摘要
目的研究壳聚糖体内外抗幽门螺杆菌(Hp)作用。方法(1)采用打孔法检测了不同浓度、pH值、脱乙酰度壳聚糖及羧甲基壳聚糖在体外对Hp的抑菌作用。(2)建立BALBc小鼠Hp感染的动物模型后,随机分为8组:①对照组;②PPI组;③AM组;④AM+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进行Hp定量培养和石蜡包埋切片,进行Giemsa染色。结果(1)壳聚糖和羧甲基壳聚糖在体外对3株Hp标准菌株具有普遍的抑菌作用;在pH6～4范围内,随pH值降低,抗菌作用增强,差异有统计学意义(P<0.01),最佳pH值为4;70%、88.5%脱乙酰度壳聚糖及羧甲基壳聚糖的抗Hp作用差异有统计学意义(P<0.05),抑菌强度依次为DD70壳聚糖、DD88.5壳聚糖和羧甲基壳聚糖;在1%～5%浓度范围内羧甲基壳聚糖抗Hp作用差异无统计学意义;在0.5%～2%浓度范围内70%、88.5%脱乙酰度壳聚糖抗Hp作用差异也无统计学意义(P>0.05)。(2)对Hp感染的小鼠,以上8组的Hp清除率分别为0%、0%、41.7%、58.3%、58.3%、66.7%、83.3%、91.7%,其中③～⑧组的Hp清除率显著高于①和②组(P<0.05),⑧组的Hp清除率还显著高于③组(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),Hp定植密度在③～⑧组显著低于①和②组(P<0.05);⑥～⑧组显著低于③组(P<0.05);⑧组显著低于④组(P<0.05)。结论壳聚糖和其衍生物对Hp有普遍的抑菌作用;壳聚糖及其衍生物的抗Hp作用受多种因素影响,其中pH值对壳聚糖抗菌作用的影响最为明显,在pH值6～4范围内,壳聚糖的抗Hp活性随着pH值的下降而显著增强;壳聚糖在体内有抗Hp作用,并与AM有协同作用。
Objective To study the anti-H.pylori effect of chitosan. Methods The effects of three kinds of chitosan—— chitosan with deacetylation degrees（DD） of 70%（DD70）, 88.5 %（DD88.5） and carboxynethyl chitosan——with different concentrations and different pH values on H. pylori standard strains （ 11637, 11639 and SS1） were examined in vitro by bacterium culture. H.pytori infection model was established by inoculating BALB/c mouse with H. pylori SS1. The mice models were randomly divided into eight groups separately injected with ① Arabia glue solution （control group）, ② Losec, ③ Amoxicillin ,④ Amoxicillin plus Losec, ⑤ Chitesan, ⑥ Chitesan plus Losec, ⑦ Chitosen plus Amoxieillin, and ⑧ Chitesan plus Amoxicillin plus Losec. Four weeks after the last administration, the mice were killed and the gastric nmceses were prepared for microscopy and for quantitatively culture of H. pylori. Results （1） Chitesan and carboxynethyl chitesan inhibited all the three H.pylori strains in vitro. In a pH range of 4-6, the lower the pH value, the more significant inhibition of bacteria gnowth （ P 〈0.01）, and the optimal pH value was 4. There were differences in the bacterial, of the three kinds of chitesan （ P 〈 0.05）, which were in ascending order of carboxyuethyl and DD70, DD88.5 chitesan. In the concentration range of 1% - 5 %, carboxynethyl chitosen showed no significant difference in the antibacterial effect; and in the concentration range of 0.5% - 2%, DD70,DD88.5 chitesan showed no significant difference in the antibecterial effect, separately （ P 〉0.05）. （2） There were significant differences in the eradication rates of H.pylori in different groups in vivo（ P 〈 0.001）. The eradication rates of H.pylori in control group and Lesec group were significantly lower than that in the other six groups （ P 〈 0.05） ; the eradication rate in the Amoxicillin group was significantly lower than that in the chitosan plus Amoxicillin plus Losec group（ P 〈 0.05）. There were significant differences in the colonized densities in different groups（ P 〈 0.001 ）. The colonized densities in control group and Losec group were significantly higher than that in the other six groups（ P 〈 0.05）. The colonized density in the Amoxicillin group was significantly higher than that in chitosan plus Losec group, chitosan plus Amoxicillin group and chitosan plus Amoxicillin plus Losec group（ P 〈 0.05）. The colonized density in the Amoxicillin plus Losec group was significantly higher than that in chitosan plus Amoxicillin plus Losec group（ P 〈 0.05）. Conctusion Chitosan and the derivatives have extensive anti-H, pylori activity in vitro, which was influenced by many factors, especially pH value; the anti-bacterial activity of chitosan markedly increased with the decrease of pH value from pH6 to pH4. Chitosan has anti- H. pylori effect in vivo, which was synergistic with Amoxicillin.
出处
《中华微生物学和免疫学杂志》
CAS
CSCD
北大核心
2006年第3期193-198,共6页
Chinese Journal of Microbiology and Immunology
基金
国家自然科学基金资助项目(30060032)
关键词
幽门螺杆菌
壳聚糖
动物模型
抗菌作用
Helicobacter pylori
Chitosan
Animal model
Antibacterial activity
作者简介
通讯作者：谢勇，Emeil：xieyeng-med@tom.com.电话：0791-8692507
