[1]钟河江,周建云,蒋建新,等.X盒结合蛋白1在低浓度皮质酮对巨噬细胞免疫功能调控中的作用[J].第三军医大学学报,2009,31(20):1964-1968.
 ZHONG He-jiang,ZHOU Jian-yun,JIANG Jian-xin,et al.Role of XBP1 in immune function regulation of macrophage by low concentration of corticosterone[J].J Third Mil Med Univ,2009,31(20):1964-1968.
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X盒结合蛋白1在低浓度皮质酮对巨噬细胞免疫功能调控中的作用(/HTML )
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《第三军医大学学报》[ISSN:1000-5404/CN:51-1095/R]

卷:
31卷
期数:
2009年第20期
页码:
1964-1968
栏目:
论著
出版日期:
2009-10-30

文章信息/Info

Title:
Role of XBP1 in immune function regulation of macrophage by low concentration of corticosterone
作者:
钟河江周建云蒋建新王海燕杨策严军刘庆程英
第三军医大学大坪医院野战外科研究所第四研究室,全军交通医学研究所,全军交通伤重点实验室,创伤、烧伤与复合伤国家重点实验室
Author(s):
ZHONG He-jiang ZHOU Jian-yun JIANG Jian-xin WANG Hai-yan YANG Ce YAN Jun LIU Qing CHENG Ying
State Key Laboratory of Trauma, Burns and Combined Injury, Department 4, Institute of Surgery Research, Daping Hospital, Third Milita-ry Medical University, Chongqing 400042, China
关键词:
糖皮质激素巨噬细胞RNA干扰XBP1免疫功能
Keywords:
glucocorticoids macrophage RNA interference X-box binding protein 1 immune function
分类号:
R341;R392.12;R394.3
文献标志码:
A
摘要:
目的   探讨X盒结合蛋白1 (X-box binding protein 1, XBP1)在低浓度皮质酮对小鼠腹腔巨噬细胞免疫功能调控中的作用。   方法   分离培养成年健康雄性C57BL/6小鼠腹腔巨噬细胞,应用XBP1-RNAi慢病毒或阴性对照慢病毒感染小鼠腹腔巨噬细胞,采用荧光显微镜观察感染效率,感染3、5 d后收集巨噬细胞提取总RNA,应用RT-PCR检测小鼠腹腔巨噬细胞XBP1 mRNA表达水平和干扰效率。XBP1-RNAi慢病毒或阴性对照慢病毒感染离体培养的小鼠腹腔巨噬细胞3 d后,再应用低浓度皮质酮(50 ng/ml)处理小鼠腹腔巨噬细胞1 h,分别应用ELISA法及激光共聚焦显微镜检测小鼠腹腔巨噬细胞培养上清中TNF-α生成及其吞噬功能变化。   结果   当复感染指数(multiplicity of infection, MOI)值≥6时,XBP1-RNAi慢病毒能有效地感染小鼠腹腔巨噬细胞,感染效率>75%。应用RT-PCR检测发现XBP1-RNAi能有效地抑制小鼠腹腔巨噬细胞XBP1基因表达。通过选择性地沉默XBP1基因表达,低浓度皮质酮(50 ng/ml)对巨噬细胞吞噬功能增强的作用受到抑制,巨噬细胞的吞噬率和吞噬指数均显著下降(P<0.01),而且对TNF-α分泌增强的作用也受到抑制,显著低于阴性对照慢病毒组(P<0.01)。   结论   采用RNA干扰选择性地沉默XBP1,能有效地抑制低浓度皮质酮对巨噬细胞吞噬和TNF-α生成增强的作用。
Abstract:
Objective   To explore the role of X-box binding protein 1 (XBP1) in immune function regulation of macrophage by low conceatration of corticosterone.    Methods   Peritoneal macrophages were isolated from adult male C57BL/6 mice, and then infected with XBP1-RNAi lentivirus or negative control lentivirus. The infection efficiency was observed by fluorescence microscopy. Mice peritoneal macrophages were collected in 3 and 5 d after infection and total RNA were extracted. RT-PCR analysis was performed to detect the effect of RNA interference. Furthermore, mice peritoneal macrophages were cultured in vitro and infected with XBP1-RNAi lentivirus or negative control lentivirus for 3 d, and then treated with low concentration of corticosterone (50 ng/ml) for 1 h. TNF-α concentration in the cell culture supernatant and phagocytosis of macrophages were determined by enzyme-linked immunosorbent assay (ELISA) and confocal microscopy, respectively.    Results   When the multiplicity of infection (MOI) was more than 6, the infection efficiency of XBP1-RNAi lentivirus was above 75% after infection 4 d. RT-PCR analysis confirmed that XBP1-RNAi lentivirus effectively inhibited the expression of XBP1 mRNA in mice peritoneal macrophages. Furthermore, after selectively silencing of the XBP1 gene, the enhancement of macrophage phagocytosis by low concentration of corticosterone was efficiently inhibited, phagocytic rate and phagocytic index of mice peritoneal macrophages were decreased markedly compared with that of negative control lentivirus group (P<0.01). Meanwhile, the enhancement of TNF-α production by low concentration of corticosterone was also depressed, the concentration of TNF-α in the cell culture supernatant was significantly declined in XBP-RNAi lentivirus group, compared with the negative control lentivirus group (P<0.01).    Conclusion   Selective silencing of XBP1 by RNA interference may effectively inhibit the enhancement of macrophage phagocytosis and TNF-α production by low concentration of corticosterone.

参考文献/References:

钟河江, 周建云, 蒋建新, 等. X盒结合蛋白1在低浓度皮质酮对巨噬细胞免疫功能调控中的作用[J]. 第三军医大学学报,2009,31(20):1964-1968.

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更新日期/Last Update: 2009-10-21