Protective effect of syndecan-1 medicated
Bifidobacterium on necrotizing enterocolitis in rats- Journal of Third Military Medical University Vol. 39, Issue 15, Pages: 1549-1555(2017)
- 作者机构：
400014 重庆，重庆医科大学附属儿童医院新生儿科，儿童发育疾病研究省部共建教育部重点实验室，儿科学重庆市重点实验室，重庆市儿童发育重大疾病诊治与预防国际科技合作基地
- 作者简介：
- 基金信息：the National Natural Science Foundation of China(30200300);the National Clinical Key-discipline Construction Program of China(卫办医政(2011)873)
DOI：10.16016/j.1000-5404.201701137
CLC： R-332;R378.994;R574.5Received：24 January 2017，
Revised：2017-04-07，
Published：15 August 2017
- 稿件说明：
移动端阅览
Yifeng LIU, Ling XIANG, Weixia DU, et al. Protective effect of syndecan-1 medicated
Bifidobacterium on necrotizing enterocolitis in rats[J]. Journal of Third Military Medical University, 2017, 39(15): 1549-1555.Yifeng LIU, Ling XIANG, Weixia DU, et al. Protective effect of syndecan-1 medicated
Bifidobacterium on necrotizing enterocolitis in rats[J]. Journal of Third Military Medical University, 2017, 39(15): 1549-1555. DOI： 10.16016/j.1000-5404.201701137.
摘要
目的
2
探索Syndecan-1蛋白在新生大鼠坏死性小肠结肠炎(necrotizing enterocolitis，NEC)模型肠组织中的表达情况，明确双歧杆菌对Syndecan-1蛋白的修复作用。
方法
2
建立新生大鼠NEC模型。按随机数字表法将新生大鼠分为3组(
n
=10)：健康对照组、NEC组和双歧杆菌组。HE染色观察回盲部肠组织病理学变化。qRT-PCR技术检测各组TNF-α、IL-1β、MMP-7和Syndecan-1 mRNA的表达水平。Western blot检测分析各组Syndecan-1蛋白的表达水平。免疫组化分析各组Syndecan-1蛋白的定位表达。
结果
2
NEC组TNF-α、IL-1β和MMP-7 mRNA表达水平高于健康对照组，而双歧杆菌可以抑制TNF-α、IL-1β和MMP-7的表达。NEC组Syndecan-1 mRNA的表达水平略高于健康对照组，双歧杆菌组略低于NEC组，但差异均无统计学意义。Western blot结果显示，健康对照组Syndecan-1蛋白呈高水平表达，NEC组几乎未检测到Syndecan-1蛋白的表达，而双歧杆菌组可见Syndecan-1蛋白的表达。免疫组化结果显示，对照组绒毛上皮细胞膜呈深棕色，而NEC组绒毛上皮细胞膜棕色染色明显减弱，双歧杆菌组上皮细胞膜着浅棕色。
结论
2
NEC炎症时，Syndecan-1蛋白被切割，双歧杆菌可保护Syndecan-1蛋白的表达。双歧杆菌对肠黏膜的保护作用可能是通过抑制MMP-7等因子的表达，从而保护了Syndecan-1蛋白。
Abstract
Objectives
2
To explore the expression of syndecan-1 in the rat model of necrotizing enterocolitis (NEC)
and confirm the recovery effect of
Bifidobacterium
on syndecan-1.
Methods
2
Rat model of NEC was established.A total of 30 Sprague-Dawley rats were randomly divided into 3 groups at the first day after birth
health control group
NEC group and
Bifidobacterium
intervention group
with 10 rats in each group. HE staining was carried out to observe the histological changes in the intestinal tissue of ileocecal segment. The mRNA expression levels of TNF-α
IL-1β
MMP-7 and syndecan-1 were measured by qRT-PCR. The protein expression level of syndecan-1 was detected by Western blotting
and its cellular localization on the intestine was observed through immunohistochemical staining.
Results
2
The mRNA levels of TNF-α
IL-1β and MMP-7 were significantly up-regulated in the NEC group than the normal control.
Bifidobacterium
intervention reduced the mRNA levels of above molecules. The mRNA level of syndecan-1 was mildly higher in the NEC group than the healthy control group
but lower than in the
Bifidobacterium
group
though no obvious differences. Syndecan-1 protein was highly expressed in the healthy rats
not expressed in the NEC rats
and expressed in the intervention rats. The results of immunohistochemical staining showed that syndecan-1 protein were mainly located on the membrane of intestinal epithelial cells in the healthy control group
but mildly in the NEC group and the intervention group
which consistent with the results of Western blotting.
Conclusions
2
Shedding of syndecan-1 happens in the process of NEC inflammation
and
Bifidobacterium
could protect its protein expression. The protective effect of
Bifidobacterium
on intestinal mucosal may be through inhibiting MMP-
7 to repair the syndecan-1 protein.
关键词
Keywords
references
MAHESHWARI A, LACSON A, LU W, et al . Interleukin-8/CXCL8 forms an autocrine loop in fetal intestinal m ucosa[J ] . Pediatr Res, 2004, 56(2): 240-249. DOI:10.1203/01.PDR.0000133196.25949.98.
DAIG R, ROGLER G, ASCHENBRENNER E, et al . Human intestinal epithelial cells secrete interleukin-1 receptor antagonist and interleukin-8 but not interleukin-1 or interleukin-6[J ] . Gut, 2000, 46(3): 350-358. DOI:10.1136/gut.46.3.350.
LI Q, PARK P W, WILSON C L, et al . Matrilysin shedding of syndecan-1 regulates chemokine mobilization and transepithelial efflux of neutrophils in acute lung injury[J ] . Cell, 2002, 111(5): 635-646. DOI:10.1016/s0092-8674(02)01079-6.
GUAN P P, YU X, GUO J J, et al . By activating matrix metalloproteinase-7, shear stress promotes chondrosarcoma cell motility, invasion and lung colonization[J ] . Oncotarget, 2015, 6(11): 9140-9159. DOI:10.18632/oncotarget.3274.
BIN-NUN A, BROMIKER R, WILSCHANSKI M, et al . Oral probiotics prevent necrotizing enterocolitis in very low birth weight neonates[J ] . J Pediatr, 2005, 147(2): 192-196. DOI:10.1016/j.jpeds.2005.03.054.
向玲, 杜维霞, 刘依枫, 等.双歧杆菌通过zonulin影响坏死性小肠结肠炎新生大鼠模型肠道通透性[J].第三军医大学学报, 2016, 38(5): 469-475. DOI:10.16016/j.1000-5404.201507127.
XIANG L, DU W X, LIU Y F, et al . Effect of bifidobacterium on intestinal permeability and expression of zonulin in neonatal rats with necrotizing enterocolitis[J ] . J Third Mil Med Univ, 2016, 38(05):469-475. DOI:10.16016/j.1000-5404.201507127.
李金纯, 韦红, 贾盛华, 等.新生儿坏死性小肠结肠炎动物模型建立方法改进与比较[J].重庆医科大学学报, 2009, 34(3): 313-317.
LI J C, WEI H, JIA S H, et al . Improvement and comparison of animal models of neonatal necrotizing enterocolitis[J ] . J Chongqing Med Univ, 2009, 34(3): 313-317.
魏小娣, 韦红, 贾盛华, 等.姜黄素对新生大鼠坏死性小肠结肠炎模型NF-κB, TNF-α及IL-6表达的影响[J].第四军医大学学报, 2009(22):2537-2540.
WEI X D, WEI H, JIA S H, et al . Effect of curcumin on expression of nuclear factor-κB, tumor necrosis facror-α and interleukin-6 in necrotizing enterocolitis rat model[J ] . J Fourth Mil Med Univ. 2009, 30(22):2537-2540.
AUESTAD N, KORSAK R A, BERGSTROM J D, et al . Milk-substitutes comparable to rat's milk; their preparation, composition and impact on development and metabolism in the artificially reared rat[J ] . Br J Nutr, 1989, 61(3):495-518.
MATSUNO K, ADACHI Y, YAMAMOTO H, et al . The expression of matrix metalloproteinase matrilysin indicates the degree of inflammation in ulcerative colitis[J ] . J Gastroenterol, 2003, 38(4): 348-354. DOI:10.1007/s005350 300062.
FITZGERALD M L, WANG Z, PARK P W, et al . Shedding of syndecan-1 and -4 ectodomains is regulated by multiple signaling pathways and mediated by a TIMP-3-sensitive metalloproteinase[J ] . J Cell Biol, 2000, 148(4): 811-824. DOI:10.1083/jcb.148.4.811.
HOOPER N M, KARRAN E H, TURNER A J. Membrane protein secretases[J]. Biochem J, 1997, 321 (Pt 2): 265-279. DOI:10.1042/bj3210265.
MARSHALL L J, RAMDIN L S, BROOKS T, et al . Plasminogen activator inhibitor-1 supports IL-8-mediated neutrophil transendothelial migration by inhibition of the constitutive shedding of endothelial IL-8/heparan sulfate/syndecan-1 complexes[J ] . J Immunol, 2003, 171(4): 2057-2065. DOI:10.4049/jimmunol.171.4.2057.
EDELSON M B, BAGWELL C E, ROZYCKI H J. Circulating pro-and counterinflammatory cytokine levels and severity in necrotizing enterocolitis[J]. Pediatrics, 1999, 103(4 Pt 1): 766-771. DOI:10.1542/peds.103.4.766.
BENKOE T, RECK C, GLEISS A, et al . Interleukin 8 correlates with intestinal involvement in surgically treated infants with necrotizing enterocolitis[J ] . J Pediatr Surg, 2012, 47(8): 1548-1554. DOI:10.1016/j.jpedsurg.2011.11.049.
WANG X, ZUO D, CHEN Y, et al . Shed Syndecan-1 is involved in chemotherapy resistance via the EGFR pathway in colorectal cancer[J ] . Br J Cancer, 2014, 111(10): 1965-1976. DOI:10.1038/bjc.2014.493.
RAVANTI L, KÄHÄRI V M. Matrix metalloproteinases in wound repair (review)[J]. Int J Mol Med, 2000, 6(4): 391-407.
BAKER E A, LEAPER D J. Profiles of matrix metalloproteinases and their tissue inhibitors in intraperitoneal drainage fluid: relationship to wound healing[J]. Wound Repair Regen, 2003, 11(4): 268-274. DOI:10.1046/j.1524-475x.2003.11406.x.
SCHUPPAN D, FREITAG T. Fistulising Crohn's disease: MMPs gone awry[J]. Gut, 2004, 53(5): 622-624. DOI:10.1136/gut.2003.034207.
VISSE R, NAGASE H. Matrix metalloproteinases and tissue inhibitors of metalloproteinases: structure, function, and biochemistry[J]. Circ Res, 2003, 92(8): 827-839. DOI:10.1161/01.RES.0000070112.80711.3D.
BISTER V, SALMELA M T, HEIKKILÄ P, et al . Matrilysins-1 and -2 (MMP-7 and -26) and metalloelastase (MMP-12), unlike MMP-19, are up-regulated in necrotizing enterocolitis[J ] . J Pediatr Gastroenterol Nutr, 2005, 40(1): 60-66. DOI:10.1097/00005176-200501000-00011.
YU W H, WOESSNER J F. Heparan sulfate proteoglycans as extracellular docking molecules for matrilysin (matrix metalloproteinase 7)[J]. J Biol Chem, 2000, 275(6): 4183-4191. DOI:10.1074/jbc.275.6.4183.
CAPLAN M S, SUN X M, HSEUH W, et al . Role of platelet activating factor and tumor necrosis factor-alpha in neonatal necrotizing enterocolitis[J ] . J Pediatr, 1990, 116(6): 960-964. DOI:10.1016/s0022-3476(05)80661-4.
KLEIN R D, BORCHERS A H, SUNDARESHAN P, et al . Interleukin-1beta secreted from monocytic cells induces the expression of matrilysin in the prostatic cell line LNCaP[J ] . J Biol Chem, 1997, 272(22): 14188-14192. DOI:10.1074/jbc.272.22.14188.
MAUVIEL A. Cytokine regulation of metalloproteinase gene expression[J]. J Cell Biochem, 1993, 53(4): 288-295. DOI:10.1002/jcb.240530404.
BODE L, SALVESTRINI C, PARK P W, et al . Heparan sulfate and syndecan-1 are essential in maintaining murine and human intestinal epithelial barrier function[J ] . J Clin Invest, 2008, 118(1): 229-238. DOI:10.1172/JCI32335.
DAY R M, MITCHELL T J, KNIGHT S C, et al . Regulation of epithelial syndecan-1 expression by inflammatory cytokines[J ] . Cytokine, 2003, 21(5): 224-233. DOI:10.1016/s1043-4666(03)00091-7.
0
Views
0
下载量
0
CSCD
- Meta-XMLDownload
- BibTeXDownload
- EndnoteDownload
- RefMan Download
- NoteExpressDownload
- NoteFirstDownload
Publicity Resources
Related Articles
Related Author
Related Institution
- Technical support is provided by Beijing Founder electronics co., LTD 京ICP备09064830号-19
京公网安备11010802024621 - It is recommended to read the content of this site in Chrome&IE9+. Please switch to extreme mode in browser 360.
- Cookies We use cookies to help provide and enhance our service and tailor content. By continuing, you agree to the use of cookies.
