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| 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； |
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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.
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Received: 20 March 2013
Published: 25 September 2013
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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
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| [[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-JRBRINK 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
|
|
|
|
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