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首页> 外文期刊>Infection and immunity >Bacterial phospholipase C upregulates matrix metalloproteinase expression by cultured epithelial cells.
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Bacterial phospholipase C upregulates matrix metalloproteinase expression by cultured epithelial cells.

机译：细菌磷脂酶C通过培养的上皮细胞上调基质金属蛋白酶的表达。

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Phospholipase C (PLC) is a putative virulence factor of several pathogenic bacteria. We studied if exogenous PLC would perturb epithelial behavior in infected tissues. Gelatin and casein zymography of cell culture medium indicated that the broad-spectrum PLC of Bacillus cereus induced matrix metalloproteinase (MMP) production in epithelial cells of human skin (NHEK), human gingiva (HGE), and porcine periodontal ligament (PLE). In all three cell types, the strongest increase (ninefold) at 0.1 U/ml was seen in the MMP-9 (92-kDa gelatinase) activity, and the effect was dose dependent in the range of 0.1 to 1.0 U/ml. A relatively weaker increase (twofold) in MMP-2 (72-kDa gelatinase) was also observed in each cell type. PLC induction of MMP-3 (48-kDa stromelysin) was also seen in NHEK and HGE on gelatin and more sensitively for PLE by casein zymography (fivefold). Total gelatinolytic activity as measured by degradation of 14C-labeled denatured type I collagen increased by about 18-fold (NHEK), 12-fold (HGE), and 14-fold (PLE). Northern analysis showed a clear increase in the MMP-9, and a minor increase in MMP-3 mRNA levels but no significant increase in MMP-2 mRNA levels. Further studies with PLE revealed that MMP-9 induction by PLC progressively increased with the length of cell culture time in the absence of serum. PLC induction of MMPs was polar, with MMP-9 and MMP-3 secreted primarily in the apical direction and MMP-2 secreted mainly in the basal direction. The PLC effect was blocked by neomycin, an inhibitor of the phosphoinositol signal pathway. No significant effects were observed in MMP expression with the calcium ionophore A23187 or phospholipase A2. Morphologically, PLC treatment resulted in reduced contacts between the cultured cells and loss of the cell surface microvilli. These results suggest that PLC secreted by bacterial pathogens may disrupt epithelium of infected tissue and increase the subepithelial tissue destruction through induction of MMPs.
机译：磷脂酶C（PLC）是几种致病细菌的推定毒力因子。我们研究了外源性PLC是否会干扰感染组织的上皮行为。细胞培养基的明胶和酪蛋白酶谱分析表明蜡状芽孢杆菌的广谱PLC诱导人皮肤（NHEK），人牙龈（HGE）和猪牙周膜（PLE）上皮细胞中基质金属蛋白酶（MMP）的产生。在所有三种细胞类型中，MMP-9（92-kDa明胶酶）活性均以0.1 U / ml的幅度最大增强（九倍），且其作用在0.1至1.0 U / ml范围内。在每种细胞类型中，还观察到MMP-2（72-kDa明胶酶）相对较弱的增加（两倍）。在明胶中的NHEK和HGE中也观察到PLC诱导MMP-3（48 kDa溶血素），对酪蛋白酶谱（5倍）对PLE更敏感。通过降解14C标记的变性I型胶原蛋白测得的总明胶分解活性增加了约18倍（NHEK），12倍（HGE）和14倍（PLE）。 Northern分析显示MMP-9明显增加，MMP-3 mRNA水平略有增加，但MMP-2 mRNA水平无明显增加。 PLE的进一步研究表明，在无血清的情况下，PLC诱导的MMP-9随着细胞培养时间的延长而逐渐增加。 MMP的PLC诱导是极性的，MMP-9和MMP-3主要在根尖方向分泌，MMP-2主要在基部方向分泌。新霉素（一种磷酸肌醇信号途径的抑制剂）阻断了PLC的作用。用钙离子载体A23187或磷脂酶A2在MMP表达中未观察到显着影响。从形态上讲，PLC处理导致培养细胞之间的接触减少以及细胞表面微绒毛的丧失。这些结果表明细菌病原体分泌的PLC可能通过诱导MMPs破坏感染组织的上皮并增加上皮下组织的破坏。

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