Komine Yumiko, Kuroishi Toshinobu, Kobayashi Jin, Aso Hisashi, Obara Yoshiaki, Kumagai Katsuo, Sugawara Shunji, Komine Ken-ichi
Intelligent Cosmos Research Institute Corporation, Sendia, Japan.
J Vet Med Sci. 2006 Jul;68(7):715-23. doi: 10.1292/jvms.68.715.
Elastase activity and concanavalin A (Con A) low affinity bovine lactoferrin (bLf) molecule were detected in mammary gland secretions (MGSs) from mammary glands (MGs) with clinical staphylococcal mastitis. Changes in clinical symptoms correlated with increases in both elastase activity and the concentration of Con A low-affinity Lf in MGSs from mastitic MGs. Bovine Lf treated with elastase (elastase-Lf) showed various small bLf molecules and the same image on Con A two-dimensional immunoelectrophoresis as low Con A affinity bLf in MGSs. We confirmed the presence of four common bLf peptides for the elastase-bLf and low Con A affinity bLf molecules in mastitic MGSs, and synthesized four peptides. Strong mRNA expression of interleukin-6 (IL-6), tumor necrosis factor alpha (TNFalpha), interleukin-8 (IL-8), and monocyte chemotactic protein-1 (MCP-1) was induced in bovine mammary epithelial cells on stimulation with low Con A affinity bLf, elastase-bLf, and GQRDLLFKDSAL, a synthesis bLf peptide based on nuclear factor kappa B (NFkappaB) activation. These results suggest that bLf was cleaved by elastase, and that this cleavage changed the physical function of Lf. Our results indicate that elastase induced production of low Con A affinity bLf, including the bLf peptide GQRDLLFKDSAL, and had an inflammatory effect on staphylococcal mastitis.
在患有临床型葡萄球菌性乳腺炎的乳腺的乳腺分泌物（MGSs）中检测到了弹性蛋白酶活性和伴刀豆球蛋白A（Con A）低亲和力的牛乳铁蛋白（bLf）分子。临床症状的变化与乳腺炎乳腺的MGSs中弹性蛋白酶活性和Con A低亲和力Lf浓度的增加相关。用弹性蛋白酶处理的牛Lf（弹性蛋白酶-Lf）在Con A二维免疫电泳上显示出各种小的bLf分子，并且与MGSs中低Con A亲和力的bLf具有相同的图像。我们证实了在乳腺炎MGSs中弹性蛋白酶-bLf和低Con A亲和力bLf分子存在四种常见的bLf肽，并合成了四种肽。在用低Con A亲和力bLf、弹性蛋白酶-bLf和基于核因子κB（NFκB）激活的合成bLf肽GQRDLLFKDSAL刺激时，牛乳腺上皮细胞中白细胞介素-6（IL-6）、肿瘤坏死因子α（TNFα）、白细胞介素-8（IL-8）和单核细胞趋化蛋白-1（MCP-1）的mRNA表达强烈。这些结果表明bLf被弹性蛋白酶切割，并且这种切割改变了Lf的物理功能。我们的结果表明弹性蛋白酶诱导产生低Con A亲和力的bLf，包括bLf肽GQRDLLFKDSAL，并且对葡萄球菌性乳腺炎具有炎症作用。
