摘要
目的建立幽门螺杆菌(Hpylori,Hp)体外黏附MKN45细胞模型,为抗黏附药物的筛选和评价奠定基础。方法采用FITC荧光素标记Hp,以FITC-Hp菌液荧光密度为指标,分别评价FITC用量、FITC标记时间等关键因素,优化FITC标记Hp最佳条件;同时用细胞爬片-革兰染色法确定Hp黏附细胞的最佳时间。结果终浓度为5mg·mL-1的FITC和Hp菌液(浓度为1×106cuf.ml-1)共同孵育1h,为FITC标记Hp的最适条件;Hp黏附MKN45细胞的最佳孵育时间为1h。通过该模型测定芦荟粗多糖抗黏附活性,显示芦荟粗多糖有效抑制Hp黏附MKN45细胞的浓度分别为1g.L-1(P<0.05)以及2g.L-1(P<0.01),并呈剂量依赖性。结论FITC荧光标记检测Hp黏附MKN45细胞模型建立并可用于检测化合物抗Hp黏附活性研究。
Aim To establish the adherence of H pylori to MKN45 cell model for screening and evaluating anti-adhesive drugs.Methods H pylori suspension was incubated with FITC solution in different conditions（FITC concentration and incubation time）to optimize the labeling conditions.At the same time,Gram staining was used to determine the optimal adhesion time of H pylori to MKN45 cells.Results H pylori suspension（1×10^6 cuf·ml^-1）incubating with FITC（5 mg·L^-1）for 1 h could achieve the optimal effect for FITC-labeling H pylori.Besides,Gram staining indicated that the optimal adhesion time for H pylori to MKN45 cells was 1 h.Based on the model,the crude Aloe polysaccharide was evaluated for its anti-adhesive activity and the results showed it could significantly reduce the adherence of H pylori to MKN45 cells at the concentration of 1 g·L^-1（P〈0.05）and 2 g·L^-1（P〈0.01）in a dose dependent manner.Conclusion The adherence of H pylori to MKN45 cells model has been established in this study and can be used for the evaluation of anti-adherent drugs.
出处
《中国药理学通报》
CAS
CSCD
北大核心
2010年第5期683-686,共4页
Chinese Pharmacological Bulletin
基金
国家自然科学基金资助项目(No30600117)
中国科学院上海有机化学研究所生命有机化学国家重点实验室开放课题
南京大学医药生物技术国家重点实验室开放课题
