Shi Li, Hou Xiaohua, Yi Cuiqiong, Zhang Jinkun
Infective Disease Center, Guangzhou 510602.
J Huazhong Univ Sci Technolog Med Sci. 2002;22(2):100-2. doi: 10.1007/BF02857664.
The vacuolated effect of Helicobacter (H. pylori) and its relationship to vacuolated cytotoxin antigen (VacA) were investigated by the method of cytotoxic test and SDS-pobyacrylamide gel electrophoresis (SDS-PAGE). Of the 62 clinical isolates, the broth culture filter (BCF) of 43 strains caused the Vero cell intracytoplasmically vacuolated. H. pylori strains were divided into H. pylori (Toxin+) group with vacuolated effect and H. pylori (Toxin-) group without vacuolated effect. The analysis of the BCF of H. pylori (Toxin+) and that of H. pylori (Toxin-) was studied by SDS-PAGE and Scan reader. A kind of protein with 87 ku molecular weight was recognized in the BCF of 30.23% (13/43) H. pylori (Toxin+) strains but in none of that of H. pylori (Toxin-) strains, the difference was statistically significant (P < 0.05). There was a significant and concordant relationship between OD of the protein band with 87 ku molecular weight and titer of vacuolated activity of H. pylori (Toxin+) (r = 0.67 and P < 0.05 by linear regression analysis). H. pylori strains were divided into H. pylori (Toxin+) group with vacuolated effect and H. pylori (Toxin-) group without vacuolated effect. The vacuolated effect of H. pylori (Toxin+) was caused by the protein with 87 ku molecular weight (VacA).
采用细胞毒性试验及十二烷基硫酸钠-聚丙烯酰胺凝胶电泳(SDS-PAGE)方法,研究幽门螺杆菌(H. pylori)的空泡化效应及其与空泡毒素抗原(VacA)的关系。62株临床分离株中,43株菌的肉汤培养滤液(BCF)可使Vero细胞胞质内出现空泡。幽门螺杆菌菌株分为有空泡化效应的幽门螺杆菌(毒素阳性)组和无空泡化效应的幽门螺杆菌(毒素阴性)组。应用SDS-PAGE及扫描分析仪对幽门螺杆菌(毒素阳性)和幽门螺杆菌(毒素阴性)的BCF进行分析。在30.23%(13/43)的幽门螺杆菌(毒素阳性)菌株的BCF中识别出一种分子量为87 ku的蛋白,而幽门螺杆菌(毒素阴性)菌株的BCF中均未识别出该蛋白,差异有统计学意义(P < 0.05)。分子量为87 ku的蛋白条带光密度值与幽门螺杆菌(毒素阳性)空泡化活性滴度之间存在显著的一致性关系(线性回归分析,r = 0.67,P < 0.05)。幽门螺杆菌菌株分为有空泡化效应的幽门螺杆菌(毒素阳性)组和无空泡化效应的幽门螺杆菌(毒素阴性)组。幽门螺杆菌(毒素阳性)的空泡化效应由分子量为87 ku的蛋白(VacA)引起。