Revised:2007-10-26 Online:2008-01-26 Published:2020-10-13

摘要/Abstract

摘要： 目的利用精密肝切片(PCLS) 技术, 建立醋氨酚所致肝切片损伤的体外模型, 并探讨CYP2E1 活性变化对肝损伤的调节作用, 为研究和筛选肝保护药物提供实验依据。方法利用振荡切片机制备PCLS, 建立培养系统。将终浓度为500 μmol/L 的醋氨酚分别与切片共孵育0 、2 、4 、6 h, 测定培养基和切片中的谷胱甘肽S-转移酶(GST) 和乳酸脱氢酶(LDH) 活性。在体给予不同剂量(0.25 、0.5 、1.0 g/kg) 的CYP2E1 诱导剂乙醇,每天一次, 连续3 d。末次给药8 h 后处死动物,取其肝脏制备PCLS, 再与500 μmol/L 醋氨酚共同孵育6 h, 检测ALT 和LDH 漏出率。正常PCLS 与不同浓度的二乙基二硫代氨基甲酸酯盐(DDTC,CYP2E1 抑制剂) (5 、10 、20 μmol/L) 、500 μmol/L 醋氨酚共同孵育6 h, 检测ALT 和LDH 漏出率。结果与常规培养组比较, 醋氨酚孵育4 、6 h 组的GST 和LDH 漏出率显著升高(P<0.01)。与醋氨酚模型对照组比较, 乙醇中、大剂量组LDH 漏出率和小、大剂量组ALT 漏出率皆升高(P<0.01,P<0.05) ;DDTC 各浓度组ALT 漏出率则降低(P<0.05)。结论 PCLS 与500 μmol/L 醋氨酚共孵育4 h 即能成功地建立体外肝切片损伤的模型。抑制CYP2E1 活性可能对醋氨酚所致的肝损伤有保护作用, 而上调CYP2E1 活性则可能加重醋氨酚所致的肝损伤。

关键词: CYP2E1, 乙醇, 醋氨酚, 精密肝切片技术, 肝损伤

Abstract: AIM: To establish an acetaminohpheninduced hepatotoxicity model using precision-cut liver slice (PCLS) technique and observe the regulation of cytochrome P4502E1 (CYP2E1) so as to provide experimental basis on investigating and screening new potential hepatoprotective drugs. METHODS: (1) The PCLS was prepared by the vibratome and co-incubated with 500 μmol/L acetaminophen for 0, 2, 4 and 6 h. The activities of glutathione S-transferase (GST) and lactic acid dehydrogenase (LDH) were measured in the medium and slices. (2) The rats were intragastrically administrated with ethanol in different dosages (0.25, 0.5, 1.0 g/kg) once a day for three days consecutively, then they were at sacrificed 8 h after the last administration and the slices were prepared. And the slices were co-incubated with 500 μmol/L acetaminophen for 6 h and then the activities of ALT and LDH were detected in the slices and medium. (3) Replace the medium after 1 h pre-incubation, the slices were co-incubated with 500 μmol/L acetaminophen and diethyldithiocarbamate(DDTC) (5, 10, 20 μmol/L) for 6 h and the activities of ALT and LDH in the slices and medium were assayed. RESULTS: Compared with regular culture group, the leakage rates of GST and LDH in acetaminophen co-incubated for 4 and 6 h groups were significantly increased (P<0.01). Compared with those in acetaminophen model group, the leakage rates of LDH of ethanol at the dosages of 0.5, 1.0 g/kg and the leakage rates of ALT of ethanol at the dosages of 0.25, 1.0 g/kg increased significantly (P<0.01, P<0.05). Moreover, the leakage rates of ALT in all concentrations of DDTC decreased remarkably (P<0.05). CONCLUSION: Co-incubation PCLS with 500 μmol/L acetaminophen for more than 4 h is a successful way to establish hepatotoxicity in vitro model. The reduction of CYP2E1 activity might protect the slices from acetaminophen-induced hepatotoxicity, while the increase of CYP2E1 activity might aggravate it.

Key words: CYP2E1, ethanol, acetaminophen, precision-cut liver slice technique, liver injury

中图分类号:

R966

李婧婷, 汪晖, 潘晓靓, 鄢友娥, 郭喻, 张本坚. 醋氨酚所致精密肝切片损伤模型的建立及CYP2E1的调节作用[J]. 中国临床药理学与治疗学, 2008, 13(1): 46-50.

LI Jing-ting, WANG Hui, PAN Xiao-liang, YAN You-e, GUO Yu, ZHANG Ben-jian. Establishment of an acetaminophen-induced hepatotoxicity model of precision-cut liver slices and the regulation of cytochrome P450 2E1[J]. Chinese Journal of Clinical Pharmacology and Therapeutics, 2008, 13(1): 46-50.

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