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目的了解浙江地区消化性溃疡(PU)和慢性胃炎(CG)患者感染幽门螺杆菌(Hp)vacA基因的变异及其空泡毒素表达情况。方法从70株Hp中选择部分菌株测定其vacA基因的s区和m区核苷酸序列。用细胞培养的方法检测Hp菌株的空泡毒性。结果6株s1a型Hp菌株的s区扩增产物与报道的s1a型60190株核苷酸序列相似性为93.2%~98.3%,4株m2型Hp菌株的m区扩增产物与报道的m2型87-203株核苷酸序列之间相似性为93.8%~97.6%。1株m1b型Hp菌株的m区扩增产物与报道的m1型60190株相应序列相似性为87.3%,与m2型87-203株相应序列之间相似性为71.7%。所有5株s1a/m1型Hp对HeLa、RK-13和SGC-7901三种细胞均有明显的空泡毒作用;仅27.9%(12/43)的s1a/m2型菌株对HeLa细胞表现为空泡毒作用,但分别有65.1%(28/43)和62.8%(27/43)的菌株对RK-13细胞和SGC-7901细胞有空泡毒作用。所有s1a/m1b和s1a/m1b-m2型Hp菌株对三种细胞均有明显的空泡毒作用,以RK-13细胞计,81.0%(17/21)的s1a/m1b型Hp菌株和1株s1a/m1b-m2型有空泡毒性。PU组感染的Hp菌株空泡毒性(84.4%)强于CG组(60.5%),差异有统计学意义(P<0.05)。结论浙江地区患者感染的Hp菌株vacA基因变异主要在于m区。vacA基因空泡毒性的表达在HeLa细胞中低毒性的s1a/m2型菌株在RK-13和SGC-7901细胞中仍有较高的空泡毒性。m1b型Hp空泡毒性强度介于m1和m2型之间。PU患者感染的Hp菌株其空泡毒性明显强于CG组。
Objective To investigate the variation of vacA gene in Helicobacter pylori (Hp) and the expression of vacuolating toxin in patients with peptic ulcer (PU) and chronic gastritis (CG) in Zhejiang province. Methods A total of 70 strains of Hp were selected to determine the nucleotide sequences of the sAs and m regions of the vacA gene. Vacuolar toxicity of Hp strains was tested by cell culture. Results The similarity of the amplified region of s1a of 6 s1a strains to the reported s1a 60190 was 93.2% ~ 98.3%. The amplified products of m region of 4 strains of m2 type Hp were similar to those reported in m2 The similarity between 87-203 nucleotide sequences was 93.8% ~ 97.6%. The similarity between the amplified products of m region of m1b strain m1b and that of the reported m1 type 60190 strain was 87.3%, which was 71.7% compared with the corresponding sequence of m2 type 87-203 strain. All 5 strains of s1a / m1 Hp had obvious cytotoxicity to HeLa, RK-13 and SGC-7901 cells, while only 27.9% (12/43) of s1a / m2 strains showed empty HeLa cells However, 65.1% (28/43) and 62.8% (27/43) of strains had cytotoxicity to RK-13 cells and SGC-7901 cells, respectively. All s1a / m1b and s1a / m1b-m2 Hp strains had obvious vacuolar toxicity to all three kinds of cells. 81.1% (17/21) s1a / m1b type Hp strains and 1 strain s1a / m1b-m2 type vacuolar toxicity. Hypoxic virulence (84.4%) in PU group was stronger than that in CG group (60.5%), the difference was statistically significant (P <0.05). Conclusion The variation of vacA gene in Hp isolates from patients in Zhejiang province mainly lies in m area. VacA gene vacuole toxicity expression The low-toxicity type s1a / m2 strains in HeLa cells still have higher vacuolar toxicity in RK-13 and SGC-7901 cells. The toxicity intensity of m1b-type Hp vacuole is between m1 and m2. Hp strains infected with PU had significantly higher vacuolization toxicity than CG group.