In Vitro Investigate The Regulation Of The Lactobacillus From Human On The Helicobacter Pylori-induced Acute Inflammatory Reaction To SGC7901 Cells And The Possible Pathway

Objective:1 To investigate the role of both the lactobacillus from human and ATCC4356on the growth of the Helicobacter pylori Sydney strain 1 (SS1).2 To investigate the adhesive ability to gastric epithelial cell SGC7901 of thelactobacillus and the effects of Helicobacter pylori on their adhesive ability.3 Selection of lactobacillus from human with characteristic of effectiveinhibition ability on Helicobacter pylori.4 To investigate the effect of this lactobacillus from human on the level ofPhosphorylated P38 mitogen-actived protein kinase (P-p38 MAPK) inSGC7901 cells treated by Helicobacter pylori.5 To investigate the expression of the IL-8 induced by Helicobacter pyloriand the effects of this lactobacillus from human on IL-8.Methods:1 SGC7901 cell was cultivated to reach exponential growth phase ,then adjusted concentration (2.5×10~5/ml) to plant six shadow mask and incubationfor 24 hours. Experiment subgroup as follow: normal control, H.pyloristimulate group (HP group),ATCC4356 intervention group(ATeC4356group), lactobacillus from human intervention group(Lac12, Lac15, Lac22,Lac31 group).Until reached exponential growth phase ATCC4356 andlactobacillus from human were cultivated and adjusted to same concentrationto incubation with SGC7901 cell together for one hour, then add H.pylori intothem for 3 hours .Contrast analysis the result.2 Experiment subgroup as follow: normal control, H.pylori stimulategroup(HP group), ATCC4356 intervention group(ATCC4356 group), lactobacillus from human intervention group(Lac 12, Lac 15, Lac22, Lac31 group).3 The well test was used to observe the growth inhibitory of the Helicobacterpylori caused by lactobacillus.4 Gram staining and fluorescent staining were used to detected the adhesiveability to gastric epithelial cell SGC7901 of the lactobacillus and the effectsof Helicobacter pylori on the adhesive ability. The Lac 15 strain were selected,which displaying strong restraining H.pylori growth ability in vitro.5 To investigate the effect of this lactobacillus from human on the level ofPhosphorylated P38 mitogen-actived protein kinase (P-p 38 MAPK) inSGC7901 cells treated by Helicobacter pylori (SS1).6 The release of IL-8 in cells was detected by ELISA.Results:1 The diameter of both lactobacillus from human supematant bacteriostasiscingula was longer than the ATCC4356.2 Both lactobacillus can adhere SGC7901 cell, the adhesion index number of the lactobacillus from human increased compared with ATCC4356 when bothof the lactobacillus concentration was 3×10~9cfu/ml .the adhesion index andcell adhesion rate of Lac22 was the largest. There are significance differencebetween Lac22 and other group. The adhesion index number andlactobacillus adhesion rate of ATCC4356+HP group compared with those ofATCC4356 group, Lac12+HP group compared with those of Lac12group,Lac15+HP group compared with those of Lac15group , Lac22+HP groupcompared with those of Lac22group, Lac31+HP group compared with thoseof Lac31group are significance decreased, fluorescent staining were used todetected the adhesive ability to gastric epithelial cell SGC7901 of thelactobacillus and the effects of Helicobacter pylori on the adhesive ability.The Lac15 strain were selected, which displaying strong restraining H.pylorigrowth ability in vitro.3 The level of phosphorylation of p38MAPK and the level of IL-8 secretoryvolume of SGC7901 cells in HP group significantly increased comparedwith those in control group. Those levels could decreased when concentrationof the lactobacillus Lac 15 was 3×10~6cfu/ml, 3×10~7cfu/ml and 3×10~8cfu/mlcompared with that of HP group.Conclusion:1 The supernate of both the lactobacillus from human and ATCC4356 candirectly repress the growth of Helicobacter pylori.2 Both the lactobacillus from human and ATCC4356 can adhere theSGC7901 cell. All of them can inhibit the adhesive ability to gastric epithelialcell of Helicobacter pylori. And the adhesive ability of the tactobacillusLac15 is the strongest with the same concentration. 3 H.pylori could induce the secretion of IL-8 in the cells whose mechanismsprobably include activating the phosphorylation of p38MAPK.And theLactobacillus from human could inhibit the phosphorylation of p38MAPKand then decrease the secretory volume of IL-8.4 In some concentration range, the lactobacillus from human Lac15 couldinhibit cell inflammatory reaction induced by H.pylori, but if theconcentration was too high they might lose their effects. These maybe explainin partial why lactobacillus as primary common bacterium maintain properproportion concentration is very important for keeping on microecologybalance in stomach.