Please wait a minute...
浙江大学学报（医学版）
浙江大学学报（医学版）  2013, Vol. 42 Issue (5): 543-549    DOI: 10.3785/j.issn.1008-9292.2013.05.011
论著     
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
张军1，张国新2，陈菲菲1，何邦顺1，叶峰2，潘晓林2
1.南京医科大学附属南京医院消化科，南京市第一医院消化科，江苏 南京 210006；
2.南京医科大学第一附属医院消化科，江苏 南京 210029；
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；
 全文: PDF(1348 KB)   HTML ( ) 
摘要：

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    
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 words: Helicobacter pylori    Lipopolysaccharides    Signal transduction    Epithelial cells    Gastric mucosa    Cells, Cultured
收稿日期: 2013-03-20 出版日期: 2013-09-25
CLC:  R 742.1  
基金资助:

南京医科大学科技发展基金（2011NJMU027）.
通讯作者: 张国新（1964-），男，博士，博士生导师，教授，主任医师，主要从事幽门螺杆菌的基础与临床研究、消化道肿瘤的防治研究；E-mail：guoxinz@njmu.edu.cn   
作者简介: 张军（1971-），男，副主任医师，主要从事消化道肿瘤及幽门螺杆菌的相关基础与临床研究；E-mail:zhangjun711028@126.com
服务　　
把本文推荐给朋友
加入引用管理器
E-mail Alert
RSS
作者相关文章　　
张军
张国新
陈菲菲
何邦顺
叶峰
潘晓林

引用本文:

张军, 张国新, 陈菲菲, 何邦顺, 叶峰, 潘晓林. 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[J]. 浙江大学学报（医学版）, 2013, 42(5): 543-549.

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.

链接本文:

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

[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] 魏振龙,石文贵,陈克明,周建,王鸣刚. 淫羊藿素通过CXCR4/SDF-1信号通路促进小鼠成骨细胞成熟和矿化[J]. 浙江大学学报（医学版）, 2017, 46(6): 571-577.
[2] 张斌斌 等. 抑制哺乳动物雷帕霉素靶蛋白信号通路对慢性脑缺血小鼠认知功能的改善和机制[J]. 浙江大学学报（医学版）, 2017, 46(4): 405-412.
[3] 侯甲福,武蕾蕾,才玉婷,高长久,安锦丹,于忠诚. 锦花清热胶囊抗内毒素作用实验研究[J]. 浙江大学学报（医学版）, 2017, 46(1): 74-79.
[4] 马婷婷,王毅,陈晓倩,赵筱萍. 液相色谱-质谱联用导向的黄葵胶囊肾保护活性物质研究[J]. 浙江大学学报（医学版）, 2017, 46(1): 66-73.
[5] 封盛 等. 糖皮质激素受体信号通路在膀胱癌治疗中的作用研究进展[J]. 浙江大学学报（医学版）, 2016, 45(6): 655-660.
[6] 竺天虹 等. 上皮间充质转化介导子宫内膜异位症发生发展的研究进展[J]. 浙江大学学报（医学版）, 2016, 45(4): 439-445.
[7] 陈子博等. 舒尼替尼通过调控转化生长因子β介导的上皮-间质转化抑制卵巢癌细胞转移[J]. 浙江大学学报（医学版）, 2015, 44(5): 479-485.
[8] 辛廖冰, 江秀秀, 叶小磊, 吴瑞瑾, 徐开红, 马俊彦, 林俊. 稳定沉默水通道蛋白5基因对异位子宫内膜腺上皮细胞增殖及迁移的影响[J]. 浙江大学学报（医学版）, 2015, 44(3): 285-292.
[9] 刘志娴, 魏尔清, 卢韵碧. 上皮—间质转化在肿瘤发生发展中的作用研究进展[J]. 浙江大学学报（医学版）, 2015, 44(2): 211-216.
[10] 张春阳, 祝艳, 冯华松, 陈旭昕. 放射线照射的肺成纤维细胞对人脐带间充质干细胞中经典Wnt/β-catenin通路的影响[J]. 浙江大学学报（医学版）, 2015, 44(2): 162-166.
[11] 韩艳霞, 尤良顺, 刘辉, 毛莉萍, 叶琇锦, 钱文斌. 细胞周期蛋白依赖激酶抑制剂诱导HL-60细胞凋亡及分子机制研究[J]. 浙江大学学报（医学版）, 2015, 44(2): 174-178.
[12] 郑永霞，张成文，惠斌，等. 磷脂酰肌醇3激酶调节亚基基因对HepG2细胞增殖的影响[J]. 浙江大学学报（医学版）, 2014, 43(5): 559-.
[13] 赵亮，等. 结直肠癌转移机制研究进展[J]. 浙江大学学报（医学版）, 2014, 43(4): 486-493.
[14] 韩晓瑜，章璐，刘志娴，等. 白三烯D4在体外对A549肺泡上皮细胞增殖及迁移的影响[J]. 浙江大学学报（医学版）, 2014, 43(3): 287-292.
[15] 石文贵，马小妮，陈克明. 初级纤毛在细胞信号转导中的作用与机制[J]. 浙江大学学报（医学版）, 2014, 43(3): 359-365.