大鼠肝纤维化精密切片技术的建立

中国临床药理学与治疗学 ›› 2004, Vol. 9 ›› Issue (8): 911-913.

大鼠肝纤维化精密切片技术的建立

郭喻, 汪晖

武汉大学医学院药理学系, 武汉430071, 湖北

收稿日期:2004-06-09 修回日期:2004-07-13 出版日期:2004-08-26 发布日期:2020-11-20
通讯作者: 汪晖,女,教授,博士生导师,研究方向:生化药理和药物代谢。Tel:027-87331875 E-mail:clbwhcbd@yahoo.com
作者简介:郭喻,女,硕士生,研究方向:生化药理和药物代谢。
基金资助:
湖北省自然科学基金资助项目(№2001ABB176)

Establishment of precision-cut fibrotic liver slice technique

GUO Yu, WANG Hui

Department of Pharmacology, Medical College of Wuhan University, Wuhan 430071, Hubei, China

Received:2004-06-09 Revised:2004-07-13 Online:2004-08-26 Published:2020-11-20

摘要/Abstract

摘要： 目的: 建立肝纤维化精密切片技术,摸索最佳体外培养条件,用于体外肝脏异物代谢和药物相互作用研究。方法: 建立大鼠复合因素(高脂、酒精和CCl₄)肝纤维化模型,制备肝纤维化状态下的肝切片,建立培养系统,以培养基中乳酸脱氢酶(LDH)漏出量、肝组织中谷胱甘肽S-转移酶(GST)活性、肝细胞噻唑蓝(MTT)还原能力为指标,观察切片厚度、培养液pH 值和培养时间对肝切片活力的影响。结果: 大鼠经复合因素处理后,3 周即可造成肝纤维化早期病理改变。在切片厚度300 μm、培养液pH 7.0的条件下,肝切片在6 h 内能够维持最低的LDH 漏出量和最高的GST 活性、MTT 还原量。结论: 肝纤维化状态下的精密肝切片技术,其切片厚度300 μm、培养液pH 7.0、培养时间6 h 时为最佳培养条件。

关键词: 肝纤维化, 精密肝切片, 大鼠, 培养系统, 活力

Abstract: AIM: To establish the technique of precision-cut fibrotic liver slice (PCLS) and grope the optimal cultural conditions for researching the liver xenobiotic metabolism in vitro and the drug interaction.METHODS: Complex factors (higher fat diet, alcohol and CCl4) were used to make the animal model of liver fibrosis.Fibrotic liver slices were prepared and cultivation system was established. Lactate dehydrogenase (LDH) leakage, glutathione S-transferase (GST) activity and 3[ 4, 5-Dimethythiazole-2-yl]-2, 5-diphenyltetrazolium bromide (MTT) reduction were chosen as indexes to assess the viability of the slice in different thickness, medium pH and cultural time.RESULTS: Rats were in earlier hepatic fibrosis after administration for 3 weeks.When the thickness of slices was 300 μm and medium pH was 7.0, the LDH leakage, GST activity and MTT reduction could maintain on a steady level in 6 h.CONCLUSION: A 300 μm of thickness, 7.0 of medium pH and 6 h of cultural time are the optimal slicing and culturing conditions for fibrotic liver slice.

Key words: hepatic fibrosis, precision-cut liver slice, rat, incubation system, viability

中图分类号:

R965.2

郭喻, 汪晖. 大鼠肝纤维化精密切片技术的建立[J]. 中国临床药理学与治疗学, 2004, 9(8): 911-913.

GUO Yu, WANG Hui. Establishment of precision-cut fibrotic liver slice technique[J]. Chinese Journal of Clinical Pharmacology and Therapeutics, 2004, 9(8): 911-913.

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