Lundy F T, Chalk R, Lamey P J, Shaw C, Linden G J
School of Dentistry, Queen's University, Belfast, Northern Ireland, UK.
J Clin Periodontol. 2001 Dec;28(12):1172-7. doi: 10.1034/j.1600-051x.2001.281213.x.
The protein components of GCF can be separated by reverse-phase microbore HPLC on a C18 column with detection on the basis of 214 nm absorbance. A single major symmetrical protein peak eluting with a retention time of 26 min (50% acetonitrile) was evident in gingival crevicular fluid (GCF) from periodontitis patients but not in healthy GCF. This protein was identified as human MRP-8 by N-terminal amino acid sequencing and liquid chromatography quadropole mass spectrometry.
To quantify the amount of MRP-8 detectable in GCF from individual healthy, gingivitis and periodontitis affected sites and to study the relationship, if any, between the levels of this responsive protein and periodontal health and disease.
GCF was sampled (30 s) from healthy, gingivitis, and periodontitis sites in peridontitis subjects (n=15) and from controls (n=5) with clinically healthy gingiva and no periodontitis. Purified MRP-8 was sequenced by Edmann degradation and the phenylthiohydantoin (PTH) amino acid yield determined (by comparison of peak area with external PTH amino acid standards). This value was subsequently used to calculate the relative amount of protein in the peak eluting with a retention time of 26.0 min (MRP-8) in individual GCF chromatograms.
Higher levels of MRP-8 were detected in inflammatory sites: periodontitis 457.0 (281.0) ng; gingivitis 413.5 (394.5) ng compared with periodontally healthy sites in diseased subjects 14.6 (14.3) ng and in controls 18.6 (18.5) ng, p=0.003. There was at least 20-fold more MRP-8 in the inflammatory compared with the healthy sites studied.
The preliminary data indicate that MRP-8 is present in GCF, with significantly greater amounts present at diseased than healthy sites. A systematic study of the relationship of this protein to periodontal disease could prove useful in further clarifying whether MRP-8 could be a reliable GCF biomarker of gingivitis and periodontitis.
龈沟液（GCF）的蛋白质成分可通过反相微孔高效液相色谱法在C18柱上进行分离，并基于214nm吸光度进行检测。在牙周炎患者的龈沟液（GCF）中，有一个主要的对称蛋白峰在保留时间为26分钟（50%乙腈）时洗脱出来，但在健康龈沟液中未出现。通过N端氨基酸测序和液相色谱四极杆质谱法，该蛋白被鉴定为人髓样相关蛋白8（MRP-8）。
量化在个体健康、牙龈炎和牙周炎患部的龈沟液中可检测到的MRP-8的量，并研究这种反应性蛋白水平与牙周健康和疾病之间的关系（如果存在的话）。
从牙周炎患者（n = 15）的健康、牙龈炎和牙周炎部位以及临床牙龈健康且无牙周炎的对照组（n = 5）中采集龈沟液（30秒）。通过埃德曼降解法对纯化的MRP-8进行测序，并测定苯硫代乙内酰脲（PTH）氨基酸产量（通过将峰面积与外部PTH氨基酸标准进行比较）。该值随后用于计算个体龈沟液色谱图中保留时间为26.0分钟时洗脱峰（MRP-8）中蛋白质的相对量。
在炎症部位检测到较高水平的MRP-8：牙周炎为457.0（281.0）ng；牙龈炎为413.5（394.5）ng，而患病个体中牙周健康部位为14.6（14.3）ng，对照组为18.6（18.5）ng，p = 0.003。与所研究的健康部位相比，炎症部位的MRP-8至少多20倍。
初步数据表明MRP-8存在于龈沟液中，患病部位的含量明显高于健康部位。对该蛋白与牙周疾病关系的系统研究可能有助于进一步阐明MRP-8是否可能是牙龈炎和牙周炎的可靠龈沟液生物标志物。
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