Abe K, Okina A, Yano T, Gao C, Ohmori H, Ishibashi K, Nishiura T, Letić-Gavrilović A
Department of Oral Biochemistry, Fukuoka Dental College, Japan.
J Dent Res. 1998 Nov;77(11):1913-9. doi: 10.1177/00220345980770110801.
To identify salivary biomarkers of periodontal diseases, we used plaque-resistant and -susceptible rats as animal models. The levels of salivary cystatin S in saliva, salivary glands, and gingiva were tested in Nembutal-anesthetized young and adult plaque-resistant and -susceptible rats of both sexes with and without chronic treatment with isoproterenol. Isoproterenol was injected i.p. once a day for 4 or 6 consecutive days. Isoelectric focusing electrophoresis by the PhastSystem and the western blotting method were used to separate different proteins and to identify a salivary cystatin S band in these samples. The expression of salivary cystatin S mRNA was also determined by the northern blotting method. Depending upon the types of agonists, a few differences were observed in secretory functions between both strains of rats in both sexes, but the levels of salivary cystatin S in saliva elicited from the submandibular gland and in the extracts of the submandibular glands and gingiva were significantly higher in plaque-resistant rats when compared with those of plaque-susceptible rats in both sexes. However, no significant difference was seen between the strains after chronic treatment with isoproterenol. The N-terminal 26-amino-acid sequence of salivary cystatin S purified from submandibular saliva of plaque-resistant rats was identical with that purified from submandibular saliva of Sprague-Dawley rats subjected to chronic treatment with isoproterenol. The expression of salivary cystatin S mRNA was dramatic in the submandibular glands of the plaque-resistant rats and in the submandibular glands of Wistar rats subjected to chronic treatment with isoproterenol, but not in those of plaque-susceptible rats. These results suggest that salivary cystatin S might be a good biomarker in distinguishing between the two strains of rats and that its concentration is correlated with plaque resistance.
为了鉴定牙周疾病的唾液生物标志物，我们使用抗菌斑和易患菌斑的大鼠作为动物模型。在戊巴比妥麻醉下，对未接受和接受异丙肾上腺素慢性治疗的成年和幼年抗菌斑和易患菌斑的雌雄大鼠，检测其唾液、唾液腺和牙龈中唾液胱抑素S的水平。异丙肾上腺素腹腔注射，每天1次，连续注射4或6天。采用PhastSystem等电聚焦电泳和蛋白质印迹法分离不同蛋白质，并鉴定这些样品中的唾液胱抑素S条带。唾液胱抑素S mRNA的表达也通过Northern印迹法测定。根据激动剂类型，在雌雄两性的两种品系大鼠之间观察到一些分泌功能差异，但与易患菌斑的雌雄大鼠相比，抗菌斑大鼠下颌下腺分泌的唾液、下颌下腺提取物和牙龈中唾液胱抑素S的水平显著更高。然而，异丙肾上腺素慢性治疗后，品系间未见显著差异。从抗菌斑大鼠下颌下唾液中纯化的唾液胱抑素S的N端26个氨基酸序列，与经异丙肾上腺素慢性治疗的Sprague-Dawley大鼠下颌下唾液中纯化的序列相同。唾液胱抑素S mRNA在抗菌斑大鼠的下颌下腺以及经异丙肾上腺素慢性治疗的Wistar大鼠的下颌下腺中表达显著，但在易患菌斑的大鼠中不表达。这些结果表明，唾液胱抑素S可能是区分两种品系大鼠的良好生物标志物，其浓度与抗菌斑能力相关。
