[1]曹乐,王成,申明强,等.电离辐射对小鼠肠道隐窝素4表达的影响及意义[J].第三军医大学学报,2015,37(04):330-334.
 Cao Le,Wang Cheng,Shen Mingqiang,et al.Alteration and significance of small intestinal cryptdin 4 in ionizing-radiated mice[J].J Third Mil Med Univ,2015,37(04):330-334.
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电离辐射对小鼠肠道隐窝素4表达的影响及意义(/HTML )
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《第三军医大学学报》[ISSN:1000-5404/CN:51-1095/R]

卷:
37卷
期数:
2015年第04期
页码:
330-334
栏目:
论著
出版日期:
2015-02-28

文章信息/Info

Title:
Alteration and significance of small intestinal cryptdin 4 in ionizing-radiated mice
作者:
曹乐王成申明强王艾平陈芳耿放陈默张乃心粟永萍程天民王军平
第三军医大学军事预防医学院全军复合伤研究所,防原医学教研室,创伤、烧伤与复合伤国家重点实验室,重庆市纳米医药工程技术研究中心
Author(s):
Cao Le Wang Cheng Shen Mingqiang Wang Aiping Chen Fang Geng Fang Chen Mo Zhang Naixin Su Yongping Cheng Tianmin Wang Junping

State Key Laboratory of Trauma, Burns and Combined Injury, Institute of Combined Injury, Chongqing Engineering Research Center for Nanomedicine, College of Military Preventive Medicine, Third Military Medical University, Chongqing, 400038, China

关键词:
电离辐射隐窝素4抗菌LPSIL-1&beta
Keywords:
ionizing radiation cryptdin 4 antibacterial activity LPS IL-1&beta
分类号:
R322.45;R392.3;R818.02
文献标志码:
A
摘要:

目的      检测小鼠受电离辐射后隐窝素4(cryptdin 4, Crp4)的表达变化,探讨其在电离辐射肠损伤中的生物学意义。      方法      8周龄BALB/c小鼠采用随机数字表法分为两组,未辐照组(n=5)和辐照组(n=15),辐照组给予 8 Gy X射线全身一次性照射,分别于照后9、24、72 h取小鼠小肠组织和眼球血。免疫组织化学染色和Western blot分析小鼠小肠Paneth细胞隐窝素Crp4的表达变化,鲎试剂法检测小鼠血清LPS水平。化学合成Crp4多肽,采用涂布克隆计数法分析Crp4对肠道条件致病菌(大肠杆菌、粪肠球菌)和益生菌(乳酸杆菌、双歧杆菌)的抗菌活性;同时,观察Crp4对LPS诱导促炎因子IL-1β 生成的影响。      结果      与对照组相比,小鼠电离辐射后小肠Crp4表达水平升高,72 h升高尤为显著(P<0.01);辐照小鼠血清LPS含量在9 h后显著增加(P<0.05),72 h后升高最为明显(P<0.01);Crp4对条件致病菌(大肠杆菌、粪肠球菌)的抗菌活性显著强于益生菌(乳酸杆菌、双歧杆菌),并能抑制LPS诱导单核巨噬细胞IL-1β的生成(P<0.01)。      结论      电离辐射可以诱导小鼠肠道隐窝素Crp4的表达上调,Crp4表达上调可发挥调节肠道菌群、减轻机体炎症反应的作用。

Abstract:

Objective      To observe the effect of ionizing radiation on the expression of mouse cryptdin 4 (Crp4) and explore the biological significance.        Methods      BALB/c mice (8 weeks old) were randomly divided into a control group (n=5) and a radiated group (n=15), and the radiated group was treated by total-body ionizing radiation (8 Gy X-ray). The small intestinal tissue and eyeball blood were collected before and after ionizing radiation (at 9, 24, and 72 h respectively). Immunohistochemical assay and Western blot analysis were employed to detect the alteration of Crp4 in mouse small intestine. Quantitative chromogenic tachypleus amebocyte lysate test was  used to analyze the alteration of lipopolysaccharide (LPS) in blood serum. Crp4 peptide was obtained through chemical synthesis. Colony counting was employed to assess the antibacterial activity of Crp4 against Escherichia coli, Enterococcus faecalis, Lactobacillus acidophilus, and Bifidobacterium bifidum. Also, the effect of Crp4 on the release of IL-1β from mouse RAW264.7 cells after LPS simulation was evaluated.        Results      The expression of Crp4 in the mouse small intestine was increased after ionizing radiation, and the alteration was the most remarkable at 72 h (P<0.01). The serum LPS level of the radiated mice was significantly increased after 9 h (P<0.05), and peaked after 72 h (P<0.01). Conditional pathogenic bacteria (Escherichia coli and Enterococcus faecalis) were more sensitive to Crp4 than probiotics (Lactobacillus acidophilus and Bifidobacterium bifidum). Moreover, it was found that Crp4 could decrease the release of IL-1β from RAW264.5 cells stimulated with LPS.       Conclusion      Ionizing radiation can induce the up-regulation of Crp4 expression, which might play an important role in the regulation of the intestinal flora and relieve of the inflammatory responses.

参考文献/References:

[1]Neish A S. Microbes in gastrointestinal health and disease[J]. Gastroenterology, 2009, 136(1): 65-80. 
[2]吴小平, 贺杰. 肠道菌群失调与肠外疾病[J]. 医学新知杂志, 2014, 24(3): 188-189, 194. 
[3]席春晖, 冯志松. 肠道菌群在非酒精性脂肪性肝病发病机制中的研究进展[J]. 医学综述, 2014, 20(11): 1953-1955. 
[4]张英春, 韩雪, 单毓娟, 等. 益生菌抑制致病菌作用的机制研究进展[J]. 微生物学通报, 2012, 39(9): 1306-1313. 
[5]Rosengren K J, Daly N L, Fornander L M, et al. Structural and functional characterization of the conserved salt bridge in mammalian paneth cell-defensins: solution structures of mouse CRYPTDIN-4 and (E15D)-CRYPTDIN-4[J]. J Biol Chem, 2006, 281(38): 28068-28078. 
[6]Ayabe T, Satchell D P, Wilson C L, et al. Secretion of microbicidal alpha-defensins by intestinal Paneth cells in response to bacteria[J]. Nat Immunol, 2000, 1(2): 113-118.
[7]Klotman M E, Chang T L. Defensins in innate antiviral immunity[J]. Nat Rev Immunol, 2006, 6(6): 447-456. 
[8]Crovella S, Antcheva N, Zelezetsky I, et al. Primate beta-defensins--structure, function and evolution[J]. Curr Protein Pept Sci, 2005, 6(1): 7-21. 
[9]Porter E M, Liu L, Oren A, et al. Localization of human intestinal defensin 5 in Paneth cell granules[J]. Infect Immun, 1997, 65(6): 2389-2395.
[10]Satoh Y, Habara Y, Ono K, et al. Carbamylcholine- and catecholamine-induced intracellular calcium dynamics of epithelial cells in mouse ileal crypts[J]. Gastroenterology, 1995, 108(5): 1345-1356. 
[11]Gorbunov N V, Kiang J G. UP-regulation of autophagy in small intestine Paneth cells in response to total-body gamma-irradiation[J]. J Pathol, 2009, 219(2): 242-252. 
[12]刘霞, 梁振明, 杨景云, 等. 60Co辐射后肠道菌群改变的实验研究[J]. 黑龙江医药科学, 2002, 25(3): 10. 
[13]Wilmes M, Cammue B P, Sahl H G, et al. Antibiotic activities of host defense peptides: more to it than lipid bilayer perturbation[J]. Nat Prod Rep, 2011, 28(8): 1350-1358. 
[14]Nuding S, Zabel L T, Enders C, et al. Antibacterial activity of human defensins on anaerobic intestinal bacterial species: a major role of HBD-3[J]. Microbes Infect, 2009, 11(3): 384-393. 
[15]Peschel A. How do bacteria resist human antimicrobial peptides?[J]. Trends Microbiol, 2002, 10(4): 179-186. 
[16]Rumio C, Besusso D, Palazzo M, et al. Degranulation of paneth cells via toll-like receptor 9[J]. Am J Pathol, 2004, 165(2): 373-381. 
[17]Shi J, Aono S, Lu W, et al. A novel role for defensins in intestinal homestasis: regulation of IL-1beta secretion[J]. J Immunol, 2007, 179(2): 1245-1253. 
[18]Ramasundara M, Leach S T, Lemberg D A, et al. Defensins and inflammation: the role of defensins in inflammatory bowel disease[J]. J Gastroenterol Hepatol, 2009, 24(2): 202-208. 
[19]Kelly P, Bajaj-Elliott M, Katubulushi M, et al. Reduced gene expression of intestinal alpha-defensins predicts diarrhea in a cohort of African adults[J]. J Infect Dis, 2006, 193(10): 1464-1470.

更新日期/Last Update: 2015-01-30