乳杆菌改善幽门螺杆菌感染性胃炎的实验研究  

The experimental study of lactobacillus improving gastritis induced by Helicobacter pylori

作　　者：蒋杰[1] 黄煌[1] 梅璐[1] 米阳 刘思濛 孙向东 郑鹏远[1] Jiang Jie;Huang Huang;Mei Lu;Mi Yang;Liu Simeng;Sun Xiangdong;Zheng Pengyuan(The Fifth Affiliated Hospital of Zhengzhou University,Zhengzhou 450052,China)

机构地区：[1]郑州大学第五附属医院,河南郑州450052

出　　处：《河南医学研究》2018年第16期2887-2892,共6页Henan Medical Research

基　　金：国家自然科学基金资助项目(81370494);国家科技支撑计划基金资助项目(2012BAI35B03);郑州市创新型科技人才队伍建设工程基金资助项目(131PLJRC656)

摘　　要：目的探讨乳杆菌在幽门螺杆菌(Hp)相关性胃炎中的作用机制。方法将40只Balb/c小鼠随机分为空白组和实验组(Hp模型组、鼠李糖乳杆菌DM9054+Hp组、植物乳杆菌86066+Hp组)。采用Hp菌株灌胃2周建立小鼠Hp感染模型,造模结束后,实验组中的鼠李糖乳杆菌DM9054+Hp组、植物乳杆菌86066+Hp组分别给予鼠李糖乳杆菌DM9054及植物乳杆菌86066 (1×109CFU/ml,0. 8 ml),且连续灌胃10 d,而空白组及Hp模型组给予等量PBS连续灌胃10 d。干预结束后,使用快速尿素酶法和胃黏膜组织培养来确定各组小鼠胃内有无Hp定植并检测相应细菌数量;采用胃黏膜组织HE染色了解炎症浸润情况。利用Hp和鼠李糖乳杆菌DM9054或植物乳杆菌86066共感染人胃腺癌细胞系(AGS),采用ELISA方法测定细胞培养上清中白细胞介素-8(IL-8)表达水平;采用Real-time PCR法测定IL-8mRNA表达水平;采用Western blot检测MAPK,JAK/STAT,NF-κB炎症通路中的相关蛋白的表达水平。结果在动物实验中,实验组小鼠胃中均有Hp的定植,并且引起了胃黏膜组织炎症。在细胞实验中,Hp感染导致AGS细胞上清中IL-8蛋白上调与细胞中IL-8 mRNA水平升高。鼠李糖乳杆菌DM9054或植物乳杆菌86066的干预抑制了细胞上清液中IL-8蛋白和IL-8 mRNA的表达水平,并下调了磷酸化的ERK1/2、JNK1/2、P38及磷酸化的JAK1、JAK2和STAT3和NF-κB p65蛋白的表达水平(P <0. 05)。结论鼠李糖乳杆菌DM9054和植物乳杆菌86066可能通过下调MAPK、JAK/STAT、NF-κB途径中磷酸化蛋白的表达水平,抑制炎症因子IL-8的释放,从而起到抑制Hp感染引起胃炎的作用。Objective To explore the specific mechanisms of lactobacillus in the treatment of Helicobacter pylori （Hp） in- duced gastric ilfflammation. Methods Forty Blab/c mice were randomly divided into control group and test groups （ Hp group, L. rhamnous DM9054 ＋ Hp group, L. plantarum 86066 ＋ Hp group）. Balb/c mice of Hp ilffection was modeled by intragastric administration for two weeks. L. rhamnous DM9054 ＋ Hp group and L. plantarum 86066 ＋ Hp group were administered with （ 1 x 10^9 CFU/ml, 0. 8 ml）probiotics for 10 days. Control group and Hp group were given the same volume of PBS for 10 days. The results of Hp coloning in Balb/c mice stomach were evaluated by rapid urease tests （RUT） and the number of Hp was calculated by colony forming units per gram of stomach. The gastric ilfflammation was detected by hematoxylin and eosin staining （ HE staining）. In vitro, the AGS cells were treated with Hp and L. rhamnous or L. plantarum. The amount of IL -8 in cell culture medium was measured by ELISA, and interleukin - 8 （ IL - 8 ） mRNA expression was analyzed by real - time PCR. Total and phosphorylation forms of MAPK, JAK/STAT, and NF - KB pathway were measured by Western blot. Results Hp coloned in the stomach of mice in test groups and caused gastric ilfflammation. Both the levels of IL - 8 in the cell supernatant and RNA levels of IL - 8 in AGS cells increased after Hp infection, and L. rhamnous DM9054 and L. plantarum 86066 suppressed levels of IL - 8 in proteins and mRNA and attenuated the expressions of phosphorylation forms of ERK1/2,JNK1/2, P38 , JAK1 ,JAK2, STAT3 and NF - KB subunit p65 （ P 〈 0. 05 ）. Conclusion L. rhamnous or L. plantarum may inhibit Hp - induced gastritis through the inactivation of DhosDhorvlation forms of the MAPK. Jak/Stat. and NF - KB pathways.

关 键 词：胃炎 幽门螺杆菌 益生菌 白细胞介素-8 

分 类 号：R573.3[医药卫生—消化系统]