烯醇化酶在阴道加德纳菌和乳杆菌的表达及其对糖酵解影响的研究
Expression of enolase in Gardnerella vaginalis and vaginal Lactobacillus and its effect on gly-colysis
张瑞 1吴益青 2尹海旭 1李昶 1马爱昕 1王君竹 3白会会 4刘朝晖 4廖秦平5
作者信息
- 1. 北京大学第一医院妇产科,北京 100034
- 2. 西北妇女儿童医院,西安 710061
- 3. 北京大学政府管理学院大数据与公共政策实验室,北京 100091
- 4. 北京妇产医院妇产科,北京 100006
- 5. 北京清华长庚医院妇产科,北京 102218
- 折叠
摘要
目的:研究烯醇化酶在阴道加德纳菌菌株和乳杆菌菌株的胞膜上和胞质内表达的差异情况,并比较两种菌的糖酵解能力的区别.方法:从细菌性阴道病患者阴道分泌物中分离、纯化、鉴定的加德纳菌菌株,从健康女性阴道分泌物中分离乳杆菌菌株,同等条件下,在体外培养一段时间后,检测菌液中残余葡萄糖、乳酸、丙酮酸和三磷酸腺苷(ATP)的浓度.分别提取加德纳菌和乳杆菌的胞膜和胞质蛋白,采用蛋白质免疫印迹法(western blotting)检测胞质和胞膜中的烯醇化酶的情况.结果:加德纳菌的培养基中残余的葡萄糖明显变少,而培养乳杆菌的培养基中残余葡萄糖含量相对较多,两组比较差异有统计学意义(P＜0.05);加德纳菌产生的ATP含量显著高于乳杆菌(P＜0.05).当在反应体系中加入烯醇化酶抗体后,残余葡萄糖的量有所增加,而ATP、乳酸和丙酮酸含量都有所减少;所有加德纳菌和乳杆菌的胞质中均存在烯醇化酶,其中加德纳菌的烯醇化酶的含量显著高于乳杆菌(P＜0.05).仅部分加德纳菌菌株和乳杆菌菌株的胞膜上存在烯醇化酶,加德纳菌胞膜上的烯醇化酶的含量显著高于乳杆菌(P＜0.05).结论:烯醇化酶在阴道加德纳菌和乳杆菌的表达存在一定差别,这种差别一定程度上影响了两种菌种代谢的能力.
Abstract
Objective:To investigate the differential expression of enolase in the membrane and cytoplasm of Gardnerella vaginalis and Lactobacillus strains,and to compare the differences in their glycolytic abilities.Meth-ods:Gardnerella strains were isolated,purified,and identified from vaginal secretions of patients with bacterial vaginosis.Lactobacillus strains were isolated from vaginal secretions of healthy women and cultured in vitrofor a period of time under the same conditions.The residual concentrations of glucose,lactate,pyruvate,and adenos-ine triphosphate(ATP)in the bacterial solution were detected.The membrane and cytoplasmic proteins of Gard-nerella and Lactobacillus were extracted respectively,and the enolase levels in the cytoplasm and membrane were detected by western blotting.Results:The residual glucose content in the culture medium of Gardnerella was decreased significantly,while the residual glucose content in the culture medium of Lactobacillus was rela-tively higher,and there was significant difference between the two groups(P＜0.05);the amount of ATP pro-duced by Gardnerella was significantly higher than that of Lactobacillus(P＜0.05).When enolase antibodies were added to the reaction system,the amount of residual glucose was increased,while the amount of ATP,lac-tate,and pyruvate was decreased.Enolase was present in the cytoplasm of all Gardnerella and Lactobacillus,and the content of enolase in Gardnerella was significantly higher than that in Lactobacillus(P＜0.05).Only some Gardnerella and Lactobacillus strains had enolase on their cell membranes,and the content of enolase on Gard-nerella cell membranes was significantly higher than that on Lactobacillus cell membranes(P＜0.05).Conclu-sion:There is a certain difference in the expression of enolase between Gardnerella vaginalis and Lactobacillus,which to some extent affects the metabolic ability of the two strains.
关键词
细菌性阴道病/烯醇化酶/阴道加德纳菌/阴道乳杆菌/糖酵解Key words
bacterial vaginosis/enolase/Gardnerella vaginalis/vaginal Lactobacillus/glycolysis引用本文复制引用
基金项目
国家自然科学基金资助项目(81471420)
教育部博士点新教师基金项目(20130001120051)
出版年
2024
国家科技期刊平台
NSTL
万方数据