补气固表制剂对变应性鼻炎鼻黏膜细胞凋亡指数的双向调节作用

目的：探讨补气固表制剂对变应性鼻炎鼻黏膜细胞凋亡指数的双向调节作用。方法：制作变应性鼻炎豚鼠模型，观察黏膜上皮细胞，黏膜下腺体细胞凋亡指数的变化和细胞凋亡与嗜酸细胞数量的关系，观察黏膜上皮细胞、黏膜下腺体细胞凋亡指数比值对补气固表制剂的反应，并与对照组比较。结果：变应性鼻炎模型组上述指标均异常，与正常对照组相比差异有统计学意义（P〈0．01）。补气固表制剂干预后恢复正常。与正常对照组相比差异无统计学意义（P〉0．05）。结论：变应性鼻炎鼻黏膜细胞的凋亡具有双重性，其凋亡特性与细胞种类相关，并与嗜酸细胞的数量有关，补气固表制剂能双向调节凋亡指数，保持鼻黏膜的生理稳态。有效地治疗变应性鼻炎。     

玉屏风散对变应性鼻炎大鼠鼻黏膜NF-κBp65、GATA-3表达的影响

目的通过研究变应性鼻炎大鼠鼻黏膜NF-κBp65、GATA-3表达的影响,探讨玉屏风散治疗变应性鼻炎的可能机制。方法 60只SD大鼠按体质量随机分为6组,每组10只,分别为正常组、模型组、治疗组(玉屏风散低、中、高剂量组)、对照组。除正常组外,其余的大鼠均建立卵清蛋白诱导的变应性鼻炎模型。分组给药干预1周。应用HE染色检测鼻黏膜组织结构变化,应用western-blot及Realtime PCR技术检测NF-κBp65、GATA-3在鼻黏膜组织中的表达。结果模型组鼻组织病理学表现以组织重塑、上皮细胞脱落、杯状细胞化生、组织水肿、大量嗜酸性粒细胞浸润为主;治疗组的表现以鼻黏膜存在组织重塑表现,鼻黏膜上皮细胞脱落、组织间质轻度水肿、固有层可见少量嗜酸性粒细胞浸润,两组比较,有统计学差异(P<0.05)。模型组鼻黏膜中NF-κBp65、GATA-3蛋白及其mRNA呈高表达变化;给予玉屏风散治疗后,NF-κBp65、GATA-3蛋白及其mRNA表达下降,经统计学比较,有显著性差异(P<0.05)。结论变应性鼻炎鼻黏膜存在组织重塑,玉屏风散可能通过下调NF-κBp65、GATA-3表达,发挥防治变应性鼻炎的效应。     

变应性鼻炎大鼠鼻黏膜Bcl-2蛋白表达与细胞凋亡的研究

目的探讨变应性鼻炎(AR)大鼠鼻黏膜Bcl-2蛋白的表达与细胞凋亡之间的关系,进一步了解变应性鼻炎的发病机制。方法选健康SD大鼠20只,雌雄不限,随机分为实验组和对照组,各10只。实验组以卵清蛋白腹腔注射致敏,继之鼻局部激发建立变应性鼻炎大鼠模型。取实验组与对照组动物鼻呼吸区黏膜,行Bcl-2蛋白免疫组织化学染色,以细胞凋亡末端脱氧核苷酸转移酶介导的dUTP缺口末端标记(TUNEL)法检测鼻黏膜细胞凋亡状况,以图像分析软件分析结果。结果变应性鼻炎大鼠鼻黏膜Bcl-2蛋白主要表达于细胞胞质中,包括鼻黏膜腺体细胞、上皮细胞,固有层中的淋巴细胞,鼻分泌物中可见大量呈强阳性反应的嗜酸性粒细胞,实验组Bcl-2蛋白表达水平显著高于对照组。凋亡细胞散在于鼻黏膜上皮层和固有层,实验组更多见于上皮层,实验组凋亡指数与对照组相比下调。结论变应性鼻炎大鼠鼻黏膜细胞凋亡过程受抑可能是变应性鼻炎鼻分泌物增多及上皮破坏的机制之一。     

水通道蛋白3和水通道蛋白9在结肠黏膜的表达及意义

目的 探讨结肠黏膜水通道蛋白3(aquaporin,AQP3)和水通道蛋白9(aquaporin,AQP9)的表达与分布情况,并探讨其与便秘的关系.方法 先用免疫组织化学技术检测45例功能性便秘(functional constipation,FC)患者(试验组)和32例非便秘患者(对照组)结肠黏膜AQP3和AQP9的分布情况;然后用Western bloting半定量检测AQP3和AQP9,以β-actin作内参蛋白,两者与相应β-actin灰度值之比即为AQP3和AQP9的相对含量,将灰度比值作统计学分析.结果 AQP3主要分布在肠黏膜上皮细胞的基底膜,AQP9主要分布在黏膜腔面杯状细胞.右半结肠试验组AQP3含量高于对照组(P＜0.05),AQP9差异无统计学意义(P＞0.05),而左半结肠试验组AQP9低于对照组(P＜0.05),AQP3差异无统计学意义(P＞0.05).结论 AQP3在右半结肠高表达,而AQP9在左半结肠高表达;功能性便秘患者右半结肠AQP3增加,而左半结肠AQP9降低;AQP3和AQP9含量的增加或减少可能与便秘的发生发展有一定关系.     

TLR5在变应性鼻炎患者鼻黏膜组织中的表达

目的：检测变应性鼻炎患者鼻黏膜组织中TLR5的表达。方法采用免疫组织化学SABC法检测30例变应性鼻炎患者（试验组）和25例非变应性鼻炎患者（对照组）鼻黏膜组织中TLR5的表达。结果 TLR5在试验组的阳性表达率（83.3%）明显高于对照组（8.0%）,χ^2=30.965,P＜0.05。结论 TLR5在变应性鼻炎患者鼻黏膜组织中高表达。     

变应性鼻炎大鼠鼻黏膜中趋化因子CC的表达

目的探讨大鼠鼻粘膜中趋化因子CC含量与变应性鼻炎（allergicrhinitis）发生的关系。方法提取22只变应性鼻炎组大鼠及对照组鼻黏膜中的总RNA,设计趋化因子CC和内参GAPDHPCR扩增引物,利用SYBRGreen实时荧光定量PCR法检测趋化因子CC及内参的表达水平。结果变应性鼻炎组大鼠鼻黏膜组织中的趋化因子CC的表达量显著高于对照组。结论趋化因子CC可能在变应性鼻炎的发病过程中发挥重要作用。     

蛋白激酶C、Bcl-2及Bax在变应性鼻炎鼻黏膜中的表达及意义

目的探讨变应性鼻炎（AR）患者鼻黏膜蛋白激酶C（PKC）、抗凋亡基因Bcl-2及促凋亡基因Bax蛋白的表达水平,以进一步了解AR的发病机制。方法应用免疫组织化学的方法检测26例AR患者（AR组）鼻黏膜及10例下鼻甲组织中（对照组）PKC、Bcl-2及Bax的表达。结果PKC、Bcl-2及Bax在2组中均有表达,AR组中PKC主要表达在嗜酸粒细胞、血管内皮细胞及腺体细胞,Bcl-2蛋白主要表达于鼻黏膜腺体细胞和上皮层,与对照组相比差异有统计学意义（P〈0.01）。Bax蛋白与Bcl-2蛋白表达部位一致,2组表达水平差异无统计学意义（P〉0.05）。结论AR鼻黏膜蛋白激酶C、抗凋亡基因Bcl-2蛋白表达上调,是AR嗜酸粒细胞增多、鼻分泌物增多及鼻黏膜上皮破坏的机制之一。     

复方鹅芪鼻腔冲洗液治疗变应性鼻炎的效果研究

目的探讨变应性鼻炎患者采用复方鹅芪鼻腔冲洗液治疗的临床效果。方法收集2009年10月～2012年6月来我院进行治疗的变应性鼻炎患者124例,按照治疗方法分为观察组和对照组,每组62例。观察组采用复方鹅芪鼻腔冲洗液治疗;对照组采用生理盐水,3周为1个疗程。两组患者的临床效果及鼻黏膜纤毛清除时间的比较均使用统计学方法进行分析。结果两组患者治疗后的各项症状评分、体征评分、病情评分及鼻黏膜纤毛清除时间与治疗前比较均明显降低;观察组治疗后的症状评分、体征评分、病情评分及鼻黏膜清除时间与对照组治疗后比较明显较低;观察组的总有效率明显高于对照组,两组间差异具有统计学意义(P0.05)。结论变应性鼻炎患者采用复方鹅芪鼻腔冲洗液进行治疗临床效果较好,能够对鼻黏膜上皮的修复起到促进的作用。     

鼻炎通窍喷雾剂对变应性鼻炎豚鼠血清IL-4含量和鼻黏膜Bax蛋白表达的影响

目的:通过研究鼻炎通窍喷雾剂对变应性鼻炎(Allergic rhinitis,AR)模型豚鼠血清中白细胞介素4(Interleukin-4,IL-4)含量和鼻黏膜组织中Bax蛋白表达的影响,探讨鼻炎通窍喷雾剂治疗变应性鼻炎的作用机制。方法:用10%甲苯-2,4二异氰酸酯(10%toluene-2,4-diisocyanate,TDI)橄榄油溶液建立豚鼠变应性鼻炎模型;用酶联免疫吸附法(ELISA)定量测定各组豚鼠血清中IL-4的含量,免疫组化法测定鼻黏膜组织中Bax蛋白的表达。结果:实验组豚鼠血清的IL-4表达水平显著低于模型对照组(p0.01);实验组豚鼠鼻黏膜中Bax蛋白的表达水平显著高于模型对照组(P0.01)。结论:鼻炎通窍喷雾剂对变应性鼻炎的作用机制与调节免疫功能、促进炎细胞凋亡有关。     

硝酸铈对豚鼠变应性鼻炎鼻黏膜的影响

目的探讨1%硝酸铈溶液对豚鼠变应性鼻炎鼻黏膜的影响。方法健康豚鼠39只随机分成正常对照组,变应性鼻炎组：用2,4-二异氰酸甲苯酯（2,4-Toluene-Dissocya-nate,TDI）作致敏原,经鼻腔滴入建立变应性鼻炎模型。治疗期间变应性鼻炎组（n=30）动物随机分成3组：阳性对照组（n=10）、药物治疗对照组（n=10）、硝酸铈治疗组（n=10）,正常对照组（n=9）设为阴性对照组,治疗15d。观察症状体征、鼻分泌物涂片、鼻黏膜组胺含量和光镜和电镜下鼻黏膜组织形态学的变化。结果阳性对照组出现鼻痒、喷嚏、流清涕,鼻分泌物嗜酸性粒细胞增多,鼻黏膜组胺含量增多,嗜酸性粒细胞浸润,鼻黏膜细胞超微结构改变。与阳性对照组相比,硝酸铈治疗组上述变化明显减轻。结论 1%硝酸铈溶液能有效控制豚鼠变应性鼻炎的症状体征,使鼻黏膜嗜酸性粒细胞减少、组胺含量减低。     

Interactions between eotaxin and interleukin-5 in the chemotaxis of primed and non-primed human eosinophils

This study was designed to understand the relationship between interleukin-5 and eotaxin in modulating the chemotaxis of eosinophils obtained from healthy subjects and subjects with allergic rhinitis. Chemotaxis of eosinophils from patients with allergic rhinitis toward interleukin-5 (0.25 ng/ml) was 78% higher than that of healthy subjects. Incubation of eosinophils with eotaxin (100 ng/ml) did not change the interleukin-5-induced chemotaxis of eosinophils from healthy subjects, but it reversed the enhanced chemotaxis seen in eosinophils from allergic patients. Chemotaxis of eosinophils from patients with allergic rhinitis toward eotaxin (100 ng/ml) was 65% higher than that of eosinophils from healthy subjects. Incubation of eosinophils with interleukin-5 (100 ng/ml) significantly increased the eotaxin-induced chemotaxis in both subject groups, but such increases were markedly higher for cells from patients with allergic rhinitis. Our finding that eotaxin inhibits the enhanced eosinophil chemotaxis toward interleukin-5 in primed cells suggests that this chemokine may downregulate eosinophil accumulation in the nasal mucosa of allergic patients.     

白介素-1受体拮抗剂对豚鼠变应性鼻炎的治疗作用

目的：明确白介素-1受体拮抗剂对变应性鼻炎的治疗作用。方法：采用甲苯-2,4-二异氰酸酯（TDI toluene-2,4-diisocyanate)致敏豚鼠,制作变应性鼻炎模型,观察给予白介素-1受体拮抗剂（IL-1ra）处理前后症状和体征的变化,用组织病理学方法观察鼻粘膜的改变,测量鼻粘膜中组胺、血中IgE的含量。结果：用IL-1ra治疗后,变应性鼻炎症状和体征明显改善,HE染色显示,鼻粘膜水肿及炎性细胞浸润,随IL-1ra浓度的提高而改善,鼻粘膜中组胺、血中IgE的含量下降,随白介素-1拮抗剂浓度升高而下降。特别是高剂量组,各指标好于其它各治疗组,阳性对照组也有鼻粘膜水肿,及炎性细胞浸润。结论：局部使用IL-1ra拮抗剂能治疗变应性鼻炎。     

布地奈德对变应性鼻炎豚鼠鼻粘膜嗜酸性粒细胞及组胺的影响

目的：制备变应性鼻炎动物模型,评价布地奈德对变应性鼻炎嗜酸性粒细胞和组胺的影响.方法：将豚鼠随机分成自然对照组、变应性鼻炎组和布地奈德治疗组,采用甲苯-2,4-二异氰酸酯对豚鼠鼻腔局部致敏激发制备变应性鼻炎模型,通过观察动物临床症状、鼻粘膜病理切片和鼻粘膜组织中组胺含量等指标来评价药物的药效.结果：变应性鼻炎组鼻粘膜组织中嗜酸细胞浸润和组胺含量均显著高于自然对照组（P＜0.01或P＜0.05）;经治疗后,动物的嗜酸细胞浸润和组织中组胺含量均明显地降低（P＜0.01）.在临床症状指标上,变应性鼻炎组和药物治疗组评分均明显高于自然对照组（P＜0.01）,但变应性鼻炎组和药物治疗组间无显著差异（P＞0.05）.结论：布地奈德能有效地降低变应性鼻炎豚鼠鼻粘膜组织中嗜酸细胞浸润和组胺的含量,但未能改善动物的鼻部临床症状,这可能与造模的方法有关.     

Effects of lentivirus-mediated CCR3 RNA interference on the function of mast cells of allergic rhinitis in mice

The CC chemokine receptor 3 (CCR3) expressed by eosinophils, mast cells and Th2 cells is closely related to allergic diseases. The objective of this study was to explore whether silencing of CCR3 with short hairpin RNAs (shRNAs) delivered by a lentiviral vector could impact the function of mast cells in a murine model of allergic rhinitis (AR) in vivo. The murine model of allergic rhinitis (AR) inducing by ovalbumin (OVA) was constructed, and the BALB/c mice were divided into normal control group, AR group, controlshRNA treated group and lentiviral CCR3-shRNA treated group. The recombinant lentivirus vectors which express a short hairpin RNA (shRNA) targeting the CCR3 were dropped into the nasal cavity of OVA-sensitized mice before the challenges. Real-time fluorescence quantitative PCR and western blotting were performed to observe inhibitory effect of CCR3 gene. Nasal symptoms of mice and OVA-specific IgE in each group were assessed. Concentrations of histamine, tryptase and Prostaglandin D2 (PGD2) in bone marrow, peripheral blood and nasal mucosa were analyzed. Furthermore, histological analysis and electron microscopy analysis were applied to detect the histology changes of nasal mucosa and the infiltration of mast cells in nasal mucosa. The results showed that administration of CCR3shRNA could effectively inhibit the expression of the CCR3 gene in bone marrow, peripheral blood and nasal mucosa, which reduced the nasal symptoms, the level of OVA-specific IgE, the inflammatory cells and mast cells infiltration into nasal cavity, and relieved the histopathological changes of nasal mucosa. In addition, intervention of CCR3shRNA could reduce the levels of the histamine, tryptase and PGD2 in bone marrow, peripheral blood and nasal mucosa. These results suggest that inhibition of CCR3 gene expression by shRNAs lentiviral vectors can effectively attenuate migration, infiltration and degranulation of mast cells in local tissues and alleviate the inflammation of allergic rhinitis mice.     

五龙颗粒对变应性鼻炎模型大鼠鼻黏膜IL-3 mRNA表达的影响研究

目的:研究五龙颗粒对变应性鼻炎模型大鼠白细胞介素-3(IL-3)mRNA表达的影响,探讨其治疗变应性鼻炎的机制。方法:以卵清蛋白为变应原复制变应性鼻炎模型大鼠,随机分为模型(AR)组、五龙颗粒组和息斯敏组。取3组和健康大鼠(空白组)血清进行组胺检测;取大鼠鼻黏膜作苏木素-伊红染色,并采用逆转录-聚合酶链反应(RT-PCR)检测法对各组大鼠鼻黏膜组织中IL-3mRNA的表达水平进行相对定量比较。结果:IL-3mRNA在4组大鼠鼻黏膜中均有表达,其中AR组的表达水平为11.85±0.82,显著高于空白组(P<0.01);与AR组比较,五龙颗粒及息斯敏组大鼠鼻黏膜IL-3mRNA的表达显著下降(P<0.01)。结论:五龙颗粒能降低模型大鼠鼻黏膜IL-3mRNA的表达水平,可为临床合理用药提供理论基础。     

敏停喷鼻剂对TDI致豚鼠过敏性鼻炎模型的影响

目的研究敏停喷鼻剂对豚鼠过敏性鼻炎模型的影响。方法用TDI致豚鼠过敏性鼻炎造模型，观察敏停喷鼻剂对过敏性鼻炎模型的鼻部过敏症状以及鼻黏膜病理组织形态学的影响。结果敏停喷鼻剂能明显减轻豚鼠过敏性鼻炎模型鼻部过敏症状以及鼻黏膜上皮炎细胞的浸润。结论敏停喷鼻剂对过敏性鼻炎可能具有较好的疗效。     

盐酸左旋咪唑经鼻腔给药治疗小鼠变应性鼻炎的药效学研究

目的:考察盐酸左旋咪唑经鼻腔给药治疗小鼠变应性鼻炎的疗效,为治疗变应性鼻炎寻找新的药物。方法:建立小鼠变应性鼻炎的模型,以布地奈德为阳性对照药,以生理盐水为阴性对照药,经鼻腔给予盐酸左旋咪唑后,观察变应性鼻炎小鼠鼻粘膜的水肿、嗜酸性粒细胞浸润IL-12的表达情况。结果:阳性对照药物雷诺考特与盐酸左旋咪唑都能改善组织水肿、显著抑制嗜酸细胞的浸润并有效地提高组织中IL-12的生成表达;但布地奈德抑制嗜酸细胞效果比盐酸左旋咪唑好,而其促IL-12表达不如盐酸左旋咪唑。结论:盐酸左旋咪唑经鼻腔给药可能对变应性鼻炎具有一定的治疗效果,其机理与布地奈德不一样,有可能成为新的变应性鼻炎的治疗药。     

Prophylactic effects of the histamine H1 receptor antagonist epinastine and the dual thromboxane A2 receptor and chemoattractant receptor-homologous molecule expressed on Th2 cells antagonist ramatroban on allergic rhinitis model in mice

The prophylactic use of anti-allergic drugs has been proposed to be effective in the treatment of seasonal allergic rhinitis in humans. However, there is little information regarding the prophylactic effect of thromboxane A(2) (TXA(2)) receptor antagonist on allergic rhinitis. Recent studies revealed that a TXA(2) receptor antagonist ramatroban could block the prostaglandin D(2) (PGD(2)) receptor and chemoattractant receptor-homologous molecule expressed on Th2 cells (CRTH2). In the present study, we investigated the prophylactic effects of the histamine H(1) receptor antagonist epinastine and the TXA(2) receptor antagonist ramatroban and seratrodast on mouse models of allergic rhinitis. Female BALB/c mice were sensitized by an intraperitoneal injection of ovalbumin and alum on days 0, 5, 14 and 21. Seven days later, mice were sensitized by intranasal application of ovalbumin thrice a week. Drugs were administered once a day from day 22. The severity of allergic rhinitis was assessed by determining the extent of 2 nasal allergic symptoms (sneezing and nasal rubbing). Histamine sensitivity and eosinophil infiltration into the nasal mucosa were also determined. Epinastine and ramatroban significantly reduced nasal symptoms and the number of eosinophils in the nasal mucosa. Seratrodast showed no effect on nasal symptoms and eosinophil infiltration into the nasal mucosa. In addition, histamine sensitivity was reduced by epinastine and ramatroban. These results indicate that epinastine and ramatroban induce the prophylactic effect on allergic rhinitis.