Kudo K, Saito A, Sudo K, Adachi M, Ikai A, Ofuji Y, Tadeuchi Y, Yoshie H, Hara K
Nihon Shishubyo Gakkai Kaishi. 1988 Dec;30(4):1061-9. doi: 10.2329/perio.30.1061.
We examined in vitro the inhibitory effects of ovomacroglobulin on collagenolytic activity in Bacteroides gingivalis (B. gingivalis) culture supernatant, in human peripheral blood polymorphonuclear leucocytes (PMN), and in gingival crevicular fluid (GCF) from periodontitis patients. Measurement of collagenolytic activity was conducted with a CollagenoKit CLN-100 using FITC-conjugated type I collagen. The FITC-conjugated collagen was reacted with the sample in solution, and the residue was selectively degenerated at 35 degrees C and removed with ethanol. The fluorescence of the removed residue was then measured. The collagenolytic activity from B. gingivalis displayed dose dependent inhibition as high as 81.4% following addition of ovomacroglobulin at 224 micrograms/ml. The collagenolytic activity from human peripheral blood PMN showed, as a result of addition of 1,600 micrograms/ml of ovomacroglobulin, inhibition as high as 62.4%. The collagenolytic activity from human GCF, which was obtained from patients with different degrees of periodontal disease, exhibited as high as 71.0% inhibition after addition of 1,600 micrograms/ml ovomacroglobulin. Ovomacroglobulin showed almost the same level of inhibition obtained from alpha 2-macroglobulin, which was measured as a positive control. It was also recognized by SDS-PAGE that collagenolytic activity was inhibited after preincubation with added ovomacroglobulin. This collagenolytic activity, which dissolved the collagen substrate, was derived from B. gingivalis and human GCF. The above results demonstrate that ovomacroglobulin inhibits collagenolytic activity from B. gingivalis, human PMN, and human GCF.
我们在体外研究了卵巨球蛋白对牙龈卟啉单胞菌(牙龈卟啉单胞菌)培养上清液、人外周血多形核白细胞(PMN)以及牙周炎患者龈沟液(GCF)中胶原酶活性的抑制作用。使用FITC偶联的I型胶原,通过CollagenoKit CLN - 100测定胶原酶活性。将FITC偶联的胶原与溶液中的样品反应,残余物在35℃下选择性降解,然后用乙醇去除。随后测量去除残余物的荧光。在加入224微克/毫升的卵巨球蛋白后,牙龈卟啉单胞菌的胶原酶活性表现出高达81.4%的剂量依赖性抑制。加入1600微克/毫升的卵巨球蛋白后,人外周血PMN的胶原酶活性显示出高达62.4%的抑制。从不同程度牙周病患者获得的人GCF的胶原酶活性,在加入1600微克/毫升卵巨球蛋白后表现出高达71.0%的抑制。卵巨球蛋白显示出与作为阳性对照测量的α2 - 巨球蛋白几乎相同水平的抑制。通过SDS - PAGE还认识到,在与添加的卵巨球蛋白预孵育后,胶原酶活性受到抑制。这种溶解胶原底物的胶原酶活性源自牙龈卟啉单胞菌和人GCF。上述结果表明,卵巨球蛋白抑制牙龈卟啉单胞菌、人PMN和人GCF的胶原酶活性。