| Objective To explore the effects of the five alkaloids of Sophora Alopecuroides inhibit H.pylori(Helicobacter pylori, H. pylori)in vitro,and to find the active anti-H. pylori components of chinese medicine.Methods Using of improved Colombian culture medium culture H. pylori by exhaust ventilation in vitro isolation,and identified by morphological and biochemical identification of the bacteria to judge.H. pylori strains were frozen using special preservation solution. The selected alkaloids includes Total alkaloids of sophora alopecuroides(TASA), matrine, oxymatrine, sophoridine, sophocarpine. Their antimicrobial effects against H. pylori using the K-B disc agar diffusion methods in a rough range were analysed respectively in the concentration of 200 mg/mL,100 mg/mL,50 mg/mL,25 mg/mL,13 mg/mL, and measure the minimal inhibitory concentration(MIC) by the tube dilution method test.Results 1. Clinical specimens were collected from 197 cases and were cultured 81 strains of isolated H. pylori,the positive rate of 42.12%.2. Solid medium with improved Colombian culture medium culture H. pylori by exhaust ventilation in vitro for 3 to 5 days after the naked eye to the surface of the medium of translucent colonies, colorless, needle size, moist like dew-like. Stained smear microscopy, Gram stain was negative, spiral, curved or gull-like. Urea enzyme test strongly positive oxidase, catalase test was positive. Can be identified by the identification of bacteria isolated from H. pylori.3. H. pylori preservation solution will be paired by the identification of bacteria bacteria, placed Eppendof tube and was preserved under-80℃.4.Take frozen H. pylori strains,37℃water bath for quick thawing,2500r/min centrifugal 5min, discard supernatant, precipitate and mix with a pipette break up, breathe H. pylori solid medium, the third line crossed inoculation method, in micro-aerobic (5% O2,10% CO2 and 85% N2),37℃, humidity> 90% under cultivation 48～72h, the recovery can be successful.5. TASA≥50 mg/mL when inhibition zone, diameter 10 mm; matrine≥100 mg/mL when inhibition zone, diameter 9 mm; Oxymatrine≥100 mg/mL when there inhibition zone, diameter 10 mm; Sophoridine≥200 mg/mL when inhibition zone, diameter 8 mm; sophocarpine concentration in all there was no inhibition zone. Alkaloid that may be in vitro inhibition of H. pylori presence.6.The minimum inhibitory concentration(MIC) of the TASA to the H. pylori was 32 mg/mL and had a certain antimicrobial effect against H. pylori. The MIC of the matrine and the oxymatrine was 64 mg/mL, sophoridine was 128 mg/mL to the H. pylori. Sophocarpine had not bacteriostatic activity to the H. pylori in vitro.Conclusion 1.The solid medium raises H. pylori the optimum condition is: Colombian culture medium 4.25g dissolves in distilled water 200ml, the high pressure antiseptic add-on enters 10% glucose 10ml, antibiotic lml, to escape textile fiber sheep blood 14ml aseptically, micro needs the oxygen (5% O2,10% CO2 and 85% N2), temperature 37℃, humidity>90%.2.Submerged culture H.pylori optimum condition: Cloth bouillon powder 2.8g, dissolves in distilled water 20ml, the high pressure is antiseptic, cools after 50～60℃, adds calf blood serum 2ml,10% glucose 1ml, antibiotic 0.1ml, starch 0.1g, micro needs the oxygen (5% O2,10% CO2 and 85% N2), the temperature 37℃, H.pylori increases the fungus to be rapid. But cannot use in the original generation increasing the fungus raise.3.The preserved fluid low temperature (-80℃) the liquid state saves frozen the strain, the recovery success ratio may reach above 95%.4. Bitter bean total alkali's MIC is 32 mg/mL, has certain inhibitory action in vitro to H.pylori, has the possibility to become substitutes the antibiotic the anti-H.pylori candidate medicine.
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