Luo Yi-Hui, Yan Jie, Mao Ya-Fei
Department of Medical Microbiology and Parasitology, College of Medical Science, Zhejiang University, 353 Yan An Road, Hangzhou 310031, Zhejiang Province, China.
World J Gastroenterol. 2004 Jul 15;10(14):2055-9. doi: 10.3748/wjg.v10.i14.2055.
To determine the biological activity of Helicobacter pylori (H. pylori) lipopolysaccharide (H-LPS) and understand pathological correlation between H-LPS and human chronic gastritis and peptic ulcer.
H-LPS of a clinical H. pylori strain and LPS of Escherichia coli strain O55:B5 (E-LPS) were extracted by phenol-water method. Biological activities of H-LPS and E-LPS were detected by limulus lysate assay, pyrogen assay, blood pressure test and PBMC induction test in rabbits, cytotoxicity test in NIH 3T3 fibroblast cells and lethality test in NIH mice. By using self-prepared rabbit anti-H-LPS serum as the first antibody and commercial HRP-labeled sheep anti-rabbit sera as the second antibody, H-LPS in biopsy specimens from 126 patients with chronic gastritis (68 cases) or gastric ulcer (58 cases) were examined by immunohistochemistry.
Fibroblast cytotoxicity and mouse lethality of H-LPS were weaker than those of E-LPS. But the ability of coagulating limulus lysate of the two LPSs was similar (+/0.5 ng/mL). At 0.5 h after H-LPS injection, the blood pressures of the 3 rabbits rapidly declined. At 1.0 h after H-LPS injection, the blood pressures in 2 of the 3 rabbits fell to zero causing death of the 2 animals. For the other one rabbit in the same group, its blood pressure gradually elevated. At 0.5 h after E-LPS injection, the blood pressures of the three rabbits also quickly declined and then maintained at low level for approximately 1.0 h. At 0.5 h after injection with H-LPS or E-LPS, PBMC numbers of the rabbits showed a remarkable increase. The total positivity rate of H-LPS from 126 biopsy specimens was 60.3%(76/126). H-LPS positivity rate in the biopsy specimens from chronic gastritis (50/68, 73.5%) was significantly higher than that from gastric ulcer (26/58, 44.8%) (chi(2)=10.77, P<0.01). H-LPS positivity rates in biopsy specimens from chronic superficial gastritis (38/48, 79.2%) and chronic active gastritis (9/10, 90.0%) were significantly higher than that of the patients with atrophic gastritis (3/10, 30.0%) (chi(2)=7.50-9.66, P<0.01).
The biological activities of H-LPS were weaker than those of E-LPS, the activities of H-LPS of lowering rabbit blood pressure and inducing rabbit PBMC were relatively stronger. H-LPS may play a critical role in inducing inflammatory reaction in human gastritis.
测定幽门螺杆菌(H. pylori)脂多糖(H-LPS)的生物学活性,了解H-LPS与人类慢性胃炎及消化性溃疡之间的病理相关性。
采用酚水法提取临床幽门螺杆菌菌株的H-LPS和大肠杆菌O55:B5菌株的脂多糖(E-LPS)。通过鲎试剂法、热原试验、家兔血压试验、家兔外周血单核细胞(PBMC)诱导试验、NIH 3T3成纤维细胞毒性试验及NIH小鼠致死试验检测H-LPS和E-LPS的生物学活性。以自制兔抗H-LPS血清为一抗,市售辣根过氧化物酶(HRP)标记羊抗兔血清为二抗,采用免疫组织化学法检测126例慢性胃炎患者(68例)或胃溃疡患者(58例)活检标本中的H-LPS。
H-LPS对成纤维细胞的毒性及对小鼠的致死性均弱于E-LPS。但两种脂多糖的鲎试剂凝聚能力相似(+/0.5 ng/mL)。注射H-LPS后0.5小时,3只家兔血压迅速下降。注射H-LPS后1.0小时,3只家兔中有2只血压降至零,导致这2只动物死亡。同组另1只家兔血压逐渐升高。注射E-LPS后0.5小时,3只家兔血压也迅速下降,然后维持在低水平约1.0小时。注射H-LPS或E-LPS后0.5小时,家兔PBMC数量显著增加。126份活检标本中H-LPS的总阳性率为60.3%(76/126)。慢性胃炎患者活检标本中H-LPS阳性率(50/68,73.5%)显著高于胃溃疡患者(26/58,44.8%)(χ²=10.77,P<0.01)。慢性浅表性胃炎患者活检标本中H-LPS阳性率(38/48,79.2%)和慢性活动性胃炎患者(9/10,90.0%)显著高于萎缩性胃炎患者(3/10,30.0%)(χ²=7.50 - 9.66,P<0.01)。
H-LPS的生物学活性弱于E-LPS,其降低家兔血压及诱导家兔PBMC的活性相对较强。H-LPS可能在人类胃炎炎症反应诱导中起关键作用。