Study On Expression Of AphA And AhpC Gene Of Helicobacter Pylori Under The Pressure Of Medicine: Clarithromycin Medical

Objective:The standard strain of Helicobacter pylori was cultured.The expression of two genes of Helicobacter pylori was analyzed in order to study growth state of Helicobacter pylori under the pressure of medicine:clarithromycin.The change of expression of aphA and ahpC gene was investigated during the course of growth of Helicobacter pylori under clarithromycin.Methods:The number standard strain of Helicobacter pylori was 26695(NCTC).Helicobacter pylori were cultured on fresh 7%-10%Helicobacter pylori selective medium,and were cultured under microaerobic conditions at 37℃for 3-5 days. The two different groups were divided,one group was that Helicobacter pylori was induced by clarithromycin,and the other group was not induced by clarithromycin.Then Helicobacter pylori was cultured in 10 days for each group. The E-Test was used to determine the resistance and susceptibility of Helicobacter pylori strains to clarithromycin. Furthermore,the concentration of medicine clarithromycin was less than 0.016μg/mL.The Helicobacter pylori was cultured in 10 days under the above concentration of clarithromycin.The total RNA was extracted,and then the cDNA of Helicobacter pylori was synthesized through reverse transcription reaction.Therefore,the specific primer pairs for Helicobacter pylori were used to amplify the gene aphA and ahpC,and then the gyr house keeping gene was chosen as control.Finally,the expression changes of aphA and ahpC were analyzed through agarose gel electrophoresis.Results:Helicobacter pylori was induced by using the concentration 0.016μg/mL clarithromycin.Then the expression changes of aphA and ahpC were analyzed through agarose gel electrophoresis.The results were compared with the results of agarose gel electrophoresis from gyr gene.The above results were analyzed by using the Bio-rad soft.After 10 culture days,the results of analysis showed that the ratio of aphA gene and gyr gene from two groups,one group was that Helicobacter pylori was induced by clarithromycin,and the other group was not induced by clarithromycin,were 0.63, 0.68,1.16,0.70,1.16,1.03,1.20,1.04,1.54,1.41,0.45, 0.77,0.74,0.75,0.77,0.64,0.70,1.02,0.85,0.73 respectively.Meanwhile,the ratio of ahpC gene and gyr gene were 1.67,1.32,1.35,1.56,1.23,1.14,1.52,1.46,1.25,1.17, 1.22,1.27,1.39,1.22,1.46 respectively.Then the ratio of aphA gene from two groups were 1.4,0.88,1.56,0.93,1.50, 1.60,1.71,1.01,1.81,1.93;the ratio of ahpC gene from two groups were 1.36,1.03,0.97,1.32,1.15,0.87.Conclusions:Through induction by clarithromycin,the expression of aphA and ahpC genes of bacterial strain had been increased obviously.The above results indicated that the expression of aphA and ahpC gene of Helicobacter pylori may be changed under the pressure of medicine:clarithromycin.The result of this paper provided a reliable experiment data for Helicobacter pylori in order to further study the pathway of gene signal regulation on the gene level under the pressure from other enviromental factors.