| Aims:To extract garlic oil and observe its stability in ampoule dosage form;To study the inhibition effect of garlic oil on helicobacter pylori in Vitro and Vivo;To probe a well new anti-H.pylori drug from Alliun sativum L.Methods: (1) Extracted garlic oil with organic solvent and repeat once again with different solvent ; Identified garlic oil and quantitatively analyzed its antibacterial compound Diallyl disulfide and Diallyl trisulfide by GC/MS ;0bserve its stability made into ampoule dosage form and reserved lower temperature by GC. (2) In Vitro experiment , the multiple proportion dilution of garlic oil were select to observe the minimum inhibitiory concentration (MIC)against NCTC11639 and Yunnan H. pylori; (3) Amplify the vacA gene subtypes and cagA genes 29 H. pylori strains using polymerase chain reaction(PCR);Isolated, identified, transferred and reserved the Yunnan H. pylori mouse-passaged strainscarrying cagA genes and vacA gene; The animal model of infection is established by H. pylori mouse-passaged strains infected; Treated BALB/c mice infected H. pylori with Garlic oil and to observe the result and untoward reaction.Results: (1) The extracted oil rate of purple clove of singular garlic in Kunming was 0. 132%; Diallyl disulf ide , Diallyl trisulfide extracted from garlic oil were 13. 5%, 14.8% respectively; In three months, Diallyl disulfide were 13. 4743%, 13. 3603%, 13.8852%, Diallyl trisulfide were 14.8408%, 14. 9045% , 14.2677%, respectively .(2) 100% H.pylori strains inhibited when the concentration is 165 μg/ml, 83.3% H.pylori strains inhibited when the concentration is 82. 5 u g/ml, 26.7% H. pylori strains inhibited when the concentration is 41. 25 μ g/ml. Complete lack of resistrance of bacteria to garlic oil has been found. (3) CagA is detected in 86.2%(25/29) in the 29 Yunnan isolates, vacA subtype sl/m2, s1/m1b, s2/m2, sl/mlb+m2and s1/m are detected in 55.2% (16/29), 13.8% (4/29), 3.5% (1/29), 17.2% (5/29) and 10.3% (3/29) respectively;(4) BALB/c mice were inoculated orally through a feeding tube with 4 isolates of H.pylori carrying cagA genes and vacA gene successfully . The infected rate of H.pylori was 33.3% (3/9) ;It is unsuccessful with NCTC11639, There was significantdifference between the infected rate of H. pylori with Yunnan strains and NCTC11639 strains (P<0.05) . Isolated Helicobacter pylori strains from mice gastric mucus and transferred three times, then reserved. It is inoculated successfully the H. pylori mouse-passaged strains. Amplified the vacA gene subtypes and cagA genes of the H. pylori mouse-passaged strains using polymerase chain reaction(PCR), it is confirmed that the stain is the same before inoculated and after. (5)After inoculated with the H. pylori mouse-passaged strains, at 4 weeks , H. pylori infection was assessed by bacterial culture, urease assay and histological method, the infection rate of infection group was 100% (10/10) . No H, pylori were found in normal control group (0/10) . There was significant difference between infection group and normal control group(P<0. 05) . (6) The mice were sacrificed 1 day after the cessation of treatment , the clearance rate of H. pylori in garlic oil treated group is 60%(6/10),The mice were sacrificed 4 weeks after the cessation of treatment , the eradication rate of H.pylori in garlic oil treated group is 70%(7/10), the combined eradication rate of H.pylori is 65%(13/20). During the course of treatment, only a mice was found anorexia, after treatment , it recovered quickly. No H. pylori were found in normal control mice(0/20) . H. pylori were found in untreated infectioncontrol mice and the control infection mice treated with cooking oil (40/40) . There was significant difference between garlic oil treated group and the rest three groups(P<0.001) .Conclusions:(1) The extracted oil rate of purple clove of singular garlic in Kunming was 0.132%. Antibacterial compound in garlic oil is high,Diallyl disulfide , Diallyl trisulfide extracted from garlic oil were 13. 5%, 14.8... |
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