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生长抑素诱导猕猴外周血中性粒细胞凋亡
http://www.100md.com 2005年3月1日 《世界华人消化杂志》 2005年第5期
     许兰涛, 唐承薇, 四川大学华西医院消化内科 四川省成都市 610041

    许兰涛,男, 1964-07-03生, 副主任医师, 副教授, 博士, 主要从事消化内科的研究.

    国家自然科学基金资助项目, No. 30170875

    项目负责人: 唐承薇, 610041, 四川省成都市, 四川大学华西医院消化内科. sccdlx@sohu.com

    电话: 028-85445341

    收稿日期: 2004-12-22 接受日期: 2005-01-20

    Effect of somstostatin on apoptosis of polymorphonuclear neutrophils of rhesus monkey

    Lan-Tao Xu, Cheng-Wei Tang

    Lan-Tao Xu, Cheng-Wei Tang, Department of Digestology, West China Hospital, Sichuan University, Chengdu 610041, Sichuan Province, China

    Supported by National Science Foundation of China, No.30170875

    Correspondence to: Cheng-Wei Tang, Department of Digestology, West China Hospital, Sichuan University, Chengdu 610041, Sichuan Province, China. sccdlx@sohu.com

    Received: 2004-12-22 Accepted: 2005-01-20

    Abstract

    AIM:
To study the effect of somstostatin on the apoptosis of polymorphonuclear neutrophils (PMNs) in rhesus monkey.

    METHODS: PMNs were isolated from whole blood of normal rhesus monkey using one-step discontinuous Percoll gradient centrifugation technique. PMNs were cultured with 5´10-5mol/L somstostatin. Cell morphology was examined by light and electronic microscopy. Apoptosis was detected by flow cytometry and DNA agarose electrophoresis.

    RESULTS: Somstostatin at 5´10-5mol/L induced significant apoptosis in PMNs. The cells manifested typical apoptotic morphology: cell membrane blebbing, chromatin condensation, nucleic fragmentation and apoptotic body formation. Agarose electrophoresis revealed specific DNA ladder. Flow cytometry analysis showed that the ratio of PMNs undergoing apoptosis was remarkably increased from 22.1% to 50.5% following somstostatin treatment (P<0.01).

    CONCLUSION: Somstostatin induces apoptosis of PMNs in rhesus monkey. It may be a new target for the treatment of multiple organ dysfunction syndrome (MODS).

    Key Words: PMN; Apoptosis; Somatostatin; Rhesus monkey; MODS

    Xu LT, Tang CW. Effect of somstostatin on apoptosis of polymorphonuclear neutrophils of rhesus monkey. Shijie Huaren Xiaohua Zazhi 2005;13(5):617-620

    摘要

    目的:
观察生长抑素对猕猴外周血中性粒细胞的作用.

    方法: 用生长抑素与猕猴外周血中性粒细胞共同孵育一定时间后,采用流式细胞仪、DNA电泳、电镜观察猕猴外周血中性粒细胞的形态变化,细胞DNA含量分布变化.

    结果: 生长抑素能诱导猕猴外周血中性粒细胞凋亡,在形态学上表现为细胞膜起泡、染色质固缩、核碎裂、凋亡小体形成;电泳呈凋亡特征性Ladder带;流式细胞仪分析显示,PMN凋亡比例增高(22.1%vs50.5%. P<0.01).

    结论: 生长抑素能诱导猕猴外周血中性粒细胞凋亡.

    关键词: 猕猴; 生长抑素; 中性粒细胞; 凋亡; 多器官功能衰竭

    许兰涛, 唐承薇. 生长抑素诱导猕猴外周血中性粒细胞凋亡. 世界华人消化杂志 2005;13(5):617-620

    :无凋亡图; B:凋亡.

    1 不同浓度生长抑素对PMN凋亡率的影响


    组别


    凋亡率(%)


    1


    对照组


    22.1±0.28


    2


    ss (5×10-6 mol/L)


    33.6±0.52


    3


    ss (5×10-5 mol/L)


    50.5±0.56


    P<0.01vs对照组.

    

    2.3 DNA电泳结果
将5´108/L PMN细胞与5´10-5 mol/L生长抑素作用30 min,DNA电泳呈明显凋亡带,对照组则无凋亡带(图2).

    2 (PDF) DNA电泳Ladder带.

    2.4 电镜观察 将5´108/L PMN细胞与5´10-5 mol/L生长抑素作用30 min,电镜下可见染色质固缩,聚集于核膜下成块状,胞核固缩,染色质凝集,细胞质浓缩,部分细胞碎裂成片,形成质膜包被的大小不等的凋亡小体(图3).

    图3 中性粒细包超微结构TEM×8000.A: 正常; B:凋亡; C:凋亡小体.

    3 讨论由于分子生物学发展,尤其是抗炎症递质治疗在临床应用中并不理想后,国内外学者转向炎症递质调控凋亡的研究,就凋亡和MODS发生机制作了新的探讨.Haslett[22-26]提出炎症反应的失调与中性粒细胞凋亡比率有关.Keel[27-30]在MODS研究中发现外周血中中性粒细胞(PMN)凋亡延迟,并且血中有抗凋亡因子存在.认为凋亡延迟引起PMN寿命延长可能与MODS发生有关.Goris[31]提出经过特异性细胞因子触发凋亡可以解释进行性过度炎症反应可以发展到MODS的最后假说.

    目前已经明确MODS是过度炎症反应的结果,在创伤后有MODS高风险的患者中,PMN凋亡延迟,血浆中的递质使PMN凋亡延迟,并且PMN的作用引发过度炎症反应及器官损伤,创伤后PMN的凋亡在MODS的发生发展中可能具有重要作用.凋亡区别于坏死的重要之处是凋亡的细胞能被巨噬细胞或周围组织细胞平稳识别、清除而不引发炎症反应,因此有研究认为通过凋亡方式清除炎症部位的PMN是限制组织损伤,促进炎症吸收的重要机制.识别清除凋亡PMN的能力降低,大量凋亡的PMN不能被及时有效地处理而裂解坏死,炎症不易控制,因此这种损伤很容易导致MODS.本实验研究表明:生长抑素能诱导中性粒细胞凋亡,可以减少氧自由基、蛋白酶和炎性递质的释放,有利于巨噬细胞的吞噬,必将为治疗MODS开辟新的领域.

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    编辑 潘伯荣, 百拇医药( 许兰涛,唐承薇)
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