南方医科大学学报 ›› 2005, Vol. 25 ›› Issue (01): 66-70.

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FK506促进异体神经匀浆激活的巨噬细胞凋亡的实验研究

杨俊1, 罗勇2, 邱小忠1, 武雷1, 余磊1, 陆云涛1, 朴英杰1, 秦建强1   

  1. 1. 南方医科大学临床解剖学研究所;
    2. 南方医科大学南方医院整形外科, 广东广州510515
  • 出版日期:2005-01-20 发布日期:2005-01-20
  • 基金资助:
    收稿日期:2004-7-26。
    基金项目:国家自然科学基金(39970833);广东省科技计划专项(2003A3020101);广东省重点实验室(2001B60107)
    作者简介:杨俊(1977- ),男,南方医科大学在读硕士研究生,电话:020-61647752,E-mail:yangjun1267@sina.com
    通讯作者:秦建强,南方医科大学解剖教研室副主任,副教授,电话:020-61689092, E-mail:jqqin@fimmu.com

FK506 promotes apoptosis of macrophages activated by homogenate of allogenic nerve

YANG Jun1, LUO Yong2, QIU Xiao-zhong1, WU Lei1, YU Lei1, LU Yun-tao1, PIAO Ying-jie1, QIN Jian-qiang1   

  1. 1. 南方医科大学临床解剖学研究所;
    2. 南方医科大学南方医院整形外科, 广东广州510515
  • Online:2005-01-20 Published:2005-01-20

摘要: 目的 研究FK506促进外周血来源的异体神经匀浆激活的巨噬细胞的凋亡。方法 取1月龄SD幼鼠,腹腔抽取法培养异体神经匀浆激活的巨噬细胞。按不同浓度的FK506分组:A组空白对照组、B组0.25ng/ml、C组0.5ng/ml、D组1.0ng/ml。将4组处理因素分别加入长有巨噬细胞的96孔板中,用MTT法检测巨噬细胞的活力情况,用倒置显微镜和荧光显微镜及透射电镜对巨噬细胞的凋亡进行形态学上的观察与鉴定。流式细胞仪检测巨噬细胞凋亡情况。结果 B组和C组均可见异体神经匀浆激活的巨噬细胞的活性减弱及细胞凋亡发生,D组可见巨噬细胞发生坏死性改变。透射电镜和荧光显微镜可观察到巨噬细胞的凋亡前期和凋亡小体出现。流式细胞仪检测B和C组凋亡率为24.6%和26.5%,均高于对照组,D组发生坏死。结论 FK506可以在早期促进异体神经匀浆激活的巨噬细胞的凋亡,从而减少或抑制周围神经异体移植后巨噬细胞所介导的免疫排斥反应。

Abstract: Objective To study the effect of FK506 in promoting apoptosis of peripheral blood-derived macrophages activated by homogenate of allogenic nerve tissues. Methods Homogenate of the allogenic nerve tissues was prepared using the sciatic nerve and injected in one-month-old SD rats, from which the macrophages activated by the homogenate were collected from the abdominal cavity and cultured in vitro. The cells were divided into 4 groups according to different concentrations of FK506 for treatment, namely 0 (group A, control group), 0.25 ng/ml (group B), 0.5 ng/ml (group C), and 1.0 ng/ml (group D). The cells of the 4 groups were inoculated into 96-well plate respectively for detecting the viability of the macrophages by MTT assay and for morphological evaluation of the cell apoptosis by transmission electron microscopy and fluorescence microscopy. Results The cells in groups B and C exhibited reduced viability and signs of apoptosis, and necrosis was observed in group D. Transmission electron microscopy and fluorescence microscopy identified early apoptotic changes and the presence of apoptotic body in the macrophages. The apoptotic rates of groups B and C were much higher than that in group A found by flow cytometry. Conclusion FK506 can promote the apoptosis of macrophage activated by allogenic nerve homogenate and reduce macrophage-mediated immunological rejection of peripheral nerve allograft.

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