The Study Of Allicin Decreasing The Inhibition Of Helicobacter Pylori On Gap Junction Intercellular Communication Of Gastric Epithelial Cells

Objective To observe the effects of Helicobacter pylori and allicin intervention on the expression of connexin37,connexin43 and gap junction intercellular communication of gastric epithelial cells MGC-803 in vitro,identifying allicin can decrease the inhibition of H.pylori on gap junction intercellular communication of gastric epithelial cells.Methods Human gastric epithelial cell line MGC-803 cell was cultured when adhering to wall after a night in vitro,the experiment was divided into some groups:①control groups:cells were non-cultured with H.pylori or allicin for 24,48 and 72 hours;②allicin groups:cells were co-cultured with allicin at concentrations of 3μg/ml,6μg/ml,9μg/ml,12μg/ml for 24,48 and 72 hours respectively;③H.pylori groups:cells were co-cultured with CagA~+ and VacA~+ international standard Helicobacter pylori line J99 at bacterium/cells ratio of 100:1 for 24 hours;④allicin plus H.pylori groups:cells were co-cultured with allicin at a concentration of 12μg/ml for 24 hours before co-cultured with NCTC J99 for 24 hours;⑤H.pylori plus allicin groups:cells were co-cultured with NCTC J99 for 24 hours before co-cultured with allicin at a concentration of 12μg/ml for 24 hours.RT-PCR method was applied to detect the expression of connexin37,connexin43mRNA in cells.Scrape-loading dye transfer method Was applied to detect gap junction intercellular communication.To use paper diffusion method to observe the diameter of the inhibition rings of allicin to H.pylori.The cell proliferation was examined by methyl tetrazolium(MTT)method.Results 1.Control groups:MGC-803 cell line expressed Cx37mRNA, there was no Cx43mRNA expression,cell gap junction intercellular communication was weak.2.Allicin groups:The expression of Cx37mRNA was higher than the control groups(p＜0.01)and up-regulated with the increasement of concentration and action time of allicin.There was significant difference in expression of Cx37mRNA between the allicin groups(P＜0.05).There was no Cx43 mRNA expression,gap junction intercellular communication was improved.3. H.pylori groups:The expression of Cx37mRNA was lower than the control group(p＜0.01).There was no Cx43mRNA expression.4.The allicin plus H.pylori groups and the H.pylori plus allicin groups:There were no statistics differences in expression of Cx37 between the two groups(p＞0.05),the expression of Cx37mRNA were higher than the H.pylori groups(p＜0.01),and cell gap junction intercellular communication was improved.5.The average diameter of the inhibition rings of allicin to H.pylori were 13±3.49mm,more than 7mm.6.Allicin suppressed the cell proliferation at a dose and time-dependent manner.Conclusions1.H.pylori can down-regulate expression of Cx37mRNA in MGC-803 cell at the transcription level.2.Allicin can up-regulate expression of Cx37mRNA in MGC-803 cell at the transcription level in relation to the concentration and time action of allicin,improve GJIC function.3 Allicin can sterilize H.pylori,suppress the MGC-803 cell proliferation,up-regulate expression of Cx37 expression and decrease the inhibition of H.pylori on gap junction intercellular communication of gastric epithelial cells.