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Background and Purpose: Helicobacter pylori (Helicobacter pylori, Hp) can cause chronic gastritis, gastric ulcer, duodenal ulcer, upper gastrointestinal diseases related to gastric cancer, gastric mucosa-associated lymphoid tissue lymphoma. This stage clinical treatment of Hp infection is still used antibiotics, but such treatment to poor patients obedient, expensive, strong side effects and the growing problem of bacterial resistance to antibiotics and other problems. Natural plant ingredients to avoid these drawbacks, there are study found that some natural plant ingredients Hp show some antimicrobial effects, including natural plant ingredients mastic gum, garlic oil, berberine. This study selected the frankincense gum and recognized broader antibacterial spectrum garlic oil, berberine extract and berberine as object of study, these different kinds of natural plant ingredients anti Hp activity conducted a preliminary study, through in vitro and in vivo tests proved its in vitro the efficacy of antibacterial activity and Clear Hp resistance induced reaction experimental research and toxicology studies to investigate the resistance and security through pharmacokinetic study to determine the pharmacokinetic parameters for the development and evaluation of drug theoretical basis. Method: a natural plant ingredients vitro antibacterial activity. Double dilution agar plate method for the determination of the minimum inhibitory concentration (MIC); tube double dilution method for the determination of the minimum bactericidal concentration (MBC); colony counting method to draw the killing curve; explore the effects of inoculum size and medium pH on drug measured the MIC ; scanning and transmission electron microscopy to observe changes the bacterial morphology before and after the drug action. Natural plant ingredients vivo anti-Hp activity tests. (1) the therapeutic dose of the mastic gum screening test. Gerbils infected with 180 divided into the frankincense gum drug dose group (15 mg, 7.5 mg, 3.75mg), solvent treatment group, no drug treatment group and the treatment groups (omeprazole 3mg and clarithromycin 2mg), respectively. continuous medication treatment of 14d. Medicate 8d, after stopping the first day after stopping 20d slaughter gerbils stomachs organizations do urease test and histopathological examination, isolation and culture, PCR assay. Clearance rate was observed. (2) a variety of natural plant ingredients clearance test. Gerbils infected with divided into 15mg frankincense gum group 140, 15mg Garlic oil group, 15mg the Huanglian extract group, 15mg berberine group, the solvent treatment group, untreated treatment group and positive drug treatment group (Omeprazole 3mg and clarithromycin 2mg), infected gerbils continuous medication therapy 14d Hp whole cell vaccine 0,1,2,3 w intramuscular immunization of gerbils with 15mg frankincense gum treatment, after stopping 1d, stop the end of the 20d or immune drug Rule 14d slaughter gerbils, whichever gastric tissue do urease test and histopathological examination, isolation and culture, PCR detection. Clearance rate was observed. 3.Hp vitro resistance induction experiments. Multi-step induction method, the choice of a very high rate of clinical resistance to antibiotics metronidazole and anti-Hp good efficacy of antibiotic clarithromycin as control. Frankincense gum, garlic oil, berberine extract, berberine, metronidazole and clarithromycin increasing concentrations in the medium turn species Hp, observed changes in the MIC. Decreased sensitivity to induced strain on the medium without drugs transferred species is detected after the MIC value. Determination of frankincense gum, frankincense gum, garlic oil, berberine extract and berberine metronidazole, clarithromycin MIC values ??decreased susceptibility to induced strain. 4. The detected antibacterial effect of law indicators frankincense gum pharmacokinetic parameters. By acute toxicity tests and long-term toxicity tests for toxicological studies frankincense gum. Maximum tolerated dose, fed mice to observe the death of mice within 7d; daily orally at different doses in rats 1 for 30 consecutive days to detect blood biochemical analysis, organ weight and weight detection , organ tissue to do a biopsy. Results: four natural plant ingredients showed in vitro antibacterial activity. Frankincense gum, garlic oil, berberine extract and berberine respectively MIC values ??of HpSS1, 125ug/ml 82.5ug/ml 1000ug/ml and 500ug/ml; MIC values ??of Staphylococcus aureus ATCC25923 were 1000ug/ml The the, 82.5ug/ml, 4000ug/ml and 2000ug/ml; E. coli ATCC25922 MIC values ??were 2000ug/ml, 330ug/ml, 8000ug/ml and 2000ug/ml. The the MBC of their HpSS1 to 500ug/ml 165ug/ml 1000ug/ml and 500ug/ml; Staphylococcus aureus ATCC25923 MBC were 2000ug/ml, 330ug/ml, 8000ug/ml and 4000ug/ml,; the large intestine The bacilli ATCC25922 MBC were 8000ug/ml, 1320ug/ml, 32000ug/ml and 8000ug/ml. From the kill curve, the three drugs in concentration 1/2MIC and 1/4MIC, three strains than those in the control strain slow-enrichment. When concentration to 2MIC and 4MIC, Hp, Staphylococcus aureus and Escherichia coli showed strong bactericidal effect, there is a clear dose-response relationship. The frankincense gum vitro antibacterial activity by the amount of inoculum medium pH value. When the inoculation amount was increased to 10 9 CFU / ml, the medium pH is reduced to 4.0 or an increase of 9.0 hours, the weakening of the antimicrobial activity of mastic gum. The frankincense gum role Hp, electron microscope, some body smaller, sparse cytoplasm individual cell wall damage, cytoplasmic spillover. Natural plant ingredients vivo anti-Hp activity tests. Frankincense gum treatment (1) Section 8d the 3.75mg dose group cleared 25% 7.5mg dose group cleared rate is 0%, the 15mg dose group clearance rate is 50%, after stopping 1d, the three dose groups The clearance rates are 100%. After stopping 20d 3.75mg dose group clearance rate is 25%, 7.5mg dose group Clear the rate is 50%, and the 15mg dose group clearance rate is 100%. Positive drug treatment group were 100% clearance rate of the three time periods. (2) the application of natural plant ingredients frankincense gum, garlic oil, berberine extract, berberine and Hp whole bacteria vaccine, frankincense gum Hp whole cell vaccine to clear the treatment of infected gerbils after treatment 1d after the first four drugs The clearance rate was 90%, 70%, 70% and 60%, respectively. 20d after treatment, they are cleared of infected gerbils was 100%, 80%, 70%, 70%, 70% and 100%, 100%, the removal rates of the two time periods of the positive drug treatment group. 3.Hp vitro resistance induction experiments. Frankincense gum, garlic oil, berberine extract and berberine group MIC values ??remain unchanged, and that is not induced in the process of in vitro resistance induced culture of frankincense gum, garlic oil, berberine extract and berberine resistance Hp strains, the strain could grow in the medium containing 64MIC metronidazole. In the training process, the sensitivity of the strains to metronidazole decreased gradually increased, MIC value last reached 64 μg · mL -1 . Metronidazole-resistant strains (MR) was transferred to plant 10 times in drug-free medium MR ', the MIC values ??unchanged. Frankincense gum, garlic oil, berberine extract and berberine did not affect the MIC of MR. 4 the frankincense gum pharmacokinetic parameters for: Ka-= 0.666 h -1 of Ke = 0.143 h -1 t 1/2 (Ka) = 1.041 h, t 1/2 (Ke) = 4.856h, Lagtime = 0.574h, T (peak) = 3.0h, C (max) = 0.9503ug/ml, AUC = 3.2605ug/ml · h, CL / F = 38.03g/kg · h / (ug / ml), V / F = 266.8 (g / kg) / (ug / ml). 5. Mice 7d within no death, the general condition of the rats, blood, blood biochemical indicators, organ weight and weight index detected no abnormal biopsy no abnormal changes. Conclusion: 1. Frankincense gum, garlic oil, the Huanglian extract and berberine have anti-Helicobacter pylori, Staphylococcus aureus and Escherichia coli activity. Frankincense gum effective in the treatment of Hp infection in vivo experimental low-dose group, but after stopping recurrence. 15mg dose group clearance rate was 100%, and there is no recurrence. 15mg the frankincense gum with the same dose of garlic oil, berberine extract, berberine and Hp bacteria vaccine compared with in vivo tests, they are effective, frankincense gum group effect. The frankincense gum and Hp whole cell vaccine common use, did not produce a mutually antagonistic role. 3.Hp on frankincense gum, garlic oil, berberine extract and berberine is not easy to produce drug resistance, Metronidazole induced a highly resistant strain, metronidazole-resistant strains of frankincense gum the extract of garlic oil, berberine and berberine remain sensitive, frankincense gum, garlic oil, berberine extract and berberine have good development prospects of the natural anti-Hp infection drugs. 4 The the frankincense gum pharmacokinetic one-compartment open model. The drug is administered orally, more rapid absorption, while eliminating the relatively slow. Frankincense gum by the acute toxicity test rats with long-term toxicity tests showed no apparent toxic effects, to be used with a dose of security.
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