Enhancement of Ca^<2+>-influx and prostaglandin E_2accumulation in cultured pig vascular smooth muscle cells infected with Salmonella Choleraesuis

猪霍乱沙门氏菌感染培养猪血管平滑肌细胞Ca^2内流和前列腺素E_2积累的增强

• 批准号: 08670316
• 负责人: DANBARA Hirohumi
• 金额: $ 1.47万
• 依托单位: Department of Microbiology School of Pharmaceutical Sciences Kitasato university
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 1996
• 资助国家: 日本
• 起止时间: 1996 至 1997
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-08670316/
• 关键词: Salmonella; Salmonella septicemia; Salmonells Choleraresuis; Smooth muscle cells; Arachidonic acid; Phospholipase A_2; Ca^<2+> influx; invA mutant; サルモネラ症; ホスフォリ-パーゼA_2; カルシウム阻害剤; チロシンキナーゼ阻害剤

Three clinical distinguishable forms of salmonellosis, enteric fever (typhoid fever and paratyphoid fever), septicemia and acute gastroenteritis occur in humans. Salmonella septicemia in humans are most commonly caused by S.Choleraesuis, a swine-adaped serotype of Salmonella. Our previous reports has shown that S.Choleraesuis strain RF-1 caused septicemia and killed pigs with severe hemorrhages on whole body. This results suggested that S.Choleraesuis caused lesions of the vascular tissues, which developed the hemorrhages in the infected pigs. In the present study, in vitro effects of S.Choleraesuis on cultured vascular smooth musle cells prepared from aortas of pigs were examined.S.Choleraesuis strain-RF-1 was observed to invade the vascular smooth muscle cells after 3 to 5 hours incubation in a time-dependent manner. RF-1 strain contracted the vascular smooth muscle cells in parallel with the increase of inyasion. This morphological change was diminished by incubation of the vascular … More smooth muscle cells with a invA mutant (a invasion-deficient derivative) of RE-1 strain. Accompanying the contraction, both influx of Ca^<2+> and activity of phopholipase A_2(PLA_2) activity were increased in the cells incubated with RF-1 atrain, but not with invA mutant, suggesting that the arachidonic acid cascades were activated by the invasion of S.Choleraesuis. Then we analyzed the metabolites of arachidonic acid, prostaglandins (PGs) and leukotriens (LTs), in the vascular smooth muscle cells. It was demonstrated that the cellular contents of PGE_2 remarkably increased in the cells by the invasion of RF-1 strain. However, the contents of neither LTC_4, LTD_4 nor LTE_4 were influenced. Our results showed that invasion of S.Choleraesuis enhanced Ca^<2+> influx and activated PLA_2 resulting in the accumulation of cellular PGE_2 in the vascular smooth muscle cells. The increase of PGE_2 could cause the contraction of vascular smooth muscle cells, which may lead to the lesions of the vascular tissues of pigs infected with S.Choleraesuis. Less

沙门氏菌病的三种临床可区分的形式，即肠热病（伤寒和副伤寒）、败血症和急性胃肠炎发生在人类中。人类沙门氏菌败血症最常见的是由猪霍乱沙门氏菌引起的，这是一种猪适应的沙门氏菌血清型。我们以前的报道表明，猪霍乱沙门氏菌RF-1株引起猪败血症，并造成猪全身严重损伤。结果提示，猪霍乱弧菌可引起血管组织病变，进而引起血管病变。本研究观察了猪霍乱沙门氏菌对体外培养的猪血管平滑肌细胞的作用，结果表明，猪霍乱沙门氏菌RF-1株在体外培养3 ~ 5 h后即可侵入血管平滑肌细胞，并呈时间依赖性。RF-1株使血管平滑肌细胞收缩，同时使平滑肌细胞收缩率增加。这种形态学变化被血管的孵育所减弱。 ...更多信息 具有RE-1菌株的invA突变体（侵袭缺陷衍生物）的平滑肌细胞。在RF-1 atrain孵育的细胞中，Ca^<2+>内流和磷脂酶A_2（PLA_2）活性均增加，而invA突变体则无此变化，提示花生四烯酸级联反应被猪霍乱链球菌入侵激活。然后分析了花生四烯酸在血管平滑肌细胞中的代谢产物--花生四烯酸苷（PGs）和白三烯（LTs）。结果表明，RF-1株侵入后，细胞内PGE_2含量明显增加。但对LTC_4、LTD_4和LTE_4含量无影响。结果表明，猪霍乱弧菌入侵后，血管平滑肌细胞内Ca^<2+>内流增加，PLA_2活化，PGE_2大量积聚。PGE_2升高可引起血管平滑肌细胞收缩，从而导致猪霍乱弧菌感染后血管组织病变。少

项目成果

小林温,長井正昭,檀原宏文: "Salmonella Choleraesuis感染における平滑筋細胞内ホスオ-リパーゼA_2活性とサイトカラミンとの関係" 日本細菌学雑誌. 52・1. 324- (1997)

Atsushi Kobayashi、Masaaki Nagai、Hirofumi Danhara：“猪霍乱沙门氏菌感染中平滑肌细胞内磷脂酶 A_2 活性与细胞胺的关系”日本细菌学杂志 52・1。