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目的研究重组幽门螺杆菌菌毛蛋白Fla A Ig Y-硫糖铝乙基纤维素微囊对幽门螺杆旋菌(Helicobacter pylori)感染小鼠的治疗和清除效果。方法采用有机溶剂蒸发技术制备经硫糖铝处理后的Ig Y乙基纤维素微囊。模拟胃部环境,检测在不同p H的正常胃蛋白酶浓度(0.02 mg·m L-1)和异常胃蛋白酶浓度(0.04 mg·m L-1)的条件下不同浓度硫糖铝对r Fla A Ig Y的保护效果。将r Fla A Ig Y-硫糖铝微囊按0.8、1.6、3.2及6.4 mg·g-1·d-14种不同剂量灌服实验组小鼠,通过检测治疗前后血清中抗幽门螺杆旋菌抗体水平以及胃黏膜的尿素酶水平,评价不同浓度r Fla A Ig Y对小鼠胃部幽门螺杆旋菌的治疗和清除效果。结果结果表明,40%的硫糖铝处理过的Ig Y在p H 3.0条件下的模拟胃液中能维持77.87%~82.14%的活性达8 h,而在p H 2.0的模拟胃液条件中处理8 h能保持57.76%~63.28%,在p H 1.0模拟胃液中处理8 h能保持21.52%~22.79%。制备的r Fla A Ig Y-硫糖铝乙基纤维素微囊的包封率为83.79%,5 h体外释放率为95.92%。各组小鼠经r Fla A Ig Y微囊治疗后血清中幽门螺杆旋菌抗体水平与阳性对照组相比均显著下降(P<0.001)。当使用剂量为0.8 mg·g-1·d-1时,对幽门螺杆旋菌的清除率为50%,当剂量为1.6 mg·g-1·d-1时,幽门螺杆旋菌清除率为70%,当剂量为3.2 mg·g-1·d-1时,H.pylori清除率为80%,当剂量达到6.4 mg·g-1·d-1时,H.pylori的清除率可达100%。结论 40%的硫糖铝对r Fla A Ig Y具有很好的保护作用,可以作为r Fla A Ig Y的保护剂。r Fla A Ig Y-硫糖铝乙基纤维素微囊具有良好的缓释效果,经口服对小鼠胃部幽门螺杆旋菌感染具有良好的治疗和清除效果。
Objective To investigate the therapeutic and eradication effects of recombinant Helicobacter pylori-infected mice with recombinant Helicobacter pylori micro-vesicles of FlaA Ig Y-sucralfate. Methods Ig Y ethylcellulose microcapsules treated with sucralfate were prepared by organic solvent evaporation technique. The gastric environment was simulated and the effects of different concentrations of sucralfate on the expression of rFlaA at different p H concentrations (0.02 mg · mL -1 -1) and abnormal pepsin concentration (0.04 mg · mL -1) Ig Y protective effect. The r Fla A Ig Y-sucralfate microcapsules were dosed with different doses of 0.8, 1.6, 3.2 and 6.4 mg · g-1 · d-14 to mice in the experimental group. The serum levels of anti-Helicobacter pylori Antibody levels and gastric mucosal urease levels, evaluation of different concentrations of rFla A IgY on gastric Helicobacter pylori treatment and clearance. The results showed that 40% sucralfate-treated Ig Y maintained 77.87% -82.14% activity for 8 h in simulated gastric fluid at p H 3.0 and 8 h in simulated gastric fluid conditions at p H 2.0 h can maintain 57.76% ~ 63.28%, and maintain 21.52% ~ 22.79% in p H 1.0 simulated gastric juice for 8 h. The entrapment efficiency of prepared r Fla A Ig Y-sucralfate microcapsules was 83.79%, and the in vitro release rate was 95.92% in 5 h. Serum Helicobacter pylori antibody levels in each group of mice after treatment with r Fla A Ig Y capsule were significantly lower than those in the positive control group (P <0.001). When the dose was 0.8 mg · g-1 · d-1, the clearance rate of Helicobacter pylori was 50%. When the dose was 1.6 mg · g-1 · d-1, the clearance rate of Helicobacter pylori was 70%. When the dose was 3.2 mg · g-1 · d-1, the clearance rate of H.pylori was 80%. When the dose reached 6.4 mg · g-1 · d-1, the clearance rate of H.pylori reached 100%. Conclusion Forty percent of sucralfate has a good protective effect against rAla A Ig Y and serves as a protective agent for rAla A Ig Y. r Fla A Ig Y-sucralfate microcapsules with good sustained-release effect, oral administration of Helicobacter pylori in mice stomach has a good effect of treatment and clearance.