唾液蛋白质组学的研究进展

唾液作为一种成分复杂，在口腔内具有多种生物学功能的液体。它起到润滑、辅助消化、抗茵等诸多作用，而对唾液的功能和其分泌的分子机制的研究离不开对唾液蛋白质的研究。蛋白质组学作为大规模蛋白质分离和识别技术研究蛋白质组的一门学科，已经开始在很多领域得到应用，同时正逐渐成为唾液蛋白组研究中的新技术。本文就蛋白质组学在唾液中的蛋白全表达、生物标记应用等方面内容作一综述。     

唾液蛋白质组学的研究进展

唾液作为一种成分复杂,在口腔内具有多种生物学功能的液体.它起到润滑、辅助消化、抗菌等诸多作用,而对唾液的功能和其分泌的分子机制的研究离不开对唾液蛋白质的研究.蛋白质组学作为大规模蛋白质分离和识别技术研究蛋白质组的一门学科,已经开始在很多领域得到应用,同时正逐渐成为唾液蛋白组研究中的新技术.本文就蛋白质组学在唾液中的蛋白全表达、生物标记应用等方面内容作一综述.     

唾液蛋白质组学研究进展

唾液蛋白质组学的研究,对于寻找疾病分子标志物和药物靶标具有重要意义,对于疾病的诊疗具有难以估量的应用前景.本文就唾液蛋白质组学研究意义、研究方法、相关技术、应用研究、存在问题和发展趋势进行综述.     

腮腺非肿瘤性疾病混合唾液和腮腺液的细菌学研究

目的 探讨涎腺非肿瘤性疾病所引起的口腔微生态的变化。方法 ２０例患者,包括１１例舒格伦综合征（ＳＳ）、１０例腮腺良性肥大及８例慢性腮腺炎和１０名正常对照者。收集其混合唾液和腮腺液并进行口腔内５种常见致病菌的鉴定及细菌总量的测定。结果 ＳＳ、腮腺慢性炎症患者混合唾液中需氧菌、厌氧菌、兼性链球菌及放线菌总数较对照组明显升高,ＳＳ患者混合唾液中的变链菌、乳酸菌总数较对照组明显升高;２例患者腮腺导管内有不     

唾液蛋白质组学与口腔癌生物标志物

该文旨在研究人类唾液中的蛋白标志物，评价其对口腔鳞癌的潜在诊断价值。收集64例OSCC患者的唾液样本，并与健康者进行对照。应用鸟枪法（shotgun）蛋白质组学对16例实验组及配对对照组的蛋白质谱进行分析。对其余48例标本进行免疫测定，以进一步确认候选的生物标志物。采用ROC分析法评估发现的候选标志物对OSCC的诊断价值。     

基于唾液宏蛋白质组学的重度低龄儿童龋患者唾液微生物群落分析

［摘要］ 目的 采用宏蛋白质组学技术研究重度低龄儿童龋患者唾液微生物群落的特征。方法 采集重度低龄儿童龋及无龋儿童非刺激性唾液,提取唾液中的蛋白质、酶解形成多肽后进行质谱分析,对比微生物库分析唾液微生物群落的特征。结果 无龋儿童唾液微生物来源于19个门1 216个种,高丰度的微生物以变形菌门、厚壁菌门、拟杆菌门、放线菌门、梭杆菌门以及乳糖奈瑟氏菌、肺炎链球菌、干燥奈瑟氏菌、副流感嗜血杆菌、脑膜炎奈瑟氏菌、流感嗜血杆菌、浅黄奈瑟氏菌、淋病奈瑟氏菌、金氏金菌、黏膜奈瑟氏菌、多糖奈瑟氏菌等11种细菌为主;重度低龄儿童龋儿童唾液微生物来源于24个门1 698个种,高丰度的微生物以变形菌门、厚壁菌门、拟杆菌门、放线菌门、蓝藻菌门以及肺炎链球菌、乳糖奈瑟氏菌、干燥奈瑟氏菌、流感嗜血杆菌等4种细菌为主。结论 宏蛋白质组技术可以全面、快速分析口腔唾液微生物群落的构成。重度低龄儿童龋儿童唾液微生物群落结构相比无龋儿童更加复杂,这种复杂性可能与龋病的产生及唾液微生态失衡有关。     

变异链球菌的比较蛋白质组学研究进展

最近公布的变异链球菌全菌基因组序列,使得对变异链球菌全面的蛋白组图谱分析成为可能。变异链球菌的蛋白质组学研究将进一步揭示其致龋的分子机制。比较蛋白质组学是蛋白质组学的一个重要分支,因此下面就变异链球菌对酸反应的比较蛋白质组学研究和变异链球菌浮游态与生物膜态的比较蛋白质组学等研究,介绍比较蛋白质组学在变异链球菌中的研究进展。     

差异蛋白质组学常用技术及其在牙周炎研究中的应用

蛋白质组学是连接基因、蛋白质和生命活动研究的桥梁,差异蛋白质组学则是它的主要研究内容之一.利用其常用技术来检测牙周炎不同环境下的差异蛋白,将有助于对疾病的诊断和治疗.本文就差异蛋白质组学常用技术及其在牙周炎中的应用作一综述.     

大鼠腮腺及颌下腺唾液的收集

目的:探讨大鼠腮腺及颌下腺唾液的收集方法。方法:采用微型Lashley吸盘法收集大鼠腮腺唾液,经口内颌下腺导管口直接插管法收集颌下腺唾液。毛果芸香碱刺激唾液分泌,假设唾液的比重为1.0 g/cm3,以唾液重量代表其体积,记录唾液流率。结果:大鼠腮腺唾液流率(9.9±1.4)μL/m in,颌下腺导管插管唾液流率(19.9±10.8)μL/m in。结论:可以采用微型吸盘法收集大鼠腮腺唾液,经口内颌下腺导管口直接插管法收集大鼠颌下腺唾液。     

唾液蛋白质组学的研究进展与应用

唾液是人体及其他哺乳动物一种重要且必需的体液,近年,成为除血浆、血清、尿液以外另一种检测人体生理、病理状态的有效工具[1-2].唾液主要由腮腺、舌下腺、下颌下腺三对大唾液腺和大量口腔黏膜下的小唾液腺分泌,并含有部分非唾液腺来源成分,主要包括龈沟液、口腔固有菌群及其产物、上呼吸道分泌物、口腔屏障渗透而来的血浆及血细胞、脱落的口腔上皮细胞以及食物残渣等[3].安静状态下唾液成分主要来源于舌下腺和颌下腺,而刺激状态下则主要由腮腺分泌的唾液组成[4-5],这种差异在实验设计时可作为唾液收集形式的依据.     

口腔微生物宏基因组学总DNA制备的方法学初探

目的：比较不同时间段人类口腔唾液微生物总DNA制备方法。方法：采用酚氯仿抽提法、QIAamp DNA Micro Kit法制备健康人群不同时间段口腔唾液微生物总DNA。使用紫外分光光度计,PCR等鉴定提取的总DNA。结果：两种方法均成功制备了口腔微生物总DNA。不同方法提取的总DNA在片段大小,纯度,得率等方面存在差异。短期内不同时间段口腔唾液微生物总DNA无显著差异。结论：两种总DNA提取方法均可用于宏基因组学研究,可根据不同的实验目的,选择更适合的方法。同一供体短期内不同时间段的唾液可混合用于宏基因组学研究。     

Preliminary proton nuclear magnetic resonance studies of human saliva

Summary-Proton NMR spectra of gustatory stimulated healthy human whole, parotid and submandibular and sublingual saliva were measured. The typical patterns of their spectra were obtained. Among these three kinds of fluids the parotid saliva which contains preferentially serous saliva presented a relatively well resolved spectrum with satisfactory signal to noise ratio in a given short time(30min.). For eight subjects of parotid saliva collected in the afternoon the marked increase in signal intensity in the methyl and methylene proton region of proteins(peptides)was observed as compared with those collected in the morning, reflecting the circadian rhythms in the protein concentration in saliva. On the other hand the methyl peak of lactic acid presented the opposite tendency, which might be also correlated with the circadian rhythms of the glucose metabolism of the parotid gland. The proton NMR spectra of three patients suspected of salivary gland disease were different from those of healthy subjects.     

Variability of flow rate when collecting stimulated human parotid saliva

The aim of this study was to estimate the accuracy and reproducibility of citric‐acid‐stimulated parotid saliva sampling. In healthy volunteers a strong correlation (r² = 0.79) between flow rates from the left and right parotid gland was observed. In patients with Sjögren's syndrome this correlation (r² = 0.90) was even stronger. The intraindividual variation in healthy volunteers was 23.3 ± 5.9%. Increasing the number of collections did not reduce this variation significantly. In head and neck cancer patients, to estimate whether repeated measurements result in more reliable baseline values for use in clinical studies, repeated collections did not result in a significant reduction of intrapatient variation, similar to the results with the healthy volunteers. Thus, notwithstanding the good agreement between left and right flow rates, a high variation in parotid flow rates has to be considered when planning clinical trials evaluating the effects of treatment on salivary gland functioning.     

Secretion of glucose in human parotid saliva after carbohydrate intake

Abstract – The aims of the present investigation were, first, to follow the secretion of free glucose in parotid saliva in various subjects after a single oral intake of different carbohydrates, and second, to compare the salivary glucose concentration with the concentration in blood. Twenty healthy subjects, three women and 17 men, 20–35 yr of age, participated. They were asked not to eat or drink anything from 10 p.m. the night before the examination. 75 g of carbohydrate (glucose, fructose, or sucrose) dissolved in 300 ml water was ingested the next morning at 8 a.m. One experimental series with glucose was performed in triplicate in 10 of the subjects. Approximately 1.5 ml of citric acid-stimulated parotid saliva was collected before (0 min) and 15, 30, 45, 60, and 120 min after the intake. Salivary concentration of glucose was analyzed enzymatically. Most of the 0-min samples showed a variation in glucose concentration from 5 to 25 μmol/l. After the glucose, fructose, and sucrose intakes, the salivary glucose level increased about 2–4 times, especially in the 30-min samples. A large inter- as well as intra-individual variation was found both in the 0-min samples and in the samples collected after the different intakes. The correlation between the glucose concentration in saliva and blood was higher after than before the carbohydrate intakes.     

A modified device for collection and flow-rate measurement of submandibular-sublingual saliva

The aims of the present study were to measure stimulated submandibular-sublingual (SM-SL) salivary flow rate with a modified Block-Brottman collection device, and, further, to evaluate the reliability of measurements of stimulated SM-SL salivary flow rate by means of this modified Block-Brottman device, as compared to measurements of parotid flow rate using modified Carlson-Crittenden cups. Twenty-nine healthy female volunteers, aged 36±7 yr, were included. Saliva stimulation was achieved by application of a 3% citric acid solution to the rims of the tongue four times/min, for 3 s every 15 s. On 3 consecutive days, stimulated parotid and SM-SL salivas were collected for 2 min at 07.30, before breakfast (morning value), and at 10.00, 2 h after a standard breakfast (lunchtime values). SM-SL saliva was also collected on one occasion for 2 min ± 3. For parotid and SM-SL saliva, the mean stimulated flow rates were in the morning, 1.50±0.83 and 2.25±1.12 ml/min, and at lunchtime, 1.71±1.16 and 2.54±1.01 ml/min, respectively. For both salivas, lunchtime values were significantly higher than morning values by about 13–14%. Comparing parotid and SM-SL saliva flow rates, we found the SM-SL saliva flow rate to exceed the parotid flow rate by about 50% both in the morning and at lunchtime. Variations in flow rate were analyzed by means of ANOVA. Interindividual variance and variance between measurement days and times of day made up 88% of parotid and 83% of SM-SL total variance. By calculating the variation coefficient, we found this to be smaller for SM-SL salivary flow rate measurements as compared to parotid flow rate measurements. In conclusion, the results of the present study indicate that our method of collecting stimulated submandibular-sublingual saliva by means of a modified Block-Brottman collection device is as reliable as the method of collecting stimulated parotid saliva by means of modified Carlson-Crittenden cups, and is thus a useful method for future studies of changes in submandibular-sublingual salivary production.     

Inhibitory effects of whole and parotid saliva on immunomodulators

Based on the presence of cytokines in whole saliva and their association with resistance and susceptibility to infectious disease, the present study was designed to evaluate the diagnostic potential of a large panel of cytokines and chemokines in saliva. Despite the endogenous presence of Th1/Th2 and pro-inflammatory cytokines and several chemokines in whole and parotid saliva of most individuals tested, the detection of known concentrations of several recombinant cytokines and chemokines was inhibited immediately following their addition to each type of saliva. In contrast, purified immunoglobulins were unaffected by either whole or parotid saliva. Further studies revealed that the inhibition of immunoreactivity involved sequestration of the majority of cytokines affected and degradation of chemokines. These results suggest that absolute concentrations of cytokines/chemokines may not be fully detectable in saliva. Therefore, the diagnostic value of any cytokine/chemokine is questionable and should be evaluated independently as such.     

Film-forming properties and viscosities of saliva substitutes and human whole saliva

Hypo-salivation, related to medical remedies, is an increasing clinical problem. Studies report a weak correlation between subjective mouth dryness and objective sialometry. This indicates that both quantity and quality of saliva are important for the surface-associated functions of saliva, such as lubrication and hydration, to be expressed. Film-forming properties and viscosities of three saliva substitutes were compared to human saliva. Adsorption to surfaces was measured by ellipsometry, infrared spectroscopy and drop-volume technique. Viscosity measurements were carried out using an oscillating rheometer. Saliva, with the lowest viscosity value and the highest protein content, presented superior film retention on both hydrophilic and hydrophobic surfaces. The carboxymethylcellulose-based MAS 84 showed intermediate values of viscosity, poorest ability to reduce surface tension, and negligible film-forming capacity. The porcine mucin-based Saliva Orthana showed about twice the viscosity of saliva and film-forming capability on preferably hydrophobic substrates. Salinum, a linseed extract, possessed the highest viscosity value and an initial surface tension close to that of saliva. The film retention on hydrophilic surfaces was not as effective as for saliva. The results indicate that the film-forming capacity of saliva substitutes is a property also to be considered in the exploration of clinically effective artificial salivas.     

Parotid function and composition of parotid saliva among elderly edentulous African – American diabetics

This study examined parotid salivary flow rate and composition in three groups of differently treated diabetics and a control group of non-diabetics. The study population was composed of edentulous African-Americans at least 65 years of age. Group A was the control, Group B insulin-dependent (Regular Iletin, U-100 qd.). Group C controlled by oral medication (Tolbutamide, 500 mg qd.), and Group D was diet controlled. All diabetic patients had serum glucose values under 250 mg/dl. Salivary flow rates, secretory IgA. electrolytes (Na⁺, C1^-, K⁺, Ca⁺⁺) and total protein concentrations were evaluated. The results showed no significant differences between groups with respect to salivary flow rates, electrolytes and IgA concentrations. Additionally, all diabetic groups demonstrated a significantly lower salivary total protein concentration when compared to the controls. There appears to be no evident decrease in salivary flow rate in these three differently controlled diabetic groups compared with healthy non-diabetics.