富半胱氨酸61在哮喘小鼠气道上皮细胞中表达的研究

目的 探讨富半胱氨酸61(cyr61)在哮喘小鼠中的表达及氨茶碱对其影响. 方法 40只雌性BALA/c小鼠随机分为哮喘组、氨茶碱干预组、哮喘对照组及正常对照组,每组10只. 哮喘组通过卵清蛋白(OVA)致敏和激发的方法 制作哮喘小鼠模型. 氨茶碱干预组在制作哮喘模型同时,于每次激发后尾静脉注射氨茶碱干预. 哮喘对照组在制作哮喘模型同时,给予0.9%氯化钠溶液替代氨茶碱干预. 瑞特 吉姆萨染色分类计数支气管肺泡灌洗液(BALF)中细胞数,免疫组织化学技术检测小鼠支气管上皮组织cyr61表达,酶联免疫吸附测定(ELISA)检测BALF中白细胞介素4(IL-4)含量. 结果 哮喘组小鼠气道上皮细胞cyr61蛋白表达明显高于正常组小鼠(P<0.05),氨茶碱干预组小鼠气道上皮细胞中cyr61蛋白表达较哮喘组明显降低(P<0.05),但仍高于正常组(P<0.05). cyr61蛋白表达与BALF中有核细胞总数、嗜酸性粒细胞、淋巴细胞以及IL-4呈正相关(P<0.05). 结论 cyr61蛋白可能参与了哮喘的发病过程,其异常表达可能与气道炎症有关.     

气道上皮在哮喘发生发展中的作用及其机制研究

支气管哮喘是由多种细胞、细胞因子和炎症介质引起的以呼吸道高反应性为特征的慢性炎症性疾病.气道上皮可能成为有价值的哮喘治疗靶标,本文综述气道上皮在哮喘发生发展中的作用及相关机制研究进展.     

兰尼碱受体mRNA在哮喘豚鼠气道平滑肌细胞中表达的改变

目的明确兰尼碱受体(RyR)与哮喘发病的关系,为哮喘的防治提供一条新的途径。方法以酶消化法分离培养正常与哮喘豚鼠气道平滑肌细胞(ASMCs),应用RT-PCR方法测定每组细胞RyR各亚型RyR1,RyR2和RyR3的mRNA表达。结果正常与哮喘豚鼠ASMCs中,RyR1和RyR2的mRNA均见表达,未见RyR3mRNA的表达,并以表达RyR2mRNA为主。哮喘时,RyR1和RyR2的mRNA相对吸光度值分别为0.43±0.05和1.5±0.6,明显高于正常组RyR1mRNA的吸光度值0.34±0.03(n=6,P<0.01)。结论RyR1mRNA表达上调可能与哮喘发病相关。     

上皮间质转化与哮喘气道重塑的研究进展

支气管哮喘是一类以气道重塑为病理基础的呼吸道疾病,反复的炎性浸润与组织损伤修复可导致气道重塑,目前有关气道重塑形成的机制尚不全面。研究表明,上皮间质转化在气道重塑的发生和发展过程中发挥重要作用。气道上皮可通过分泌多种因子及信号通路诱发间质转化,进而导致哮喘气道重塑。该文对上皮间质转化与哮喘气道重塑之间的研究进行综述,为临床后续哮喘治疗和研究提供参考。     

气道上皮细胞铁死亡在哮喘中的作用:现状与展望

铁死亡是近年提出的以细胞内铁依赖的脂质过氧化为主要特征的细胞死亡方式,其机制主要涉及脂质过氧化、铁累积和抗氧化系统失衡。近年来,有关铁死亡与哮喘的研究正逐渐深入。阐明铁死亡调控哮喘的分子机制,有助于拓宽对哮喘病理机制的理解。本文从脂质过氧化、铁累积和抗氧化系统失衡这3个角度,阐述气道上皮细胞铁死亡在哮喘发生发展过程中的作用,有望为哮喘治疗寻求新的靶点和策略。     

小气道功能变化在咳嗽变异性哮喘诊断中的意义

目的通过对咳嗽变异性哮喘（Cough variant asthma,CVA）患者组（A组）与支气管哮喘患者组（B组）及健康对照组（C组）患者进行肺功能检测,探讨小气道功能变化对CVA的诊断价值。方法采用美国Senders厂生产的VIASYS综合肺功能仪,对A、B组患者及正常对照检测肺功能有关指标。结果经方差分析,3组间各项指标均有显著差异。经t检验进行两组间比较,发现B组FEF25%、FEF50%、FEF75%及FEF25%～75%与A及C组比较,有显著差异（P〈0.01）;A组FEF25%与C组比较,显著降低（P〈0.05）。结论 A组与正常对照组比较小气道功能变化（FEF25%）存在差异,A组存在小气道早期功能变化,小气道功能检测对CVA诊断有帮助。     

紧密连接蛋白occludin、ZO-1在溃疡性结肠炎中的表达及其临床意义

目的研究溃疡性结肠炎患者肠道紧密连接蛋白occludin、ZO-1的表达及其临床意义。方法收集2014年1月—2017年10月在中国医科大学附属盛京医院住院的溃疡性结肠炎患者临床资料和组织标本作为观察组,共80例,手术切除后经病理学证实的正常断端结肠组织20例,作为对照组。观察组按照Mayo临床评分(MCS)分为临床缓解组、轻度活动期组、中度活动期组、重度活动期组。免疫组织化学法检测肠道紧密连接蛋白occludin、ZO-1表达情况,并分析其与临床参数的相关性。结果 MCS和Mayo内镜评分(MCSe)在临床缓解组、轻度活动期组、中度活动期组、重度活动期组呈上升趋势,各组间差异均具有统计学意义(P0.05)。与对照组比较,各观察组均显著升高(P0.05),Geboes指数在各组中呈上升趋势,各组间差异均具有统计学意义(P0.05)。与对照组比较,紧密连接蛋白(occludin和ZO-1)在观察组中的表达下降。ZO-1、occludin的表达与MCS、MCSe、Geboes指数、C-反应蛋白(CRP)呈负相关。结论紧密连接蛋白ZO-1、occludin可能会成为评估溃疡性结肠炎患者肠道炎症程度和预测黏膜愈合的指标。     

热休克蛋白70在哮喘小鼠气道炎症模型中的表达及意义

目的 观察哮喘小鼠肺组织中热休克蛋白70(HSP70)的表达及其在哮喘中的作用.方法 将32只BALB/c小鼠随机分为N(正常对照组)、A(急性哮喘组I)、B(急性哮喘组II)和C(慢性哮喘组)4组,每组8只;卵蛋白致敏和激发建立哮喘小鼠模型;支气管肺泡灌洗液(BALF)分析和HE染色鉴定哮喘模型;免疫组化及Western blot法测定各组小鼠肺组织中HSP70的表达和定位.结果 哮喘组BALF嗜酸性粒细胞绝对计数及分类计数较N组皆有显著增高(P＜0.05),HE染色提示哮喘组与N组相比出现嗜酸性粒细胞浸润增多、纤毛脱失、平滑肌细胞层增厚等改变.HSP70在哮喘各组小鼠中主要定位在支气管上皮细胞、部分肺泡上皮细胞和炎症细胞中,而在N组中表达呈阴性结果;HSP70在哮喘组表达量较N组为高(P＜0.05),B组表达量显著高于A组(P＜0.05).结论 哮喘小鼠肺组织中HSP70主要表达于支气管上皮细胞、肺泡上皮细胞和炎症细胞中,参与哮喘发病.     

中药川贝对哮喘模型小鼠气道重塑及TGF-β1/Smad信号通路的影响

目的：观察中药川贝对哮喘模型小鼠气道重塑及转化生长因子-β1（TGF-β1）/Smad信号通路的影响,探讨其治疗哮喘的作用机制。方法：BALB/c小鼠,随机分为正常组、模型组、高剂量组、低剂量组、阳性对照组。除正常组外,其他各组用卵蛋白（OVA）建立小鼠哮喘模型。造模成功后高剂量组和低剂量组分别按18.0 mg·kg^-1和9.0 mg·kg^-1剂量给予中药川贝灌胃;阳性对照组雾化吸入0.5 mg·kg^-1地塞米松;正常组和模型组等量生理盐水灌胃,每天 1次,连续28 d。观察各组小鼠支气管管壁厚度和平滑肌厚度的变化以及支气管肺泡灌洗液（BALF）中嗜酸性粒细胞（EOS）计数的变化;酶联免疫吸附剂测定（ELISA）检测BALF和血清中TGF-β1浓度;蛋白印记分析（Western-blot）检测肺组织TGF-β1、磷酸化Smad2（p-Smad2）、Smad2、磷酸化Smad3（p-Smad3）、Smad3的表达;实时荧光定量PCR（Real Time-PCR）检测肺组织TGF-β1 mRNA、Smad2 mRNA、Smad3 mRNA的表达。结果：模型组小鼠支气管管壁厚度和平滑肌厚度,BALF中EOS计数,BALF和血清中TGF-β1浓度,肺组织TGF-β1、p-Smad2、Smad2、p-Smad3、Smad3的表达,肺组织TGF-β1 mRNA、Smad2 mRNA、Smad3 mRNA的表达均明显高于对照组（P<0.01）,高剂量组,低剂量组和阳性对照组上述指标则明显低于模型组（P<0.05或P<0.01）。结论：中药川贝可改善哮喘模型小鼠气道重塑状态,其机制可能与其抑制TGF-β1/Smad信号通路有关。     

Anthocyanins inhibit airway inflammation and hyperresponsiveness in a murine asthma model.

Asthma is a common chronic inflammatory disease regulated by coordination of T-helper cell type 2 (Th2) cytokines and inflammatory signal molecules. Additionally, oxidative stress may play an important role in airway inflammation such as eosinophilia, mucus hypersecretion, and airway hyperresponsiveness (AHR). In the present report, we investigated whether anthocyanins would reduce airway inflammation in a mouse asthma model immunized and challenged with ovalbumin (OVA). OVA inhalation elicited inflammatory responses characterized by eosinophilia and increased lipid hydroperoxide (LPO) in bronchoalveolar lavage (BAL) fluid, enhanced pause (Penh), increased glycoprotein and proliferating cell nuclear antigen (PCNA) expressions in mucus hypersecretion, and an increased expression of various cytokines and cyclooxygenase (COX) 2 in lung tissues. All parameters were attenuated in a dose-dependant manner by the administration of anthocyanins. These results suggest that anthocyanins may attenuate the development of asthma by downregulating Th2 cytokines, proinflammatory cytokines, and COX-2. Our findings suggest that anthocyanins have positive contributions as a dietary supplement for the prevention of asthma.     

Potential candidate genomic biomarkers of drug induced vascular injury in the rat

Drug-induced vascular injury is frequently observed in rats but the relevance and translation to humans present a hurdle for drug development. Numerous structurally diverse pharmacologic agents have been shown to induce mesenteric arterial medial necrosis in rats, but no consistent biomarkers have been identified. To address this need, a novel strategy was developed in rats to identify genes associated with the development of drug-induced mesenteric arterial medial necrosis. Separate groups (n = 6/group) of male rats were given 28 different toxicants (30 different treatments) for 1 or 4 days with each toxicant given at 3 different doses (low, mid and high) plus corresponding vehicle (912 total rats). Mesentery was collected, frozen and endothelial and vascular smooth muscle cells were microdissected from each artery. RNA was isolated, amplified and Affymetrix GeneChip® analysis was performed on selectively enriched samples and a novel panel of genes representing those which showed a dose responsive pattern for all treatments in which mesenteric arterial medial necrosis was histologically observed, was developed and verified in individual endothelial cell- and vascular smooth muscle cell-enriched samples. Data were confirmed in samples containing mesentery using quantitative real-time RT-PCR (TaqMan™) gene expression profiling. In addition, the performance of the panel was also confirmed using similarly collected samples obtained from a timecourse study in rats given a well established vascular toxicant (Fenoldopam). Although further validation is still required, a novel gene panel has been developed that represents a strategic opportunity that can potentially be used to help predict the occurrence of drug-induced mesenteric arterial medial necrosis in rats at an early stage in drug development.     

The Cdc42 inhibitor secramine B prevents cAMP-induced K+ conductance in intestinal epithelial cells

Cyclic AMP- (cAMP) and calcium-dependent agonists stimulate chloride secretion through the coordinated activation of distinct apical and basolateral membrane channels and ion transporters in mucosal epithelial cells. Defects in the regulation of Cl- transport across mucosal surfaces occur with cystic fibrosis and V. cholerae infection and can be life threatening. Here we report that secramine B, a small molecule that inhibits activation of the Rho GTPase Cdc42, reduced cAMP-stimulated chloride secretion in the human intestinal cell line T84. Secramine B interfered with a cAMP-gated and Ba2+-sensitive K+ channel, presumably KCNQ1/KCNE3. This channel is required to maintain the membrane potential that sustains chloride secretion. In contrast, secramine B did not affect the Ca2+-mediated chloride secretion pathway, which requires a separate K+ channel activity from that of cAMP. Pirl1, another small molecule structurally unrelated to secramine B that also inhibits Cdc42 activation in vitro, similarly inhibited cAMP-dependent but not Ca2+-dependent chloride secretion. These results suggest that Rho GTPases may be involved in the regulation of the chloride secretory response and identify secramine B an inhibitor of cAMP-dependent K+ conductance in intestinal epithelial cells.     

The regulatory and modulatory roles of TRP family channels in malignant tumors and relevant therapeutic strategies

Transient receptor potential (TRP) channels are one primary type of calcium (Ca²⁺) permeable channels, and those relevant transmembrane and intracellular TRP channels were previously thought to be mainly associated with the regulation of cardiovascular and neuronal systems. Nowadays, however, accumulating evidence shows that those TRP channels are also responsible for tumorigenesis and progression, inducing tumor invasion and metastasis. However, the overall underlying mechanisms and possible signaling transduction pathways that TRP channels in malignant tumors might still remain elusive. Therefore, in this review, we focus on the linkage between TRP channels and the significant characteristics of tumors such as multi-drug resistance (MDR), metastasis, apoptosis, proliferation, immune surveillance evasion, and the alterations of relevant tumor micro-environment. Moreover, we also have discussed the expression of relevant TRP channels in various forms of cancer and the relevant inhibitors’ efficacy. The chemo-sensitivity of the anti-cancer drugs of various acting mechanisms and the potential clinical applications are also presented. Furthermore, it would be enlightening to provide possible novel therapeutic approaches to counteract malignant tumors regarding the intervention of calcium channels of this type.