[1]王伍超,陈金萍,张岫竹,等.CRH对腹腔巨噬细胞分泌IL-10的影响及其信号机制[J].陆军军医大学学报(原第三军医大学学报),2009,31(22):2228-2231.
 WANG Wu-chao,CHEN Jin-ping,ZHANG Xiu-zhu,et al.Effects of corticotrophin-releasing hormone on IL-10 secretion from peritoneal macrophages and its signal mechanism[J].J Amry Med Univ (J Third Mil Med Univ),2009,31(22):2228-2231.
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CRH对腹腔巨噬细胞分泌IL-10的影响及其信号机制(/HTML )
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陆军军医大学学报(原第三军医大学学报)[ISSN:1000-5404/CN:51-1095/R]

卷:
31卷
期数:
2009年第22期
页码:
2228-2231
栏目:
论著
出版日期:
2009-11-30

文章信息/Info

Title:
Effects of corticotrophin-releasing hormone on IL-10 secretion from peritoneal macrophages and its signal mechanism
作者:
王伍超陈金萍张岫竹代维刘大维周继红
第三军医大学大坪医院野战外科研究所第四研究室,全军/重庆市交通医学研究所,创伤、烧伤与复合伤国家重点实验室
Author(s):
WANG Wu-chao CHEN Jin-ping ZHANG Xiu-zhu DAI Wei LIU Da-wei ZHOU Ji-hong
State Key Laboratory of Trauma, Burns and Combined Injury, Department 4, Institute of Surgery Research, Daping Hospital, Third Military Medical University,  Chongqing 400042, China
关键词:
促肾上腺皮质激素释放激素巨噬细胞IL-10CRHRcAMP/PKA
Keywords:
corticotrophin-releasing hormone macrophage IL-10 CRHR cAMP/PKA
分类号:
R392-33;R392.12;R977.11
文献标志码:
A
摘要:
目的  研究促肾上腺皮质激素释放激素(corticotrophin-releasing hormone,CRH)对大鼠腹腔巨噬细胞分泌白介素-10(interleukin-10,IL-10)的影响及其信号转导机制。  方法  将原代培养的大鼠腹腔巨噬细胞分为正常对照组、CRH组、CP+CRH组、HL+CRH组、H89+CRH组以及PKAi+CRH组。其中正常对照组仅加入新鲜RPMIl640培养液2 ml;CRH处理组,加入含CRH终浓度为10-9 mol/L的培养液2 ml;后4组在分别用CRHR1选择性阻断剂CP-154526(5×10-4 mol/L)、CRHR1/2非选择性阻断剂α-helical CRH9-41 (10-6 mol/L)、AC抑制剂H89(10-6mol/L)和PKA抑制剂PKAi (6-22) amide(10-4 mol/L)预孵育2 h后,加入含终浓度为10-9 mol/L的CRH培养液,再分别加入CRH(终浓度为109 mol/L)刺激。各组分别于CRH刺激后0.5、1、2、4、8 h收集培养皿中的上清液,应用ELISA方法检测IL-10的含量。  结果  正常对照组巨噬细胞分泌IL-10的含量为(29.41±3.10) μg/g。CRH组IL-10含量呈现双峰,在1、4 h达到高峰,分别为( 53.15±7.98)、(46.01±1.70)μg/g,与CRH组0.5 h相比差异均非常显著(P<0.01),接近正常对照组的2倍(P<0.01)。   CP+CRH组与HL+CRH组IL-10含量在1、4 h均明显低于CRH组(P<0.01),并接近正常对照组水平。但在8 h时明显增加,显著高于正常对照组(P<0.05)和CRH组(P<0.05)。且两组之间IL-10含量在0.5~4 h差异不显著,但在8 h时HL+CRH组显著低于CP+CRH组。   H89预孵育后细胞IL-10含量在1、4、8 h显著下降,与CRH组比较差异显著(P<0.01);在4、8 h低于正常对照组水平(P<0.05)。PKAi (6-22) amide预孵育细胞后,IL-10含量在0.5~8 h下降,与正常对照组以及CRH组比较差异非常显著(P<0.01)。  结论  CRH可通过CRHR1促进巨噬细胞分泌产生抗炎因子IL-10,其与cAMP/PKA信号通路密切相关。
Abstract:
Objective   To investigate the effects of corticotrophin-releasing hormone (CRH) on production of interleukin-10 (IL-10) from macrophages and to further elucidate its signal transduction mechanism.    Methods   Rat peritoneal macrophages cultured were divided into 6 groups: control group, CRH group, CP+CRH group, HL+CRH group, H89+CRH group and PKAi+CRH group. The control group was cultured with 2 ml RPMI1640 medium. The CHR group was stimulated with 2 ml CRH at a final concentration of 10 9 mol/L.The other four groups were respectively pretreated with CRHR1 antagonist CP154526 at 5×10-4 mol/L, CRHR1/2 nonselective antagonist α-helical CRH9-41 at 10-6 mol/L, AC inhibitor H89 compound at 10-6 mol/L or PKA inhibitor PKAi (6-22) amide (10-4 mol/L) for 2 h before stimulation with CRH. The concentration of IL-10 in the supernatant after stimulation for 0.5, 1, 2, 4 and 8 h were measured by ELISA method.    Results   In the normal control group, the average concentration of IL-10 kept at  (29.41±3.10)μg/g. As macrophages exposed to CRH, the content of IL-10 increased quickly and reached its first peak at 1 h compared with that in CRH group at 0.5 h (P<0.01), nearly two folds of that in normal control group (P<0.01), then its level decreased by degrees. There existed a second peak of IL-10 expression at 4 h, which was significantly higher than that in CRH group at 0.5 h (P<0.01) and that in control group (P<0.01). The concentration of IL-10 in CP+CRH group and HL+CRH group at 1 and 4 h were significantly lower than that in CRH group (P<0.01), and were similar to that in control group (P>0.05). At 8 h, the concentration of IL-10 in the 2 groups were significantly higher than that in normal control group (P<0.01) or CRH group (P<0.01). There was no statistical difference of IL-10 concentration between CP+CRH group and HL+CRH group (P>0.05) at 0.5 to 4 h, however  that in HL+CRH group was significantly lower than that in CP+CRH group at 8 h (P<0.01). Furthermore, pretreatment of macrophages with AC inhibitors H89 in H89+CRH group induced a significant inhibition of the content of IL-10 cytokines at 1, 4 and 8 h, while the content of IL-10 cytokines at 4 and 8 h was lower than that in normal control group (P<0.05). After pretreated with PKAi (6-22) amide, the content of IL-10 was significantly inhibited during 0.5 to 8 h compared with that in CRH group or control group(P<0.01).    Conclusion   CRH increases IL-10 production by rat peritoneal macrophages through activating CRHR1, which involves in the intracellular cAMP/PKA signal pathway.

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更新日期/Last Update: 2009-11-23