miR-375在变应性鼻炎小鼠鼻黏膜上皮细胞凋亡中的调控作用

目的探讨miR-375在变应性鼻炎小鼠鼻黏膜上皮细胞凋亡和炎症反应中的调控作用。方法运用卵清蛋白(OVA)致敏的小鼠变应性鼻炎模型,使用实时定量PCR(qRT-PCR)、蛋白质印迹试验(Western Blot)、酶联免疫吸附试验(ELISA)、免疫组织化学检测鼻黏膜上皮细胞内miR-375、JAK2、细胞凋亡相关蛋白(JAK2蛋白,裂解的蛋白酶3(Cleaved caspase 3),聚[ADP-核糖]聚合酶裂解酶(Cleaved PARP),蛋白酶3(Caspase 3),聚[ADP-核糖]聚合酶(PARP),p-STAT3蛋白,STAT3蛋白和β肌动蛋白(β-actin))和血浆IL-6、TNF-α、IL-10的表达水平。结果miR-375在变应性鼻炎小鼠的鼻黏膜上皮细胞中表达降低,而JAK2表达增高;JAK2蛋白、p-STAT3蛋白和裂解的蛋白酶3均在OVA组表达增高;给OVA致敏的变应性鼻炎小鼠注射miR-375模拟物可以导致血清IL-6、TNF-α的分泌下降,而IL-10分泌增加,该作用可以被带有过表达JAK2的腺病毒感染后而减弱。结论miR-375/JAK2调控通路存在于变应性鼻炎鼻黏膜上皮细胞中,并通过JAK2/STAT3信号通路调控细胞的凋亡和炎症反应,miR-375在变应性鼻炎的病程中有保护性机制。     

小鼠哮喘模型气道上皮内肥大细胞数量增多

目的:观察小鼠哮喘模型气道卜皮内肥大细胞的数量,进一步阐明肥大细胞在鸡卵清蛋白(OVA)诱导的小鼠哮喘模型中的作用。方法:OVA诱导建立小鼠哮喘模型,H-E染色病理切片观察肺部感染情况,甲苯胺蓝染色法观察气道上皮内肥大细胞数量的变化。结果:哮喘模型组小鼠肺部炎症明显,在正常组和对照组小鼠气道上皮内没有发现肥大细胞,但在模型组小鼠气道上皮内肥大细胞的数量随OVA的激发次数增加而增多。结论:在OVA诱导的小鼠哮喘模型气道上皮内肥大细胞随着激发次数的增加而增多。气道上皮内肥大细胞在OVA诱导的哮喘模型起重要作用。     

重组E.coli. LLO/OVA明显增强小鼠CD11c细胞活性及抗肿瘤免疫

目的 探讨重组E.coli. LLO/OVA 诱导C57BL/6小鼠机体免疫的途径,观察其免疫后小鼠机体抑制B16-OVA黑色素瘤的效果.方法 磁珠分离E.coli. LLO/OVA及E.coli. OVA免疫后小鼠脾脏CD11c、CD4 和CD8 T细胞并检测CD11c细胞诱导同源CD4 和CD8 T增殖水平和细胞因子分泌程度;流式细胞检测小鼠脾脏内肿瘤抗原OVA257-264 SIINFEKL特异的细胞毒T细胞含量.比较两种E.coli. 免疫后,恶性黑色素瘤B16-OVA在小鼠肺内形成瘤结节的数量.结果 与E.coli. OVA相比, E.coli. LLO/OVA免疫后小鼠脾脏CD11c细胞诱导同源CD4 T细胞增殖作用增强、IL-2分泌增高;同时诱导CD8 T细胞增殖和IFN-γ分泌的作用也明显增强;OVA257-264 SIINFEKL特异的CD8 T细胞含量也明显增高;小鼠肺内形成B16-OVA瘤结节平均数明显减少. 结论 E.coli. LLO/OVA有效地诱导小鼠CD11c细胞活化,增强其对CD4 T细胞增殖和IL-2分泌以及对CD8 T细胞增殖、IFN-γ分泌的作用,诱导了更多的OVA特异的CD8 T细胞,使机体产生了更强的抗肿瘤免疫.     

Stable dry powder inhaler formulation of tranilast attenuated antigen-evoked airway inflammation in rats

Tranilast (TL) has been clinically used for the treatment of airway inflammatory diseases, although the clinical use of TL is limited because of its poor solubility and systemic side effects. To overcome these drawbacks, a novel respirable powder of TL (CSD/TL-RP) for inhalation therapy was developed using nanocrystal solid dispersion of TL (CSD/TL). Stability study on CSD/TL-RP was carried out with a focus on inhalation performance. Even after 6 months of storage at room temperature, there were no significant morphological changes in micronized particles on the surface of carrier particles as compared with that before storage. Cascade impactor analyses on CSD/TL-RP demonstrated high inhalation performance with emitted dose and fine particle fraction (FPF) of ca. 98% and 60%, respectively. Long-term storage of CSD/TL-RP resulted in only a slight decrease in FPF value (ca. 54%). Inhaled CSD/TL-RP could attenuate antigen-induced inflammatory events in rats, as evidenced by marked reduction of granulocytes in bronchoalveolar lavage fluid and inflammatory biomarkers such as eosinophil peroxidase, myeloperoxidase, and lactate dehydrogenase. These findings were consistent with decreased expression levels of mRNAs for nuclear factor-kappa B and cyclooxygenase-2, typical inflammatory mediators. Given these findings, inhalable TL formulation might be an interesting alternative to oral therapy for the treatment of asthma and other airway inflammatory diseases with sufficient dispersing stability.     

内脏高敏感状态下食管酸灌注诱导的鼠脊髓背角c-Fos基因表达的研究

目的 观察在内脏高敏感状态下,食管酸灌注诱导的大鼠脊髓背角Fos 蛋白表达的表达,初步探索食管内脏感觉过敏在脊髓水平敏感化的分子机制.方法 采用腹腔注射鸡卵清蛋白基础致敏联合食管酸灌注的方法,建立内脏高敏感性-食管化学刺激大鼠模型;采用免疫组织化学方法和显微图像分析技术研究,在生理条件、内脏高敏感状态下进行食管酸灌注时Fos蛋白激活模式的差异.结果 模型组大鼠双侧脊髓背角内有大量的Fos样免疫反应 (FLI) 阳性神经元,集中分布于背角Ⅰ～Ⅱ、Ⅴ～Ⅶ层,其FLI阳性神经元数量和平均光密度值较单纯食管酸灌注组和单纯,OVA(ovalbumin)致敏组明显增加,差异有统计学意义(P＜0.05).单纯酸灌注组和单纯OVA致敏组在背角Ⅰ～Ⅱ、Ⅴ～Ⅵ、Ⅹ层的FLI阳性神经元数量均较生理盐水对照组显著增加,差异有统计学意义(P＜0.01).单纯酸灌注组在背角Ⅰ～Ⅱ层和Ⅲ～Ⅳ层FLI阳性细胞数较单纯OVA致敏组显著增加,差异有统计学意义(P＜0.05).结论 腹腔注射鸡卵清蛋白基础致敏,对食管酸灌注诱导的脊髓背角内Fos蛋白表达有活化作用,c-Fos过度表达的阳性神经元的兴奋性增加和神经可塑性改变,促进了内脏高敏感性在脊髓水平敏感化的形成和维持.     

环孢素A气雾给药对豚鼠气道高反应性和炎症的作用

目的:评价环孢素A气雾给药对豚鼠哮喘模型的药效.方法:用乙酰胆碱(ACh)或组胺诱导抗原攻击后的致敏豚鼠气道阻力PC_(200)、支气管肺泡灌洗液(BALF)和肺组织切片中的嗜酸性粒细胞(EOS)变化观察环孢素A气雾给药后的抗气道高反应性和炎症作用.结果:环孢素A 10 g·L~(-1)、20 g·L~(-1)气雾给药和地塞米松(0.5mg·kg~(-1),ip)增加PC_(200)值,能预防ACh或组胺引起的气道高反应性,环孢素A 5 g·L~(-1)对组胺引起的气道高反应性也有作用,对ACh不显著.环孢素A 10 g·L~(-1)、20 g·L~(-1)气雾给药能明显减少BALF中的EOS浸润.与溶媒组比较,地塞米松0.5mg·kg~(-1)增加了BALF中的中性粒细胞数目,与三组环孢素A比较有显著差异.在肺组织学研究中,环孢素A 20 g·L~(-1)和地塞米松0.5 mg·kg~(-1)可抑制支气管和细支气管上皮和上皮表面结缔组织的EOS浸润.结论:环孢素A气雾吸入给药能明显对抗致敏豚鼠气道高反应性和炎症反应,为其治疗哮喘提供了一个可选择的给药途径.     

Inhibitory effects of cryptoporus polysaccharide on airway constriction, eosinophil release, and chemotaxis in guinea pigs

AIM: To study effects of cryptoporus polysaccharide (CP) on antigen-induced bronchoconstriction, eosinophil peroxidase (EPO) release in vivo, and on platelet activating factor (PAF)-induced eosinophil chemotaxis in vitro in guinea pig. METHODS: The asthma model of guinea pig was formed with ovalbumin (OVA). The changes of lung resistance (RL) and dynamic lung compliance (Cdyn), EPO level in bronchoalveolar lavage fluids (BALF) and eosino- phil migration were determined. RESUL…     

Artificial Tears Inhibit the Development of Experimental Immune-mediated Blepharoconjunctivitis

Purpose To investigate whether treatment with artificial tears inhibits the development of experimental immune-mediated blepharoconjunctivitis (EC).Methods Brown Norway rats were immunized with ovalbumin (OVA) or ragweed (RW) emulsified in complete Freunds adjuvant. Fourteen days after immunization, the rats were challenged with the same antigen (Ag) in eye drops. Treated rats were administered artificial tears by eye drops immediately after, 15min after, or 30min after the Ag challenge. Treatment doses of 2, 4, or 8 drops per eye were evaluated. Twenty-four hours after the Ag challenge, the rats were killed and their eyes were harvested for histological studies.Results Treatment with artificial tears immediately after and 15min after challenge with partially insoluble RW Ag suppressed infiltration of inflammatory cells into the conjunctiva. Inhibition was not observed at any time following challenge with OVA Ag, which is a soluble protein. The treatment dose of artificial tears administered did not affect the extent of inhibition of EC following challenge with either Ag.Conclusions Treatment with artificial tears by eye drops inhibited the development of EC induced by the partially insoluble RW Ag when administered within 15min of the Ag challenge. Jpn J Ophthalmol 2004;48:530–534 © Japanese Ophthalmological Society 2004     

Oral-Antigen Delivery via a Water-in-Oil Emulsion System Modulates the Balance of the Th1/Th2 Type Response in Oral Tolerance

Purpose. To evaluate the ability of a water-in-oil (W/O) emulsion containing ovalbumin (OVA), a model antigen, to induce oral tolerance and to elucidate the mechanism for the induction of oral tolerance by the emulsion system. Methods. A W/O emulsion containing OVA was prepared and evaluated its ability to induce oral tolerance in mice. Also, the Th1/Th2 balance in the mice tolerized was investigated in terms of the ratios of anti-OVA IgG2a titer to anti-OVA IgG1 titer (IgG2a/IgG1 ratios) and cytokine profiles. Results. Anti-OVA total IgG antibody titer of mice administered OVA in saline was approximately 3.5-fold higher than that of the mice administered OVA in W/O emulsion at a dose of 0.1 mg/mouse/day. Similar total IgG responses were observed between the above two at a dose of 1 mg/mouse/day. The IgG2a/IgG1 ratios decreased as the dose of OVA in W/O emulsion, but not in saline, increased at doses of 0, 0.1, and 1 mg/mouse/day. Interferon- secretion of PLN cells from the mice administered OVA in W/O emulsion decreased, whereas their interleukin-4 secretion remained high. Although interferon- secretion for the mice administered OVA in saline decreased, interleukin-4 secretion did not change. Conclusions. The present study suggests that oral delivery of OVA via the W/O emulsion system may more efficiently enhance the induction of Th2-dominated imbalance than that of OVA in saline.