丹地琼玉颗粒对大鼠离体颌下腺唾液分泌的影响

<正>目的:利用离体大鼠颌下腺灌流平台,观察丹地琼玉颗粒(Dan Di Qiong Yu granule,DDQY)对唾液分泌的影响。方法:健康雄性Wistar大鼠,体重200～220g,用苯巴比妥钠腹腔注射麻醉。     

养阴益气活血方对干燥综合征NOD小鼠颌下腺BCL-6和CXCR5表达的影响

目的:研究滤泡辅助T细胞(Tfh)相关分子BCL-6和CXCR5表达与干燥综合征(SS)的相关性,探讨养阴益气活血方治疗SS的可能机制。方法:将NOD小鼠随机分为4组,即模型组、中药组、西药组、结合组,分别灌服等量的去离子水、中药养阴益气活血方2.25 g/ml、羟氯喹3 mg/ml、中药加羟氯喹,灌胃、给药8周后解剖摘取组织计算颌下腺指数,HE染色观察颌下腺病理形态,免疫组化检测颌下腺Tfh特异分子BCL-6、CXCR5的表达。结果:小鼠颌下腺指数比较模型组与各用药组之间差异无显著统计意义(P>0.05);而结合组显著高于西药组(P<0.05); HE病理结果显示各组淋巴细胞浸润和腺泡结构破坏不等,颌下腺组织病理评分结合组显著低于模型组(P<0.05),其他组之间差异无显著统计学意义;小鼠颌下腺组织BCL-6和CXCR5的表达中药组和结合组显著低于模型组(分别为P<0.01,P<0.05),在BCL-6中药组和结合组还显著低于西药组(P<0.01)。结论:Tfh细胞及其相关分子BCL-6、CXCR5可能参与干燥综合征的发病,养阴益气活血方可能是通过...     

生长抑素对五肽胃泌素促进大鼠离体血管灌流全胃酸分泌功能的影响

本工作利用大鼠离体全胃血管灌流制备,在去除中枢神经系统及体液因素影响条件下,研究生长抑素(SOM)对五肽胃泌素(G-5)促进胃酸分泌功能的影响及其机制。以2ml/-min恒速从腹腔动脉灌流充以95％O_2、5％CO_2的Krebs-Ringer缓冲液,从肝门静脉收集血管灌流液,用放射免疫法分析血管流出液中前列腺素(PG)含量变化,胃腔以2ml/min恒速灌流充以100％O_2的蒸馏水,用0.01N NaOH溶液滴定分析胃酸排出量变化,结果如     

颌下腺注射间充质干细胞治疗干燥综合征的机制研究

目的探索干燥综合征小鼠颌下腺局部注射间充质干细胞(mesenchymal stem cells,MSCs)后的疗效和机制,为临床应用间充质干细胞治疗干燥综合征提供依据。方法选取公认的非肥胖性糖尿病(non-obese diabetic,NOD)小鼠作为干燥综合征模型,颌下腺部位注射5×10~5MSCs,1个月后,检测小鼠唾液流量,观察颌下腺淋巴细胞浸润情况,采用流式细胞术和TUNEL法检测颌下腺细胞凋亡。结果与对照组相比,颌下腺注射MSCs组小鼠唾液流量明显增加,颌下腺浸润淋巴细胞明显减少,颌下腺细胞凋亡明显减少。结论颌下腺局部注射MSCs可以治疗干燥综合征,其机制在于抑制颌下腺细胞凋亡和减少淋巴细胞浸润,为临床应用MSCs治疗干燥综合征提供了依据。     

颌下腺内生物活性物质的研究进展

自从Werle和Roden[1]于1936年在颌下腺中发现激肽释放酶以来,颌下腺的功能便引起了人们的关注。后来,随着神经生长因子(NGF)[2]和表皮生长因子(EGF)[3]相继在颌下腺中被发现,医学上掀起了研究颌下腺内生物活性物质(BAS)的高潮。,几十年来,人们在颌下腺中陆续发现或分离提取出50多种BAS,它们或直接分泌入血,或随唾液进入消化道再由胃肠吸收入血,对多种组织和细胞的生理活动起调节作用。现已证实,啮齿动物颌下腺中的多种BAS主要定位于颗粒曲管(GCT)细胞的分泌颗粒内,而人和其他哺乳动物的颌下腺无GCT细胞,这些物质很可能由纹状管或…     

炎症小体激活参与干燥综合征的实验研究

通过研究炎症小体激活与干燥综合征(Sj?gren’ssyndrome,SS)小鼠组织病理学改变和唾液流量等变化的关系,阐述炎症小体参与SS的作用机制。选用非肥胖型糖尿病(non-obesediabetic,NOD)小鼠作为SS模型,美国癌症研究所(Institute of Cancer Rsearche,ICR)小鼠作为对照;检测和比较SS和对照小鼠唾液流量、颌下腺和肺脏组织病理学表现及炎症小体表达水平;分别注射LPS激活或者注射BAY＿11-7082抑制NOD小鼠炎症小体通路,检测处理后小鼠唾液流量、颌下腺和肺脏组织病理学表现以及炎症小体通路分子表达变化。结果显示,与ICR小鼠相比,NOD小鼠唾液流量减少(P<0.05),颌下腺有淋巴细胞浸润,肺部存在慢性病变,炎症小体通路活化;LPS注射进一步激活SS小鼠体内炎症小体后,唾液流量降低(P<0.001),颌下腺和肺部炎性淋巴细胞浸润增多,血清中IL-1β和IL-18表达增加(均P<0.05);BAY＿11-7082抑制SS小鼠体内炎症小体通路后,唾液流量增加(P<0.05),颌下腺和肺部炎性淋巴细胞浸润减少,血...     

唾液中表皮生长因子来源分析

采用放射免疫测定方法,分别检测了46位受试者的混合唾液、腮腺唾液和颌下腺舌下腺混合唾液内的EGF浓度。结果:腮腺唾液EGF为1743～3472pg/ml,平均2360pg/ml(n=46),比混合唾液EGF高(906～2190pg/ml,平均1604pg/ml,n=46,P<0.01),也比颌下腺舌下腺混合唾液的含量高(427～816pg/ml,平均627pg/ml,n=39,P<0.01)。三组唾液在男女性中的差异无统计学意义。结果说明唾液中EGF主要来源于腮腺,而与以往普遍认为来源于颌下腺的看法不同。     

甲磺酸伊马替尼调控Telocytes对下颌下腺唾液分泌功能的影响

目的探讨甲磺酸伊马替尼调控Telocytes(TCs)对下颌下腺唾液分泌功能的影响。方法采用免疫荧光染色技术对TCs免疫标记物(C-kit/CD117、CD34)进行双标记染色,进而对小鼠下颌下腺内TCs进行定位。采用透射电子显微镜(TEM)显示小鼠下颌下腺内TCs的超微结构及其与周围细胞之间的关系。建立甲磺酸伊马替尼干预模型,实验小鼠分5组:正常组(共24只),用药1周、2周、3周、4周组(共24只),药物以80 mg/(kg·d)每日灌胃,免疫荧光显示,用药前后下颌下腺内TCs的变化;免疫印迹法(Western blotting)观察用药前后CD117、CD34及下颌下腺内α唾液淀粉酶(α-Amy)蛋白表达水平的变化。结果免疫荧光染色显示,TCs广泛分布于下颌下腺结缔组织内,胞体较小有突起(Tps),Tps相互连接成网络样结构,包绕着腺泡及导管,随甲磺酸伊马替尼干预时间延长,用药组Tps构成的网络样结构变稀疏。超微结构显示,Tps呈念珠状,与邻近组织紧密相连,周围可见胞外囊泡。随用药时间的增加Tps减少,TCs胞内囊泡增多,胞内细胞器减少。免疫印迹法显示,用药后CD117、CD34、α-Amy蛋白表达水平相应减少且互为正相关。结论小鼠下颌下腺内存在TCs,甲磺酸伊马替尼对TCs的干预可能会通过影响TCs的结构、免疫表型和细胞间通讯降低下颌下腺唾液分泌功能。     

唾液中表皮生长因子含量测定

采用放射免疫测定方法,分别检测了46位受试者的混合唾液、腮腺唾液和颌下腺舌下腺混合唾液内的EGF浓度。结果:腮腺唾液EGF为1743～3472Pg/ml,平均 2360±422Pg/ml,比混合唾液EGF高(906～2190pg/ml,平均1604±321pg/ml,P<0.01),也比颌下腺舌下腺混合唾液的含量高(427～816pg/ml,平均627±107pg/ml,P<0.O1)。3组唾液在两性中的差异无显著,但40岁以上唾液EGF含量较低,它与40岁以下者之间的差异有显著(P<0.05)。结果说明唾液中EGF主要来源于腮腺,而与以往普遍认为来源于颌下腺的看法不同。     

麻黄汤及拆方对汗液、唾液分泌的影响

目的:中药复方“麻黄汤”是张仲景《伤寒论》中发汗解表的著名方剂。本课题以发汗为主要药理学观察指标,通过研究麻黄汤及拆方对汗液、唾液分泌的影响来探讨麻黄汤发汗的主要药物及物质基础。方法:采用汗腺上皮组织形态观测法、汗液着色法、唾液分泌法等进行研究。结果:(1)麻黄汤及拆方对正常大鼠足跖部汗腺组织形态学变化表明:麻黄汤标准汤及去甘草组发汗效果最好,其余各方也有一定发汗作用。(2)麻黄汤及拆方对正常大鼠足跖部汗腺分泌的影响,各组     

Pituitary adenylate cyclase‐activating polypeptide enhances saliva secretion via direct binding to PACAP receptors of major salivary glands in mice

Xerostomia, or dry mouth, is a common syndrome that is generally treated with artificial saliva; however, no other effective methods have yet been established. Saliva secretion is mainly under the control of the autonomic nervous system. Pituitary adenylate cyclase‐activating polypeptide (PACAP) is recognized as a multifunctional neuropeptide in various organs. In this study, we examined the effect of PACAP on saliva secretion, and detected the distribution of the PACAP type 1 receptor (PAC1R) in major salivary glands, including the parotid, submandibular, and sublingual glands, in 9‐week‐old male C57BL/6 mice. Intranasal administration of PACAP 38 increased the amount of saliva secreted, which was not inhibited by atropine pretreatment. Immunohistochemical analysis showed that PAC1R was distributed in the three major salivary glands. In the parotid and sublingual glands, PAC1R was detected in striated duct cells, whereas in the submandibular gland, a strong PAC1R immunoreaction was detected in tall columnar epithelial cells in the granular ducts (i.e., pillar cells), as well as in some striated duct cells. PACAP significantly increased the concentration of epidermal growth factor in saliva. These results suggest that PACAP directly regulates saliva secretion by controlling the absorption activity in the ducts, and that pillar cells regulate the function of granular epithelial cells in the granular duct, such as the secretion of growth factors into the saliva. Collectively, these results suggest the possibility of PACAP as a new effective treatment of xerostomia. Anat Rec, 299:1293–1299, 2016. © 2016 Wiley Periodicals, Inc.     

鼻咽癌IMRT前后唾液腺功能变化及与受照剂量的关系

目的:探讨鼻咽癌患者调强放射治疗（IMRT）前后唾液腺的功能变化及与受照剂量的关系。方法:选取广西医科大学第一附属医院接受IMRT初治鼻咽癌患者30例为研究对象,在放疗前、放疗后3个月采用99mTcO4-SPECT唾液腺动态显像测定腮腺、颌下腺的时间-放射性曲线（TAC）、最大浓聚率（MAR）和酸刺激最大分泌率（MSR）,研究唾液腺的功能变化及与受照剂量的关系。结果:放疗后3个月出现1~2级口干症状,腮腺、颌下腺TAC曲线主要表现为轻中度受损,口干程度、TAC曲线与唾液腺受照剂量正相关。放疗后3个月较放疗前腮腺MAR、MSR和颌下腺MSR明显减低（P<0.05）,而颌下腺的MAR无减低（P>0.05）,但两组唾液腺的MAR、MSR对比差异无统计学意义（P>0.05）,两组唾液腺照射剂量均合理。结论:鼻咽癌患者IMRT后出现唾液腺摄取和排泄功能轻中度受损,引起1~2级口干,IMRT能够将唾液腺的受照射剂量控制在合理范围内。     

Fluid and protein secretion from ferret submandibular and parotid glands in response to sympathetic nerve stimulation or administration of sympathomimetics.

Electrical stimulation of the sympathetic innervation evoked secretion of submandibular and parotid saliva. By changing the mode of stimulation from a continuous to an intermittent one the fluid response increased and glandular blood flow improved. The volumes from the submandibular glands were larger than those from the parotid glands and further, the protein concentration of submandibular saliva was higher than that of parotid saliva. Adrenaline, isoprenaline and phenylephrine evoked larger fluid responses from submandibular than from parotid glands. However, the fluid response was small compared to the parasympathetic one. Substance P-evoked saliva was used as carrier for protein released by sympathetic nerve stimulation or administration of adrenaline and isoprenaline. In vitro tissues of submandibular and parotid glands responded to adrenaline with a dose-dependent release of protein. Taken together, the analytical pharmacology performed in vivo and in vitro, and including the antagonists phentolamine, dihydroergotamine, propranolol and metoprolol, showed that in submandibular glands, α(α₁)adrenoceptors were predominantly involved in fluid secretion and β(β₁)-adrenoceptors predominantly involved in protein secretion. In parotid glands, fluid secretion seemed solely to depend on α(α₁)-adrenoceptors, while β(β₁)-adrenoceptors seemed almost solely involved in protein secretion.     

Protein secretion in salivary glands of cats in vivo and in vitro in response to vasoactive intestinal peptide

In anaesthetized cats exogenous vasoactive intestinal peptide failed to elicit any secretion of saliva from the submandibular and parotid glands. However, protein release from both glands occurred in response to VIP in the presence of alpha- and beta-adrenoceptor blocking agents and was dose-dependent. This response was revealed by means of a subsequent washout flow of saliva evoked by intravenous injections of methacholine or stimulations of the parasympathetic innervation. The submandibular glands responded to vasoactive intestinal peptide at a lower dose than the parotid glands. In the presence of atropine (but in the absence of adrenoceptor blockers), stimulation of the parasympathetic chorda-lingual nerve, which of itself elicited no secretion of saliva, contributed to the release of protein within the submandibular gland, since the output of protein in response to a subsequent stimulation of the sympathetic innervation was increased. Vasoactive intestinal peptide administered in combination with methacholine or during ongoing parasympathetic nerve-induced salivary secretion revealed positive interactions, particularly with respect to protein release. In-vitro protein release in response to vasoactive intestinal peptide was also demonstrated by perfusing small pieces of the two glands in the presence of muscarinic and adrenoceptor blockers. As in vivo, submandibular tissue responded at a lower concentration of vasoactive intestinal peptide than the parotid tissue. One to two weeks after combined parasympathetic and sympathetic denervation of the parotid glands, the glands were sensitized to vasoactive intestinal peptide when tested in vitro. It is concluded that vasoactive intestinal peptide or a structurally related peptide is a potential transmitter in the parasympathetic control of protein secretion in salivary glands of cats.     

Regulation of Salivary Kallikrein Secretion in the Rat Submandibular Gland

Unstimulated pairs of rat submandibular glands were compared with regard to their wet weight, total protein content and kallikrein activity quantitated by Bz-Arg-OEt-esterase and kallikrein antigenic activity. Paired glands from the same animal were found to be comparable, whereas differences from one animal to another were considerable. One of two paired glands was extirpated and used as control, and the other was subsequently subjected to stimulation. Salivary secretion was induced parasympathomimetically (intraperitoneal injections of pilocarpine; perfusion with acetylcholine and electrical stimulation of the ductal nerve plexus near the gland hilus) or sympathomimetically (cervical sympathetic nerve stimulation with or without administration of α or β-adrenergic blocker; perfusion with epinephrine, norepinephrine or isoproterenol). The effect was studied by measuring the change in total gland kallikrein content and by quantitation of kallikrein in saliva. A small secretion of kallikrein was always observed. However, oc-adrenergic stimulation was 40 and 1500 fold more effective in releasing kallikrein than β-adrenergic and parasympathomimetic stimulation, respectively. Also, significantly more kallikrein was released by β-adrenergic than parasympathomimetic stimulation. Immunohistochemistry confirmed the observed depletion of kallikrein following α-adrenergic stimulation. No alteration in kallikrein localization was observed in stimulated glands.     

Submandibular and parotid salivary secretion after electrolytic lesioning of the brainstem nucleus parvocellularis in the rat

The present study, in consonance with recent anatomical investigations, demonstrates that activation of the nucleus parvocellularis in the rat evokes a potent hypersecretory effect in the submandibular and sublingual (S-S) salivary glands. Furthermore, electrolytic lesioning of this region in conjunction with peripheral removal of the parotid glands is followed by an increase in the number of drinking responses in the presence of dry food. Such prandial drinking behavior is only observed after total impairment of salivation (i.e., removal of the S-S + parotid glands), thus suggesting that the parvocellularis lesion led to a marked deficit in S-S salivary secretion. On the other hand, the activation of the nucleus parvocellularis was seen to have only a slight effect on parotid salivary secretion. Electrolytic lesions to this zone, when associated with peripheral removal of the S-S glands, failed to induce prandiality, suggesting that the parvocellularis nucleus exerted a low level of control over parotid salivary secretion. These results are interpreted as functional proof of the relationship between the parvocellularis reticular formation and the superior salivatory nucleus in the secretion of S-S saliva.     

Mechanism of action of calcitonin on secretion in rat submandibular gland

Calcitonin (CT) was found to reduce the initial flow of pilocarpine-stimulated saliva from the submandibular glands in the rat. Although there was a concomitant increase of the concentration of calcium and protein in the saliva, the calcium/protein ratio was not significantly affected. CT also caused a significant increase of the potassium concentration in submandibular saliva. Both in vivo and in vitro, CT inhibited the production of cyclic AMP (cAMP) both in the absence and in the presence of forskolin. This decrease in intracellular cAMP levels could result in an inhibition of mucus secretion, which would explain the previously observed calcitonin-induced intracellular accumulation of mucus in the submandibular gland acinar cells. CT did not affect the cytoplasmic free Ca2+ concentration (as measured with fura 2) in isolated submandibular acini either in the absence or in the presence of cholinergic or adrenergic agonists. These results indicate that the inhibition of fluid secretion in the submandibular gland by calcitonin must be located distal to changes in [Ca2+]i. It can be concluded that CT affects both mucus and fluid secretion in the submandibular gland, but that only the inhibition of mucus secretion can as yet be explained by an effect at the level of the second messenger.     

Neuronal VIP in salivary glands: distribution and release

Nerves containing vasoactive intestinal peptide (VIP) were observed in salivary glands of rat, cat and man. VIP nerves were numerous in the cat while they were moderate in number in rat and man. The measured concentrations of immunoassayable VIP were in agreement with the immunohistochemical findings. Electrical stimulation of the feline chorda lingual nerve, which stimulates salivary secretion and local blood flow, resulted in a marked elevation of VIP in the venous effluent from the submandibular gland. VIP was not measurable in saliva. Gel permeation chromatography of extracts from cat submandibular gland and from venous plasma collected before and during nervous stimulation revealed one immunoreactive peak with an elution position identical to that of highly purified porcine VIP. The finding of neuronal VIP in salivary glands, its release upon nerve stimulation and its known effect on local blood flow support the view that VIP is a neurotransmitter in the salivary glands.     

Whole, submandibular, and parotid saliva-mediated aggregation of Pseudomonas aeruginosa in cystic fibrosis.

The aggregation of mucoid and nonmucoid Pseudomonas aeruginosa by submandibular, parotid, and whole saliva from patients with cystic fibrosis (CF) and non-CF subjects was investigated. There were significant differences (P less than 0.01) in aggregation of mucoid and nonmucoid variants of P. aeruginosa by submandibular and whole saliva from CF patients and non-CF subjects. However, the differences in the parotid secretion were not as pronounced. Patients with CF who were colonized with P. aeruginosa demonstrated a significantly higher (P less than 0.05) percent aggregation of the mucoid variants by the submandibular secretion and of both mucoid and nonmucoid variants by whole saliva, compared with corresponding secretions from patients with CF not colonized with this pathogen. The parotid saliva aggregation activity was not markedly different for the two groups with CF. From patients with CF, whole saliva demonstrated a higher percent P. aeruginosa aggregation than did the submandibular saliva. In non-CF subjects, however, the percent aggregation of P. aeruginosa by submandibular saliva was higher than that by whole saliva. Our results indicate that the sero-mucous products of the submandibular gland have a more significant role in P. aeruginosa aggregation than the serous secreting parotid cells and that the submandibular secretion is possibly responsible for the differences in oral colonization by this pathogen in subjects with and without CF.     

Modulation of carbachol-induced Cl(-) currents and fluid secretion by isoproterenol in rat submandibular acinar cells.

The effects of muscarinic and beta-adrenergic agonists on Cl(-) currents in acinar cells were investigated to clarify their role in the regulation of fluid secretion in rat perfused submandibular glands. Additions of isoproterenol (IPR) at 10(-8) to 10(-6) M and 8-(4-chlorophenylthio)-cyclic AMP (CPT-cAMP) at 10(-3) M to the perfusate suppressed carbachol (CCh; 10(-6) M)-induced fluid secretion. IPR and CPT-cAMP also diminished CCh-induced oscillatory Cl(-) current and increased CCh-stimulated non-oscillatory Cl(-) current. Propranolol blocked the effect of IPR on fluid secretion. IPR did not modulate the CCh-induced increase in intracellular concentration of calcium ions and intracellular pH in isolated cells. Propranolol blocked IPR-induced changes in Cl(- )currents, while propranolol itself increased CCh-induced K(+) current and reduced CCh-induced oscillatory Cl(-) current. Increasing external osmolarity with 50 mM sucrose abolished IPR-enhanced non-oscillatory Cl(-) current. Neither CCh-induced oscillatory Cl(-) current nor IPR-induced suppression of the oscillatory Cl(-) current was influenced by the hypertonicity. Perfusion of the gland with the hypertonic solution did not affect the IPR-induced suppression of fluid secretion. These results suggest that IPR induces the suppression of CCh-induced oscillatory Cl(-) current and potentiation of the non-oscillatory Cl(-) current via an increase in cyclic AMP level, and that suppression of the oscillatory Cl(-) current by IPR may contribute to the inhibition of fluid secretion from submandibular glands.