AQP3、7、9在人下颌下腺的免疫组化定位

采用免疫组化ABC法检测源自10例颈淋巴清扫患者的正常下颌下腺组织AQP3、7、9蛋白的表达。AQP3在浆液性腺泡和黏液性腺泡细胞均有表达,分布于腺泡细胞的基底膜和侧膜,在浆液性腺泡细胞中的表达明显高于黏液性腺泡细胞,导管系统未见AQP3表达;AQP7在极少数肌上皮细胞有表达;AQP9在部分浆液性腺泡腔、混合性腺泡腔以及部分闰管管腔和浆液性半月体上有表达。     

水通道蛋白-1,5在干燥综合征小鼠下颌下腺中的表达

目的：探讨水通道蛋白-1和-5（AQP1、AQP5）在干燥综合征（Sjoegren syndrome，SS）小鼠下颌下腺中的表达及其意义。方法：选用BALB／c小鼠10只，用完全弗氏佐剂＋同种鼠下颌下腺抗原诱导、使之产生在临床表现、病理损害和免疫特征上都类似于人SS的动物模型．对照组10只均注射等体积的生理盐水。用免疫组化和Western印迹方法研究SS小鼠下颌下腺AQP1、AQP5表达的变化特点。结果：SS小鼠下颌下腺组织中AQP1和AQP5的表达量均显著减少（P〈0．01），主要分布在导管部位：AQP5在腺泡顶膜表达减少，而基侧膜的表达增强。结论：AQP1和AQP5表达量减少以及分布部位的异常改变，可能是SS患者口干的重要原因之一。     

水通道蛋白在干燥综合征患者唇腺中表达的实验研究

目的 :初步探讨水通道蛋白 -1和 -5 (AQP1、AQP5)在干燥综合征 (Sj grensyndrome ,SS)患者唇腺中的水代谢障碍发生机制 ,为SS涎腺的基因治疗提供理论依据。方法 :用免疫组化和Westernblot方法研究SS患者唇腺AQP1、AQP5表达变化特点。结果 :SS患者唇腺组织中AQP1和AQP5的表达量均显著减少 (P <0 .0 1) ,主要分布在导管部位 ,AQP5在腺泡顶膜表达减少 ,而基侧膜的表达增强。结论 :AQP1和AQP5表达量减少以及分布部位的异常改变是SS患者口干的重要原因之一。     

腺病毒介导CTLA4Ig对舍格伦综合征小鼠颌下腺AQP5表达的影响

目的：观察CTLA4Ig-重组腺病毒载体（Ad-CTLA4Ig）对SS颌下腺AQP5表达的影响。方法：采用完全弗氏佐剂＋同种鼠颌下腺抗原激发诱导小鼠舍格伦综合征（SS）。实验组在激发前30min予以Ad-CTLA4Ig腹腔注射,未转CTLA4IgcDNA的腺病毒载体（Adv）及SS作对照。比较各组动物颌下腺形态学改变,并采用Western blot分析SS小鼠颌下腺AQP5表达变化。结果：CTLA4Ig-重组腺病毒实验组颌下腺病理学改变不明显。并且删表达量明显高于对照组（P〈0．05）。结论：Ad-CTLA4Ig可正向调节小鼠颌下腺AQP5的表达,并可有效抑制小鼠合格伦综合征的发生。     

黄芪丹参对干燥综合征模型大鼠颌下腺组织的水通道蛋白-5表达的影响

目的 探讨黄芪合并丹参对干燥综合征模型大鼠颌下腺组织水通道蛋白-5(AQP5)的影响。方法 自身同种鼠抗原合并百白破疫苗加强免疫法免疫模型动物;用Western blot分析SS大鼠颌下腺AQP5表达的影响。结果 免疫动物出现类似SS的病理改变,存在大量淋巴细胞浸润,提示免疫造模成功。Western blot检测显示各组均出现分子量为42 KD的条带。空白对照组AQP5表达值正常(1.35±0.13),造模生理盐水组AQP5表达值(1.03±0.08)明显减少,造模中药高剂量组AQP5表达值较造模生理盐水组明显增强(1.31±0.15)。与空白对照组比较,造模生理盐水组AQP5表达显著减少(P<0.05);与造模生理盐水组比较造模中药高剂量组AQP5有表达显著增加(P<0.01)。结论 唾液流量的增加、颌下腺指数降低提示中药黄芪、丹参有改善唾液腺分泌唾液的功能;Western blot检测显示黄芪、丹参通过增强颌下腺水分子通道的释放,上调AQP5的表达,扩大颌下腺水分子的滤过,缓解口腔干燥的症状。?     

α1-肾上腺素受体在颌下腺的表达及苯肾上腺素对唾液分泌调节的动物实验

目的研究α-肾上腺素受体亚型在家兔颌下腺的表达和分布，及其激动剂——苯肾上腺素促家兔颌下腺唾液分泌的相关机制。方法应用RT—PCR和Western blot检测家兔正常颌下腺α1-肾上腺素受体亚型mRNA及蛋白质的表达；免疫组化法检测颌下腺α1-肾上腺素受体亚型的分布及水通道蛋白5的表达；经颌下腺导管插管给予（1×10^-8）-（1×10^-6）moL／L的苯肾上腺素，观察家兔心率和血压的变化及促颌下腺唾液分泌的量效关系。结果家兔颌下腺有α1A-α1B和α1D-肾上腺素受体3种亚型的mRNA及蛋白质的表达，并广泛分布于导管和腺泡细胞的胞膜及胞质中。给予1×10^-7moL／L苯肾上腺素7d，促进颌下腺唾液分泌增加，而对心率、血压无明显影响；水通道蛋白5在腺泡及导管细胞的顶膜和侧膜的表达增加。结论家兔颌下腺存在α1A-、α1B-和α1D-肾上腺素受体3种亚型的mRNA和蛋白质的表达。经颌下腺导管给予低剂量苯肾上腺素促进唾液分泌是安全有效的；水通道蛋白5的表达增加可能与苯肾上腺素促唾液分泌的机制有关，该研究为临床治疗领下腺功能低下提供了初步依据。     

HBD-1和HBD-2在口腔鳞癌患者颌下腺中的表达

目的：研究口腔鳞癌患者颌下腺细胞系（HSG）及颌下腺组织中HBD-1和HBD-2的表达和分布,为恶性肿瘤患者预防感染,提高机体免疫力提供理论依据。方法：应用逆转录-聚合酶链式反应（RT-PCR）法观察HSG中HBD-1和HBD-2的表达;利用免疫组织化学方法观察6例口腔鳞癌颌下腺手术标本中HBD-1和HBD-2在腺泡和导管中的分布。结果：HSG中可检测出HBD-1和HBD-2的表达;颌下腺腺泡和导管中都有HBD-1和HBD-2分布。结论：口腔鳞癌患者颌下腺具有一定的免疫防御能力,颌下腺导管与恶性肿瘤抵御感染有密切关系。     

SD大鼠下颌下腺主导管结扎损伤及再通后的组织学观察

目的:建立主导管结扎的SD大鼠下颌下腺组织损伤模型,观察主导管结扎及再通后腺体的组织学变化。方法:双重结扎SD大鼠下颌下腺主导管。部分腺体在6 d后摘取;其他腺体于6 d后松开结扎使再通,并于4、6、12、24 d后处死动物,取出腺体。通过HE及免疫组织化学染色观察其组织化学变化。结果:主导管结扎6 d后,腺体中腺泡消失,导管样结构增生,导管细胞表达CK19;在增生导管及周围见α6β1、LN阳性细胞。松开结扎再通的导管在6 d后有成熟的腺泡细胞出现,24 d后有大量成熟腺泡形成,Amylases、PAS(periodic acid-schiff)、AQP-5均在腺泡细胞中表达。结论:导管结扎损伤后腺体腺泡萎缩,导管上皮增生。松开结扎再通后腺泡细胞再生、功能恢复。     

c-Met在小鼠颌下腺胚胎发育中的表达

目的探讨c-Met在小鼠颌下腺发育不同阶段的表达特点。方法制作ICR小鼠颌下腺不同发育阶段的冰冻组织切片,利用免疫组织化学方法对小鼠颌下腺自蕾状期至出生后两天不同发育阶段c-Met的表达情况进行了研究。结果 c-Met在小鼠颌下腺发育的蕾状期上皮中表达呈明显阳性,在假腺管期早期表达减弱,而在假腺管期晚期的上皮突起中表达增强。在微管期的分枝状上皮条索和顶端胚芽中呈不对称表达,周围细胞表达较强而中心细胞表达较弱。在终末分化期的腺泡、闰管的部分上皮细胞表达,在导管上皮中持续性强表达。结论 c-Met在小鼠颌下腺发育的细胞增殖、分支形态发生及腺泡和导管的形成中可能扮演重要角色。     

颌下腺多形性腺瘤及其复发的临床研究

目的:评价颌下腺多形性腺瘤及其复发的临床特点及原因。方法:对8例颌下腺多形性腺瘤复发的临床特点和24例颌下腺手术野术中脱落细胞学进行分析及检测。结果:本院颌下腺多形性腺瘤术后有3例复发(3/366例,0.8%),另5例复发性颌下腺多形性腺瘤均为外院转入我院。8例病例复发时间均在1年以上,其中1例第2次复发时间为第1次复发术后5个月。1例复发性颌下腺多形性腺瘤术中见瘤体深叶处存留少许腺细胞,7例无腺细胞残留。术中脱落细胞显示8例原发性肿瘤手术野冲洗前有3例检测到腺泡细,5例为血液细胞,冲洗后2例为腺泡细胞,6例为血液细胞,8例复发性肿瘤冲洗前后均为1例腺泡细胞,7例血液细胞。8例非瘤性病变冲洗前1例为腺泡细胞,7例血液细胞,冲洗后均为血液细胞,3组之间无显著性差异(P>0.05)。结论:颌下腺多形性腺瘤复发罕见,可能与细胞学穿刺及瘤旁腺泡有关。     

Relationship between aquaporin‐5 expression and saliva flow in streptozotocin‐induced diabetic mice?

Oral Diseases (2012) 18 , 501–505 Objective: To investigate the expression and distribution of AQP5 in submandibular acinar cells from sham‐ and streptozotocin (STZ)‐treated mice in relation to the salivary flow. Methods: Mice were sham or STZ injected. Distribution of AQP5 subcellular expression in submandibular glands was determined by immunohistochemistry. AQP5 labelling indices (LI), reflecting AQP5 subcellular distribution, were determined in acinar cells. Western blotting was performed to determine the expression of AQP5 in submandibular glands. Blood glycaemia and osmolality and saliva flow rates were also determined. Results: AQP5 immunoreactivity was primarily located at the apical and apical‐basolateral membranes of submandibular gland acinar cells from sham‐ and STZ‐treated mice. No significant differences in AQP5 protein levels were observed between sham‐ and STZ‐treated mice. Compared to sham‐treated mice, STZ‐treated mice had significant increased glycaemia, while no significant differences in blood osmolality were observed. Saliva flow rate was significantly decreased in STZ‐treated mice as compared to sham‐treated mice. Conclusions: In STZ‐treated mice, significant reduction in salivary flow rate was observed without any concomitant modification in AQP5 expression and localization.     

Effects of phenylephrine on transplanted submandibular gland

Autotransplantation of the submandibular gland is a potential treatment for severe kerato-conjunctivitis sicca. However, one of the major barriers to this procedure is that secretions from the transplanted gland decrease shortly after the operation, which may lead to obstruction of Wharton's duct, or even to transplantation failure. Using a rabbit model, we investigated whether phenylephrine could improve the secretion from the transplanted gland. We found that phenylephrine treatment significantly reversed the decrease in salivary secretion after transplantation, enhanced the expressions of alpha1A-, alpha1B-, and alpha1D-adrenoceptor mRNA, and ameliorated atrophy of acinar cells. Furthermore, phenylephrine also induced translocation of aquaporin-5 from the cytoplasm to the apical membrane, and increased the levels of phospho-ERK1/2, ERK1/2, phospho-PKCzeta, and PKCzeta in the transplanted gland. These results indicate that phenylephrine treatment moderates structural injury and improves secretory function in the transplanted submandibular gland through promoting alpha1-adrenoceptor expression and post-receptor signal transduction.     

Cell surface markers CD44 and CD166 localized specific populations of salivary acinar cells

Oral Diseases (2012) 18 , 162–168 Objective: Experimental approaches tested to date for functional restoration of salivary glands (SGs) are tissue engineering, gene transfer, and cell therapy. To further develop these therapies, identifying specific cell surface markers for the isolation of salivary acinar cells is needed. To test a panel of cell surface markers [used in the isolation of mesenchymal stem cells, (MSCs)] for the localization of salivary acinar cells. Materials: Human submandibular and parotid glands were immunostained with a panel of MSC markers and co‐localized with salivary acinar cell differentiation markers [α‐amylase, Na‐K‐2Cl cotransporter‐1, aquaporin‐5 (AQP5)]. Additional cell markers were also used, such as α‐smooth muscle actin (to identify myoepithelial cells), cytokeratin‐5 (basal ductal cells), and c‐Kit (progenitor cells). Results: CD44 identified serous acini, while CD166 identified mucous acini. Cytokeratin‐5 identified basal duct cells and 50% of myoepithelial cells. None of the remaining cell surface markers (Stro‐1, CD90, CD106, CD105, CD146, CD19, CD45, and c‐Kit) were expressed in any human salivary cell. Conclusions: CD44 and CD166 localized human salivary serous and mucous acinar cells, respectively. These two cell surface markers will be useful in the isolation of specific populations of salivary acinar cells.     

Immunohistochemical localization of lectins in normal salivary glands and mucoepidermoid carcinoma

5 kinds of Lectins (ConA, PNA, RCA, SBA and WGA) were used to observe the immunochemical localization of lectins in the cells of normal salivary glands and mucoepidermoid carcinoma. The result shows that in normal salivary glands ConA staining was found only in duct cells and serous acinar cells. Intercalated duct cells and striated duct cells of parotid and submandibular gland displayed RCA and WGA staining. The duct cells of sublingual gland and palatine gland did not react with lectin RCA. The ductal and acinar cells of normal salivary gland were not stained by PNA. Serous acinar cells of sublingual gland could be stained by SBA but mucous cells were negative. In tumor cells, ConA, RCA, WGA showed extensive staining which suggests the tumor cells contain similar glycoproteins as the normal duct cells. PNA was found in squamous epithelial cells but not in glandular epithelium. Its presence in tumor cells may indicate the degree of differentiation of these cells. Epidermoid and mucous cells of the tumor were also stained by 54 Kd antikeratin antibody. The results suggest this tumor may originate from the ductal cells of salivary gland.     

Cell death and cell proliferation in the regeneration of atrophied rat submandibular glands after duct ligation

BACKGROUND: The present study aimed to clarify the proliferation and apoptosis of parenchymal cells during regeneration of rat submandibular glands following atrophy. METHODS: Atrophy of the right submandibular gland of rats was induced by excretory duct ligation at the hilum with metal clips, which were removed 1 week (day 0) after ligation. The right submandibular glands were collected from 0 to 14 days after removal of the clips and investigated using immunohistochemistry for proliferating cell nuclear antigen (PCNA) as a marker of proliferating cells, terminal deoxynucleotidyl transferase (TdT)-mediated dUTP-digoxigenin nick end labeling (TUNEL) as a marker of apoptotic cells, and transmission electron microscopy (TEM). RESULTS: After 1 week of ligation, there were many remaining ducts and a few acini in the atrophic glands. At day 3 after discontinuing the ligation, newly formed acini appeared and thereafter increased in number and maturity. Many residual and newly formed acinar cells showed positive reaction to PCNA especially at days 4 and 5. The PCNA-positive duct cells decreased in number with the regeneration. A few TUNEL-positive acinar and duct cells were identified during regeneration. Mitosis and apoptosis of parenchymal cells were also identified by TEM. CONCLUSIONS: During regeneration of the submandibular gland after atrophy, both residual and newly formed acinar cells proliferate actively. There is also apoptosis of parenchymal cells; however, the significance of apoptosis is low.     

Autonomic Nerve-Regulated AQP5 Distribution in Salivary Glands and AQP5 Release into Saliva

The salivary glands are innervated by autonomic nerves. Upon activation of M₃-muscarinic acetylcholine receptors or α₁-adrenergic receptors, cytosolic aquaporin-5 (AQP5) is translocated from intracellular endosomes to the apical plasma membrane (APM) together with lipid rafts in the parotid acinar and duct cells. Some AQP5 is dissociated from the lipid rafts to non-rafts in the APM and the dissociated AQP5 is released into the saliva from the APM. Other AQP5 associated with lipid rafts is internalized and recycled together with the lipid rafts. The subcellular distribution of AQP5 in the salivary glands and its release into the saliva are regulated by autonomic nerves. This review focuses on autonomic nerve-regulated AQP5 trafficking in the parotid glands and AQP5 release into the saliva.