壳聚糖体内抗幽门螺杆菌作用及其对机体体液免疫反应的调节

The anti H.pylori effect and the regulation effect on humoral ilmnunity by chitosan

目的研究壳聚糖体内抗幽门螺杆菌（Hp）作用，及其对机体体液免疫反应的调节作用。方法建立BALB／c小鼠Hp感染的动物模型后，随机分为8组：（1）对照组；（2）PPI组；（3）AM组；（4）AM＋PPI组；（5）壳聚糖组；（6）壳聚糖＋PPI组；（7）壳聚糖＋AM组；（8）壳聚糖＋AM＋PPI组。分别给予上述药物每日2次灌胃，共2周。停药后4周，处死小鼠，无菌条件下取胃黏膜、唾液和血清。采用定量Hp培养和病理改良Giemsa染色法检测胃黏膜内Hp感染。用ELISA法检测血清、唾液和胃黏膜内Hp抗体，用SP免疫组织化学法检测胃黏膜内分泌型IgA（sIgA）。结果以上8组的却根除率分别为0、0、41．7％、58．3％、58．3％、66．7％、83．3％、91．7％，其中（3）～（8）组的肋根除率与（1）和（2）组比较差异有统计学意义（P〈0．05）。Hp定植密度研究发现各组之间Hp定植密度差异有统计学意义（P〈0．001），坳定植密度在（3）～（8）组显著低于（1）和（2）组（P〈0．05），（7）组显著低于（3）组（P〈0．05），（8）组显著低于（4）组（P〈0．05）。血清中抗Hp IgG、IgG1、IgG2a及唾液中抗Hp IgA含量，各组差异无统计学意义（P〉0．05）。胃黏膜中抗Hp IgA含量，在壳聚糖组和壳聚糖＋AM组显著高于无壳聚糖组（P〈0．05）。胃黏膜sIgA阳性腺体百分率，含壳聚糖组显著高于不含壳聚糖组（P〈0．05）。结论壳聚糖在体内有抗Hp作用，并与AM有协同作用，它与PPI和AM三者联用的Hp根除率高达91．7％，有望成为一抗Hp新药。壳聚糖可促进胃黏膜局部抗Hp IgA和sIgA的产生，因此它在体内的抗Hp作用除了直接杀灭Hp外，其对机体免疫调节效应可能参与了抗菌机制。

Objective To study the anti H. pylorieffect and the regulation on humoral immunity by chitosan. Methods H. pylori infected one-grade female BALB/c mice model were established by inoculating with H. pylori syndey strain 1 and randomly divided into eight groups and administrated by （1） Arabia glue solution （control group）;（2）knsec;（3）Amoxicillin;（4）Amoxicillin plus Losec;（5）chitosan;（6）chitosan pins Lnsec;（7）chitosan plus AmoxiciUin;（8）chitosan plus Amoxicillin plus Losec respectively,twice daily for 14 consecutive days.Four weeks after the last administration, the mice were all killed and gastric mucosa were embedded in paraffin, sectioned and assayed with Giemsa stain. The other gastric mucosa were used to quantitatively culture H.pylori. ELISA was used to detect anti H. pyloriantibodies in serum, sahva and gastric mucosa, SP immuno-histochemical method was used to detect secretory immunoglobulin A （sIgA） in gastric mucosa. Results The eradicate rate was 0, 0, 41.7%, 58.3%, 58.3%, 66.7%, 83.3% and 91.7% respectively, among these eight groups（P〈0.001）;the H. pylori colonized density in control group and Lnsec group was significandy higher than that in other six groups（P〈0.05）;colonizing density in Amoxicillin group was significantly higher than that in chitosan plus Lnsec group, chitosan plus AmoxiciUin group and chitosan plus Amoxicillin plus Lnsec group（P〈0.05）;colonizing density in Amoxicillin plus Losec group was significantly higher than that in chitosan plus Amox icillin plus Lnsec group（ P〈0.05）.There was not difference in the content of anti H.pylori IgG, IgG1, IgG2a in serum and anti H. pylori IgA in saliva among these eight groups（P〉0.05）.The content of anti H. pylori IgA in gastric mucosa in chitosan group and chitosan plus Amoxicillin group was significantly higher than that in those groups without chitosan（P〈0.05）.The labeling index for slgA-positive lumen of glands in groups with chitosan was significantly higher than that in groups without chitosan（P〈 0.05）.Conclusion Chitosan has anti H. pylori effect and synergism with Amoxicilhn in vivo.Chitosan might exert anti H. pylori effect by two mechanisms as follows：chitosan kills H.pylori directly, or immunological regulation effect of chitosan may participate in the anti Hp mechanisms.