精密肝切片中乙醛活化肝星状细胞模型的建立

中国临床药理学与治疗学 ›› 2005, Vol. 10 ›› Issue (8): 913-916.

精密肝切片中乙醛活化肝星状细胞模型的建立

武晓茜, 汪晖, 郭喻, 廖长秀, 吴勇

武汉大学基础医学院药理学系, 武汉 430071, 湖北

收稿日期:2005-06-20 修回日期:2005-07-22 发布日期:2020-11-22
通讯作者: 汪晖,女,教授,博士生导师,研究方向:生化药理与药物代谢。Tel:027-87331875 E-mail:clbwhcbd@yahoo.com
作者简介:武晓茜,女,硕士研究生,研究方向:生化药理与药物代谢。Tel:027-68758665 E-mail:ciciail@163.com
基金资助:
国家自然科学基金项目(No30371666)和湖北省自然科学基金资助项目(No2001ABB176)

Establishment of hepatic stellate cell activated model by acetaldehyde in precision-cut liver slices

WU Xiao-qian, WANG Hui, GUO Yu, LIAO Zhang-xiu, WU Yong

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

Received:2005-06-20 Revised:2005-07-22 Published:2020-11-22

摘要/Abstract

摘要： 目的: 在精密肝切片中利用乙醛激活肝星状细胞,建立一种星状细胞激活的体外模型,为研究和筛选阻抑肝纤维化发生的药物奠定基础。方法: 利用振荡切片机制备精密肝切片,建立培养系统。以700μmol·L^-1乙醛孵育切片,培养0、2、4、6h,测定培养基和组织匀浆中谷胱甘肽S-转移酶(GST)、乳酸脱氢酶(LDH)和羟脯氨酸(Hyp)含量,观察病理切片中α-平滑肌肌动蛋白(α-SMA)表达。结果: 与0h相比,乙醛培养2、4、6h培养基的GST、LDH漏出量均显著升高(P<0.05)。切片组织Hyp含量6h较0h相比明显升高(P<0.05)。免疫组化片镜下可见4、6h有α-SMA阳性细胞表达,图像分析显示面积和阳性率较0h明显增加(P<0.05)。结论: 精密肝切片与终浓度为700μmol·L^-1乙醛共孵育6h可活化切片中星状细胞,该技术可作为良好的体外肝星状细胞激活模型。α-SMA免疫组化是一鉴定体外星状细胞激活的可靠指标。

关键词: 精密肝切片, 乙醛, 肝星状细胞, 激活, 平滑肌肌动蛋白

Abstract: AIM: To activate hepatic stellate cells (HSC) in vitro in precision-cut liver slices (PCLS) stimulated by acetaldehyde for studying and screening anti-fibrotic drugs.METHODS: PCLS were prepared by the vibratome, and incubated with 700 μm·L^-1 acetaldehyde for 0, 2, 4 and 6 h.The medium and homogenate were retained to determine glutathione S-transferase (GST) activity, lactate dehydrogenase (LDH) leakage and hydroxyproline (Hyp) content.PCLS were prepared for paraffin sections.The expression of α-smooth muscle actin (α-SMA) was evaluated by immunohistochemistry, and the result was analyzed with image analysis software.RESULTS: The leakages of GST and LDH were increased significantly compared with those in 0 h group (P < 0.05).Exposure to the acetaldehyde for 6 h, the content of Hyp in tissue slices was 1.5-fold of those in 0 h (P < 0.05).The immunohistochemical expression of α-SMA positive cells were observed in 4 and 6 h groups by the light microscope.Image analysis showed that there were significant differences in the areas and positive rates (P <0.01).CONCLUSION: Incubated with 700 μm·L^-1 acetaldehyde during 6 h, HSC is activated in PCLS.The immunohistochemical technology of α-SMA is a reliable method of identifying activated HSC in PCLS.

Key words: precision-cut liver slice, acetaldehyde, hepatic stellate cell, activate, α-SMA

中图分类号:

R966

武晓茜, 汪晖, 郭喻, 廖长秀, 吴勇. 精密肝切片中乙醛活化肝星状细胞模型的建立[J]. 中国临床药理学与治疗学, 2005, 10(8): 913-916.

WU Xiao-qian, WANG Hui, GUO Yu, LIAO Zhang-xiu, WU Yong. Establishment of hepatic stellate cell activated model by acetaldehyde in precision-cut liver slices[J]. Chinese Journal of Clinical Pharmacology and Therapeutics, 2005, 10(8): 913-916.

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