中华微生物学和免疫学杂志
中華微生物學和免疫學雜誌
중화미생물학화면역학잡지
CHINESE JOURNAL OF MICROBIOLOGY AND IMMUNOLOGY
2010年
2期
120-125
,共6页
顾迟%周宏伟%徐翔%潘文胜%张嵘%陈功祥
顧遲%週宏偉%徐翔%潘文勝%張嶸%陳功祥
고지%주굉위%서상%반문성%장영%진공상
尿道致病性大肠杆菌%usp基因%HeLa细胞%快速早期凋亡
尿道緻病性大腸桿菌%usp基因%HeLa細胞%快速早期凋亡
뇨도치병성대장간균%usp기인%HeLa세포%쾌속조기조망
Uropathogenic Escherichia coli%usp gene%HeLa cell%Rapid early stage apoptosis
目的 了解致病基因在尿液分离大肠杆菌中的分布情况,分析由携带usp致病基因的尿道致病性大肠杆菌介导的HeLa细胞快速凋亡.方法 采用PCR检测6种尿道致病性大肠杆菌相关致病基因在28株尿液分离的大肠杆菌的分布;通过黏附试验和锥虫蓝染色对尿液分离的大肠杆菌进行初步致病表型筛选;采用Annexin V/PI法进一步检测大肠杆菌引起HeLa细胞早期凋亡的能力;电子显微镜观察细胞凋亡特征.结果 在28株尿液分离的大肠杆菌中,检测到usp基因阳性6株.通过致病表型筛选试验筛选到2株强致病性大肠杆菌(6N和27N),其中6N菌株携带usp基因,而27N除了没有检测到usp基因,其他致病基因与6N相同.它们可以在4 h破坏大量HeLa细胞,使HeLa死亡;流式细胞仪检测凋亡结果 显示,6N菌株在1.5 h可以诱导20.75%的HeLa细胞发生早期凋亡,而27N只有1.55%的HeLa细胞发生早期凋亡.电镜结果 从形态学证实6N菌株可以快速引起HeLa细胞发生早期凋亡.结论 携带usp基因的尿道致病性大肠杆菌可以诱导HeLa发生快速早期凋亡.
目的 瞭解緻病基因在尿液分離大腸桿菌中的分佈情況,分析由攜帶usp緻病基因的尿道緻病性大腸桿菌介導的HeLa細胞快速凋亡.方法 採用PCR檢測6種尿道緻病性大腸桿菌相關緻病基因在28株尿液分離的大腸桿菌的分佈;通過黏附試驗和錐蟲藍染色對尿液分離的大腸桿菌進行初步緻病錶型篩選;採用Annexin V/PI法進一步檢測大腸桿菌引起HeLa細胞早期凋亡的能力;電子顯微鏡觀察細胞凋亡特徵.結果 在28株尿液分離的大腸桿菌中,檢測到usp基因暘性6株.通過緻病錶型篩選試驗篩選到2株彊緻病性大腸桿菌(6N和27N),其中6N菌株攜帶usp基因,而27N除瞭沒有檢測到usp基因,其他緻病基因與6N相同.它們可以在4 h破壞大量HeLa細胞,使HeLa死亡;流式細胞儀檢測凋亡結果 顯示,6N菌株在1.5 h可以誘導20.75%的HeLa細胞髮生早期凋亡,而27N隻有1.55%的HeLa細胞髮生早期凋亡.電鏡結果 從形態學證實6N菌株可以快速引起HeLa細胞髮生早期凋亡.結論 攜帶usp基因的尿道緻病性大腸桿菌可以誘導HeLa髮生快速早期凋亡.
목적 료해치병기인재뇨액분리대장간균중적분포정황,분석유휴대usp치병기인적뇨도치병성대장간균개도적HeLa세포쾌속조망.방법 채용PCR검측6충뇨도치병성대장간균상관치병기인재28주뇨액분리적대장간균적분포;통과점부시험화추충람염색대뇨액분리적대장간균진행초보치병표형사선;채용Annexin V/PI법진일보검측대장간균인기HeLa세포조기조망적능력;전자현미경관찰세포조망특정.결과 재28주뇨액분리적대장간균중,검측도usp기인양성6주.통과치병표형사선시험사선도2주강치병성대장간균(6N화27N),기중6N균주휴대usp기인,이27N제료몰유검측도usp기인,기타치병기인여6N상동.타문가이재4 h파배대량HeLa세포,사HeLa사망;류식세포의검측조망결과 현시,6N균주재1.5 h가이유도20.75%적HeLa세포발생조기조망,이27N지유1.55%적HeLa세포발생조기조망.전경결과 종형태학증실6N균주가이쾌속인기HeLa세포발생조기조망.결론 휴대usp기인적뇨도치병성대장간균가이유도HeLa발생쾌속조기조망.
Objective To understand the distribution of pathogenic genes in uropathogenic Esche-richia coli (UPEC) from urinary specimen and to analyze the pathogenicity of UPEC and their mechanism of apoptosis to HeLa cells. Methods We have analyzed 6 pathogenic genes among the 28 strains of the clini-cally isolated E. coli from urinary tract infected patients. The 6 pathogenic genes were surveyed by using the PCR amplification of the target genes. The adhesion experiments and tryphan-blue staining was used to screen the phenotype of the pathogenic strains, while Annexin V/PI method was applied to study the strains to cause apoptosis of the HeLa cells, which was further confirmed with electronic microscopy. Results We have detected 6 strains that carried the usp gene. Phenotype screening identified two high virulent isolates (strain 6N and 27N) from the 6 strains. Strain 27N and 6N contained very similar virulence gene profile ex-cept that strain 27N did not contain usp gene. Both strains can destroy HeLa cell within 3 hours causing cell death. Results of apoptosis detected by flow cytometry revealed that strain 6N induced 20.75% of HeLa cells to an early stage apoptosis within 1.5 hours. On the other hand, strain 27N induced only 1.55% of HeLa cells to apeptosis. Conclusion High virulent UPEC strain carrying usp gene can induce HeLa cell rapid early apoptosis.
