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  

Effect of Helicobacter Pylori lipopolysaccharide on expression of Gli and Ptch-1 proteins in sonic hedgehog signaling pathway of gastric mucosa GES-1 cells

作　　者：张军[1] 张国新[2] 陈菲菲[1] 何邦顺[1] 叶峰[2] 潘晓林[2] 

机构地区：[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

出　　处：《浙江大学学报（医学版）》2013年第5期543-549,共7页Journal of Zhejiang University(Medical Sciences)

基　　金：南京医科大学科技发展基金(2011NJMU027)

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0μg/ml、1μg/ml、10μg/ml、20μg/ml、30μg/ml、40μg/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。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 GES1 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 Ptch1 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-1proteins 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 Pteh-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.

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 

分 类 号：R57[医药卫生—消化系统]