Please wait a minute...
Journal of ZheJiang University(Medical Science)  2013, Vol. 42 Issue (5): 543-549    DOI: 10.3785/j.issn.1008-9292.2013.05.011
    
Effect of Helicobacter Pylori lipopolysaccharide on expression of Gli and Ptch-1 proteins in sonic hedgehog signaling pathway of gastric mucosa GES-1 cells
ZHANG Jun1, ZHANG Guo-Xin2, CHEN Fei-Fei1, HE Bang-Shun1, YE Feng2, PAN Xiao-Lin2
1.Department of Gastroenterology, Nanjing First Hospital,Nanjing Medical University,Nanjing 210006,China; 2.Department of Gastroenterology, The First Affiliated Hospital of Nanjing Medical University,Nanjing 210029,China;
Download: HTML (   PDF(1348KB)
Export: BibTeX | EndNote (RIS)      

Abstract  
Objective: To investigate the effect of Helicobacter Pylori lipopolysaccharide (Hp-LPS) on expression of Gli and Ptch-1 proteins in sonic hedgehog (Shh) signaling pathway of gastric mucosa GES-1 cells.
Methods: The LPS was extracted from Hp by hot phenol water method, and then the concentration of LPS was detected by the kinetic turbidimetric assay. GES-1 cells were stimulated by different concentrations of Hp-LPS (0, 1, 10, 20, 30 and 40 μg/ml). The inhibition rates of cell growth were measured by MTT assay after treated with Hp-LPS for 24 h. The expression of Gli and Ptch-1 proteins were determined by Western Blot.
Results: MTT assay showed that the inhibition rates of GES-1 cell growth after treatment by different concentrations of Hp-LPS (1, 10, 20, 30 and 40μg/ml) were 25.8%±2.7%, 34.2%±3.1%, 46.3%±3.4%, 60.8%±2.1% and 82.9%±2.8% respectively (r=0.985, P<0.001). Western blot showed that the expressions of Gli and Ptch-1 proteins were decreased after Hp-LPS treatment (0, 1, 10, 20, 30 and 40 μg/ml): the relative expression values of Gli were 1.286±0.180, 0.963±0.067, 0.850±0.085, 0.566±0.058, 0.549±0.056 and 0.377±0.047, respectively (r=-0.945, P<0.001); those of Ptch-1 were 1.688±0.088, 1.466±0.061, 1.170±0.065, 1.042±0.064, 0.648±0.057 and 0.482±0.074, respectively (r=-0.985, P<0.001).
Conclusion: Hp-LPS can decrease the related protein expression of Shh signaling pathway, which indicates that Hp may interfere with the function of Shh signaling pathway in gastric mucosa via the effect of its LPS.


Key wordsHelicobacter pylori      Lipopolysaccharides      Signal transduction      Epithelial cells      Gastric mucosa      Cells, Cultured     
Received: 20 March 2013      Published: 25 September 2013
CLC:  R 742.1  
Cite this article:

ZHANG Jun, ZHANG Guo-Xin, CHEN Fei-Fei, HE Bang-Shun, YE Feng, PAN Xiao-Lin. Effect of Helicobacter Pylori lipopolysaccharide on expression of Gli and Ptch-1 proteins in sonic hedgehog signaling pathway of gastric mucosa GES-1 cells. Journal of ZheJiang University(Medical Science), 2013, 42(5): 543-549.

URL:

http://www.zjujournals.com/med/10.3785/j.issn.1008-9292.2013.05.011     OR     http://www.zjujournals.com/med/Y2013/V42/I5/543


幽门螺杆菌脂多糖对胃粘膜shh信号通路相关蛋白Gli、Ptch-1表达的影响

目的:观察幽门螺杆菌(Helicobacter Pylori,Hp)脂多糖刺激下胃粘膜细胞Shh信号通路发生的变化。
方法:应用热酚水法提取Hp脂多糖,动态浊度法检测脂多糖浓度,将人胃粘膜上皮细胞系GES-1进行细胞培养后分成6组,分别以浓度为0 μg/ml、1 μg/ml、10 μg/ml、20 μg/ml、30 μg/ml、40 μg/ml的Hp脂多糖进行刺激。刺激24 h后,应用MTT法观察各组细胞活性,Western Blot法对各组细胞中Shh信号通路相关蛋白Gli、Ptch-1的表达水平进行检测。
结果:不同浓度Hp脂多糖刺激24h后,MTT法检测结果显示随着Hp脂多糖浓度增加,Hp脂多糖对GES-1细胞增殖抑制越强,分别为25.8%±2.7%,34.2%±3.1%,46.3%±3.4%,60.8%±2.1%,82.9%±2.8%(r=0.985,P<0.001);Western Blot检测结果显示随着Hp脂多糖浓度增加,Gli、Ptch-1表达逐渐下降,Gli相对表达值分别为1.286±0.180、0.963±0.067、0.850±0.085、0.566±0.058、0.549±0.056、0.377±0.047(r=-0.945,P<0.001);Ptch-1相对表达值分别为1.688±0.088、1.466±0.061、1.170±0.065、1.042±0.064、0.648±0.057、0.482±0.074(r=-0.985,P<0.001)。
结论:Hp脂多糖能使Shh信号通路中相关蛋白的表达明显降低,提示Hp对胃粘膜Shh信号通路的影响可能是通过脂多糖实现的。


关键词: 螺杆菌,幽门,  脂多糖类,  信号传导,  上皮细胞,  胃黏膜,  细胞,培养的 
[[1]]   HILDRETH C J,LYNM C,GLASS R M.Helicobacter pylori
[[J]]   JAMA,2008,300(11):1374.
[[2]]   SLOMIANY A.Sucralfate Counteracts the Inhibition of Gasteic Mucosal Ucin Recptor by H.Pylori Lipopolysaccaride
[[J]]   Scand J Fastroenterol,1995,210(30):77-81.
[[3]]   KATOH Y,KATOH M.Hedgehog signaling pathway and gastrointestinal stem cell signaling network
[[J]]   Int J Mol Med,2006,18(6):1019-1023.
[[4]]   NISHIZAWA T,SUZUKI H,NAKAGAWA I,et al.Early Helicobacter pylori eradication restores sonic hedgehog expression in the gastric mucosa of Mongolian gerbils
[[J]]   Digestion,2009,79(2):99-108.
[[5]]   LEE K M,LEE J S,JUNG H S,et al.Late reactivation of sonic hedgehog by Helicobacter pylori results in population of gastric epithelial cells that are resistant to apoptosis:implication for gastric carcinogenesis
[[J]]   Cancer Lett,2010,187(1):44-53.
[[6]]   LUO Yi-hui,YAN Jie,MAO Ya-fei.Helicobacter pylori lipopolysaccharide:Biological activities in vitro and in vivo,pathological correlation to human chronic gastritis and peptic ulcer
[[J]]   World J Gastroenterol,2004,10(14):2055-2059.
[[7]]   DIXON M F.Prospects for intervention in gastric carcinogenesis:reversibility of gastric atrophy and intestinal metaplasia
[[J]]   Gut,2001,49(1):2-4.
[[8]]   MEINING A,MORGNER A,MIEHLKE S,et al.Atrophy-metaplasia-dysplasia-carcinoma sequence in the stomach:a reality or merely an hypothesis?
[[J]]   Best Pract Res Cli Gastroenterol,2001,15(6):983-998.
[[9]]   BHATTACHARYYA A,PATHAK S,DATTA S,et al.Mitogen-activated protein kinases and nuclear factor-kappa B regulate Helicobacter pylori-mediated interleukin-8 release from macrophages
[[J]]   Biochem J,2002,368(Pt 1):121-129.
[[10]]   INNOCENTI M,SVENNERHOLM A M,QUIDING-JRBRINK M.Helicobacter pylori lipopolysaccharides preferentially induce CXC chemokine production in human monocytes
[[J]]   Infect Immun,2001,69(6):3800-3808.
[[11]]   HUANGFU D,ANDERSON KV.Signaling from Smo to Ci/Gli:conser 2 vation and divergence of Hedgehog pathways from Drosophila to vertebrates
[[J]]   Development,2006,133(1):3-14.
[[12]]   MARTIN J,DONNELLY JM,HOUGHTON J,et al.The Role of sonic hedgehog reemergence during gastric cancer
[[J]]   Dig Dis Sci,2010,55(6):1516-1524.
[[13]]   SUZUKI H,MINEGISHI Y,NOMOTO Y,et al.Down-regulation of a morphogen (sonic hedgehog) gradient in the gastric.epithelium of Helicobacter pylori-infected Mongolian gerbils
[[J]]   J Pathol,2005,206 (2):186-197.
[[14]]   MA X L,SUN H J,WANG Y S,et al.Study of Sonic hedgehog signaling pathway related molecules in gastric carcinoma
[[J]]   World J Gastroenterol,2006,12(25):3965-3969.
[[15]]   PIOTROWSKI J,PZOTROWSKI E,SKRODZAK D,et al.Induction of acute gastritis and epithelial apoptosis by Helicobacter pyloric lipopoly saccharide
[[J]]   Scand J Gustroenterol,1997,32:203-211.
[[16]]   SMITH MF J R,MITCHELL A,LI G,et al.Toll-like Receptor (TLR) 2 and TLR5,but Not TLR4,are required for Helicobacter pylori-induced NF-kappaB activation and chemokine expression by epithelial cells
[[J]]   J Biol Chem,2003,278(35):32552-32560.
[[17]]   KIM J H,CHOI Y J,LEE S H,et al.Effect of Helicobacter pylori infection on the sonic hedgehog signaling pathyway in gastric cancer cell
[[J]]   Oncol Rep,2010,23(6):1523-1528.
[[18]]   YANG Y,LI Q,DENG Z,et al.Protection from lipopolysaccharide-induced pulmonary microvas-cular endothelial cell injury by activation of hedgehog signaling pathway
[[J]]   Mol Biol Rep,2011,38(6):3615-3622.
[[19]]   LEE S Y,HAN H S,LEE K Y,et al.Sonic hedgehog expression in gastric cancer and gastric adenoma
[1] WEI Zhenlong,SHI Wengui,CHEN Keming,ZHOU Jian,WANG Minggang. Icaritin promotes maturation and mineralization of mouse osteoblast MC3T3-E1 cells through CXCR4/SDF-1 signal pathway[J]. Journal of ZheJiang University(Medical Science), 2017, 46(6): 571-577.
[2] ZHANG Binbin, WU Meiling, LIU Luna, ZHU Yangbin, KAI Jiejing, ZENG Linghui. Inhibiting mammalian target of rapamycin signaling pathway improves cognitive function in mice with chronic cerebral ischemia[J]. Journal of ZheJiang University(Medical Science), 2017, 46(4): 405-412.
[3] HE Yujie,PAN Jianping. Progress on mechanisms for pathogensto evade NOD-like receptor and Toll-like receptor signaling pathways[J]. Journal of ZheJiang University(Medical Science), 2017, 46(2): 218-224.
[4] HOU Jiafu,WU Leilei,CAI Yuting,GAO Changjiu,AN Jindan,YU Zhongcheng. Antiendotoxin effect of Jinhuaqingre capsules[J]. Journal of ZheJiang University(Medical Science), 2017, 46(1): 74-79.
[5] MA Tingting,WANG Yi,CHEN Xiaoqian,ZHAO Xiaoping. LC/MS guided approach to discovering nephroprotective substances from Huangkui capsule[J]. Journal of ZheJiang University(Medical Science), 2017, 46(1): 66-73.
[6] FENG Sheng, CHEN Jijun, ZHENG Yichun. Research progress on the effect of glucocorticoid receptor signaling pathways in bladder cancer[J]. Journal of ZheJiang University(Medical Science), 2016, 45(6): 655-660.
[7] ZHU Tianhong, ZHANG Xinmei. Research progress on the role of epithelial-mesenchymal transition in pathogenesis of endometriosis[J]. Journal of ZheJiang University(Medical Science), 2016, 45(4): 439-445.
[8] XIE Chuan, LYU Nonghua. Kyoto global consensus report for treatment of Helicobacter pylori and its implications for China[J]. Journal of ZheJiang University(Medical Science), 2016, 45(1): 1-4.
[9] CHEN Zi-bo, CHANG Lin-lin, ZHOU Tian-yi, WANG Dan-dan, CHEN Ying, ZHAO Ping-ge, ZHU Hong. Sunitinib suppresses migration of ovarian cancer cells through negative modulation of TGF-β-mediated epithelial-mesenchymal transition[J]. Journal of ZheJiang University(Medical Science), 2015, 44(5): 479-485.
[10] XIN Liao-bing, JIANG Xiu-xiu, YE Xiao-lei, WU Rui-jin, XU Kai-hong, MA Jun-yan, LIN Jun. AQP5 gene silencing inhibits proliferation and migration of ectopic endometrial glandular epithelial cells in endometriosis[J]. Journal of ZheJiang University(Medical Science), 2015, 44(3): 285-292.
[11] LIU Zhi-xian, WEI Er-qing, LU Yun-bi. Research progress on epithelial-mesenchymal transition in cancer recurrence and metastasis[J]. Journal of ZheJiang University(Medical Science), 2015, 44(2): 211-216.
[12] ZHANG Chun-yang, ZHU Yan, FENG Hua-song, CHEN Xu-xin. Effect of irradiated human lung fibroblasts on activation of canonical Wnt/β-catenin signaling pathway in mesenchymal stem cells[J]. Journal of ZheJiang University(Medical Science), 2015, 44(2): 162-166.
[13] HAN Yan-xia, YOU Liang-shun, LIU Hui, MAO Li-ping, YE Xiu-jin, QIAN Wen-bin. Apoptosis of acute myeloid leukemia HL-60 cells induced by CDK inhibitor SNS-032 and its molecular mechanisms[J]. Journal of ZheJiang University(Medical Science), 2015, 44(2): 174-178.
[14] ZHENG Yong-xia, ZHANG Cheng-wen, HUI Bin, et al. Roles of PIK3R1 gene in development of hepatocellular carcinoma[J]. Journal of ZheJiang University(Medical Science), 2014, 43(5): 559-.
[15] ZHAO Liang,et al. Progression in metastasis of colorectal carcinoma[J]. Journal of ZheJiang University(Medical Science), 2014, 43(4): 486-493.