塔里木兔肾脏水通道蛋白在适应干旱环境中的作用

We measured the expression of aquaporin (AQP)1 -3 in kidneys of yarkand hares (Lepus yarcandensis )to understand the role of AQPs in adaptation to drought environment. We used H. E staining methods for detecting the histological structure of kidneys,and immunohistochemistry and western blotting for detecting expression of AQP1 - 3 in kidneys,
and also compared the results with those from domestic rabbits. Results showed that AQP1 is localized in capillary endothelial cell cytoplasm in glomeruli,and continued uninterrupted from proximal straight tubules into descending thin limbs in the outer medulla. AQP2 is observed in epithelial cells membrane in collecting ducts. AQP3 is localized in connecting tubules of the cortex and the outer medulla and epithelial cell basal membrane in collecting ducts. The expression of AQP1 -3 in kidneys is greater and the protein content is higher in yarkand hares than in domestic rabbits. These results indicate that the yarkand hare has increased the expression of aquaporins in kidneys,strengthened renal reabsorption of water and the ability for concentrating the urine,improving the ability for adaptation to arid environment.     

塔里木兔与家兔胰腺水通道蛋白表达的比较

通过检测塔里木兔（Lepus yarcandensis）胰腺中水通道蛋白（aquaporin,AQP）1和4的表达及分布情况,以探讨水通道蛋白在塔里木兔适应干旱缺水环境中的作用。采用常规 H.E染色观察塔里木兔胰腺组织学结构,采用免疫组织化学法检测AQP1和AQP4在胰腺中的分布位置及表达,并与家兔（Oryctolagus curiculus）进行比较。结果显示,AQP1在胰腺微血管内皮细胞、血细胞、泡心细胞和小叶内导管上皮细胞均有表达;AQP4在小叶间导管基底膜和胰岛细胞膜上有表达。与家兔相比,AQP1 在塔里木兔胰腺外分泌部的表达较弱,而在小叶内导管的表达较强;AQP4在塔里木兔胰腺内分泌部的表达较低。以上结果说明,AQP1在塔里木兔胰腺小叶内导管的表达上调,推测可能加强了浓缩胰液的能力,以尽量保住体内的水分,这是塔里木兔对干旱缺水环境的适应性调节。与家兔相比,塔里木兔胰腺AQP1和AQP4的表达均较低,说明塔里木兔胰腺水液代谢能力比家兔低,这可能与塔里木兔所食食物营养匮乏有关。     

塔里木兔胰腺水通道蛋白的表达和作用

通过检测塔里木兔(Lepus yarcandensis)胰腺中水通道蛋白（aquaporin,AQP）1和4的表达和分布情况,以探讨水通道蛋白在塔里木兔适应干旱缺水环境中的作用,采用常规 H.E.染色观察塔里木兔胰腺组织学结构,采用免疫组织化学检测AQP1和AQP4在胰腺中的分布位置及表达,并与家兔进行比较。结果显示,AQP1在微血管内皮细胞,血细胞,泡心细胞和小叶内导管上皮细胞均有表达;AQP4在小叶间导管基底膜和胰岛细胞膜上有表达。与家兔相比,AQP1 在塔里木兔胰腺外分泌部的表达较弱,而在小叶内导管的表达较强;AQP4在塔里木兔胰腺内分泌部的表达较低。以上结果说明,AQP1在塔里木兔胰腺小叶内导管的表达上调,推测可能加强了浓缩胰液的能力,以尽量保住体内的水分,是塔里木兔对干旱缺水环境的适应性调节。与家兔相比,塔里木兔胰腺AQP1和AQP4的表达均较低,说明塔里木兔胰腺水液代谢能力比家兔低,这可能与塔里木兔所食食物营养匮乏有关。     

水通道蛋白AQP1,3,4,5在双峰驼肺中的表达

目的研究水通道蛋白AQPs在双峰驼肺中的表达情况,探讨双峰驼适应极干旱荒漠环境的呼吸生理机制。方法运用常规形态学统计方法和石蜡切片HE染色法对双峰驼肺组织形态结构进行统计与分析,免疫组化方法对双峰驼肺中AQPs的表达进行定位分析。结果双峰驼气管长且弯曲,肺较致密且含水量较黄牛高。免疫组化检测显示,在双峰驼肺中有AQP1、AQP3、AQP4和AQP5四种AQPs表达。其中,AQP1主要表达于肺毛细血管网、淋巴管以及气管上皮细胞顶膜面;AQP3主要表达于气管上皮基底细胞质膜上;AQP4主要分布于整个气管上皮杯状细胞基底侧细胞膜和肺泡Ⅱ型上皮细胞;AQP5表达于气管粘膜下腺腺体细胞管腔面和肺泡Ⅰ型上皮细胞膜上。结论呼吸道和肺组织形态学特征表明双峰驼对干旱沙漠环境具有很好的适应性,AQPs在双峰驼肺中的强烈表达,与气道润化、气道水平衡、气道表面液体层、肺内液体转运和肺内水平衡等生理过程有关,为其适应极干旱荒漠环境提供了分子生物学依据。     

牦牛水通道蛋白AQP1和AQP3基因克隆及在不同组织中表达和定位

水通道蛋白(Aquaporin,AQP)广泛存在于生物体的各组织部位,影响着生物体水代谢的过程.为进一步研究水通道蛋白1(AQP1)和水通道蛋白3(AQP3)生物学功能,本文对牦牛(Bos grunniens)不同组织中 AQP1和AQP3基因的表达与定位进行了研究.采用PCR方法扩增牦牛AQP1和AQP3基因,对其序...     

急性冷暴露对大鼠肺组织中水通道蛋白AQP-1和AQP-5表达的影响

目的：探讨急性冷暴露后肺组织超微结构变化以及对水通道蛋白-1（AQP-1）和AQP-5表达的影响。方法：12只健康雄性Wistar大鼠随机分为室温（23℃±2℃）对照组和-25℃ 2 h冷暴露组（n=6）;记录冷暴露后大鼠直肠温度;透射电镜观察肺组织超微结构改变;RT-PCR法和Western blot法测定大鼠肺组织AQP-1和AQP-5基因和蛋白的表达水平。结果：急性冷暴露后大鼠的体心温度与对照组相比,明显降低（P<0.05）;肺组织超微结构亦发生改变,基底膜明显增厚,肺泡I上皮细胞（AT-I）核固缩,肺泡Ⅱ上皮细胞（AT-Ⅱ）胞浆空泡化增多;冷暴露后大鼠肺组织AQP-1的基因和蛋白表达未见明显变化,AQP-5的基因和蛋白表达均显著降低（P<0.05）。结论：急性冷暴露肺组织AQP-5基因和蛋白表达降低与寒冷暴露引发肺组织结构损伤可能存在一定因果关系。     

水通道蛋白

水通道蛋白 (aquaporin,AQP)是对水专一的通道蛋白 ,普遍存在于动、植物及微生物中。它所介导的自由水快速被动的跨生物膜转运 ,是水进出细胞的主要途径。1　水通道蛋白的发现长期以来 ,普遍认为细胞内外的水分子是以简单的跨膜扩散方式来透过脂双层膜。后来由于在生物物理学研究中发现红细胞及近端肾小管对渗透压改变引起的水的通透性很高 ,很难单纯以弥散来解释。因此 ,一些学者推测水的跨膜转运除了简单扩散外 ,还存在某种特殊的机制 ,并提出了水通道的概念。1988年 ,Agre等在鉴定人类 Rh血型抗原时 ,偶然在红细胞膜上发现了 1种新的 2…     

水通道蛋白4在脑缺血后脑水肿中的作用

脑缺血是由于动脉阻塞或灌注不足导致大脑局部血流减少无法满足代谢需求产生的功能障碍。脑水肿是脑组织间或细胞内液体过度积聚的病理现象,是脑缺血后较为严重的并发症,将会导致颅内压升高,脑组织受压而神经功能受损,甚至死亡。水通道蛋白(aquaporin)是一类分布在细胞膜上的蛋白质家族,目前已发现有13种亚型,主要调节细胞内外水平衡且参与细胞迁移和信号传导等多个生理病理过程。水通道蛋白4(aquaporin-4,AQP4)主要分布在中枢神经系统中星形胶质细胞的终足上,在细胞毒性水肿和血管源性水肿的形成和消除中起双重作用,与脑缺血后脑水肿有密切关系。机体通过转录过程及翻译后修饰等多个水平调节AQP4的表达协调其功能。本文回顾了目前AQP4在脑缺血后作用的最新进展,力图为治疗脑卒中后脑水肿提供新的研究方向。     

水通道蛋白在细胞中的调控

水通道蛋白是一个具有跨膜运输水分子功能的蛋白家族．其功能受到细胞精细调控,以维持细胞正常的生理状态,水通道蛋白异常将导致相关疾病的发生。重点介绍水通道蛋白在细胞中的调控机理。     

水通道

水通道邱俭,陈宜张（第二军医大学生理学教研室,上海２００４３３）关键词水通道,水通道蛋白进出细胞的水转运对细胞稳态是重要的。水通过脂质双层的运动很慢,这是一个单纯扩散过程,活化能高（＞１０ｋｃａｌ／ｍｏｌ）,且对汞化合物不敏感。过去曾认为其他一些细胞...     

塔里木兔和家兔的消化酶活性比较

为比较塔里木兔(Lepus yarcandensis)和家兔(Oryctolagus curiculus)消化能力的差异,探索塔里木兔对野外生存环境食物的适应机制,测定了塔里木兔和家兔胰腺及肠道的消化酶活性,包括淀粉酶(碘-淀粉比色法)、纤维素酶(3,5-二硝基水杨酸法)、脂肪酶(比浊法)及胰蛋白酶(紫外线吸收法)。用SPSS 15.0软件对实验数据进行统计学分析。采用one-way AVNOVA和多因素方差分析对比分析了塔里木兔和家兔消化酶活性。结果表明:1)塔里木兔肠道的淀粉酶活性明显高于家兔。其中,塔里木兔十二指肠、空肠及回肠的淀粉酶活性极显著高于家兔(P0.01)。淀粉酶活性在塔里木兔和家兔空肠最高。2)塔里木兔肠道的纤维素酶活性高于家兔。其中,塔里木兔盲肠的纤维素酶活性极显著高于家兔(P0.01),其回肠的纤维素酶活性显著高于家兔(P0.05)。纤维素酶活性在塔里木兔和家兔盲肠最高。3)家兔胰腺和肠道的脂肪酶活性明显高于塔里木兔。其中,家兔胰腺、十二指肠、空肠及回肠的脂肪酶活性显著高于塔里木兔(P0.05)。与消化道相比,脂肪酶活性在家兔和塔里木兔胰腺最高,肠道中空肠的脂肪酶活性最高。4)家兔胰腺和肠道的胰蛋白酶活性高于塔里木兔。其中,家兔空肠和十二指肠的胰蛋白酶活性显著高于塔里木兔(P0.05)。胰蛋白酶活性在家兔和塔里木兔空肠最高。可见,与家兔相比,塔里木兔对淀粉类、纤维类物质有较强的消化能力,对脂肪类的消化能力较弱。其中,淀粉酶活性和纤维素酶活性升高,脂肪酶活性和胰蛋白酶活性降低。这可能是塔里木兔适应贫瘠、匮乏的食物环境的重要原因之一。     

Cardiovascular Involvement in Connective Tissue Disease: The Role of Interstitial Lung Disease

Objective

The aim of this study was to assess cardiovascular involvement in patients with connective tissue disease (CTD), and determine whether interstitial lung disease (ILD) in these patients is associated with elevated cardiovascular risk.

Methods

This study evaluated a retrospective cohort of 436 CTD patients admitted to a large teaching hospital in Zhejiang province, China, along with an additional 436 participants of an annual community health screening conducted in the physical examination center who served as age- and gender-matched controls. Demographic, clinical, serologic and imaging characteristics, as well as medications used by each participant were recorded. Cardiovascular involvement was defined by uniform criteria. Correlations between clinical/serologic factors and cardiovascular involvement were determined by univariate and multivariate analyses.

Results

CTD patients had a significantly higher cardiovascular involvement rate than controls (64.7% vs 23.4%), with higher rates of diabetes, hypertension, and hyperlipidemia, elevated systolic and diastolic pressures, C-reactive protein, total cholesterol, and low-density lipoprotein cholesterol, and lower albumin and high-density lipoprotein cholesterol (all p < 0.05). Furthermore, CTP patients with cardiovascular involvement were significantly older, had higher systolic and diastolic pressures, C-reactive protein, glucose, and uric acid, higher rates of diabetes, hypertension, and use of moderate- to high-dose glucocorticoids, and longer disease duration compared to patients without involvement (all p < 0.05). Moreover, CTD in patients with cardiovascular involvement was more likely to be complicated by ILD (p < 0.01), which manifested as a higher alveolar inflammation score (p < 0.05). In the multivariate analysis, cardiovascular involvement in CTD patients was associated with age, systolic pressure, body mass index, uric acid, disease duration > 2 years, use of moderate- to high-dose glucocorticoids, and ILD with a high alveolar inflammation score.

Conclusion

Cardiovascular involvement is increased in CTD patients, and is associated with ILD with a higher alveolar inflammation score. Thus, early-stage echocardiography and CT scans should be used to detect potential cardiovascular complications in these patients.     

The Role of Aquaporins in pH-Dependent Germination of Rhizopus delemar Spores

Rhizopus delemar and associated species attack a wide range of fruit and vegetables after harvest. Host nutrients and acidic pH are required for optimal germination of R. delemar, and we studied how this process is triggered. Glucose induced spore swelling in an acidic environment, expressed by an up to 3-fold increase in spore diameter, whereas spore diameter was smaller in a neutral environment. When suspended in an acidic environment, the spores started to float, indicating a change in their density. Treatment of the spores with HgCl₂, an aquaporin blocker, prevented floating and inhibited spore swelling and germ-tube emergence, indicating the importance of water uptake at the early stages of germination. Two putative candidate aquaporin-encoding genes—RdAQP1 and RdAQP2—were identified in the R. delemar genome. Both presented the conserved NPA motif and six-transmembrane domain topology. Expressing RdAQP1 and RdAQP2 in Arabidopsis protoplasts increased the cells'' osmotic water permeability coefficient (P_f) compared to controls, indicating their role as water channels. A decrease in R. delemar aquaporin activity with increasing external pH suggested pH regulation of these proteins. Substitution of two histidine (His) residues, positioned on two loops facing the outer side of the cell, with alanine eliminated the pH sensing resulting in similar P_f values under acidic and basic conditions. Since hydration is critical for spore switching from the resting to activate state, we suggest that pH regulation of the aquaporins can regulate the initial phase of R. delemar spore germination, followed by germ-tube elongation and host-tissue infection.     

塔里木兔和家兔的消化酶活性比较研究

为比较塔里木兔(Lepus yarcandensis)和家兔（Oryctolagus curiculus）消化能力的差异,探索塔里木兔对野外生存环境的食物的适应机制,以家兔为对照,测定了塔里木兔和家兔的胰腺及肠道的消化酶—淀粉酶（碘-淀粉比色法）、纤维素酶（3,5-二硝基水杨酸法）、脂肪酶（比浊法）及胰蛋白酶（紫外线吸收法）的活性。用 SPSS 15.0 软件对实验数据进行统计学分析。结果表明：1）塔里木兔肠道的淀粉酶活性明显高于家兔。其中,塔里木兔十二指肠、空肠及回肠的淀粉酶活性极显著高于家兔（P ＜ 0.01）。淀粉酶活性在塔里木兔和家兔空肠最高。2）塔里木兔肠道的纤维素酶活性高于家兔。其中,塔里木兔盲肠的纤维素酶活性极显著高于家兔（P ＜ 0.01）,其回肠的纤维素酶活性显著高于家兔（P ＜ 0.05）。纤维素酶活性在塔里木兔和家兔盲肠最高。3）家兔胰腺和肠道的脂肪酶活性明显高于塔里木兔。其中,家兔胰腺、十二指肠、空肠及回肠的脂肪酶活性显著高于塔里木兔（P ＜ 0.05）。与消化道相比,脂肪酶活性在家兔和塔里木兔胰腺最高,肠道中空肠的脂肪酶活性最高。4）家兔胰腺和肠道的胰蛋白酶活性高于塔里木兔。其中,家兔空肠和十二指肠的胰蛋白酶活性显著高于塔里木兔（P ＜ 0.05）。胰蛋白酶活性在家兔和塔里木兔空肠最高。可见,与家兔相比,塔里木兔对淀粉类、纤维类物质有较强的消化能力,对脂肪类的消化能力较弱。其中,淀粉酶活性和纤维素酶活性升高,脂肪酶活性和胰蛋白酶活性降低。这可能是塔里木兔适应贫瘠、匮乏的食物环境的重要原因之一。     

Genome-Wide Identification and Expression Analysis of Aquaporins in Tomato

The family of aquaporins, also called water channels or major intrinsic proteins, is characterized by six transmembrane domains that together facilitate the transport of water and a variety of low molecular weight solutes. They are found in all domains of life, but show their highest diversity in plants. Numerous studies identified aquaporins as important targets for improving plant performance under drought stress. The phylogeny of aquaporins is well established based on model species like Arabidopsis thaliana, which can be used as a template to investigate aquaporins in other species. In this study we comprehensively identified aquaporin encoding genes in tomato (Solanum lycopersicum), which is an important vegetable crop and also serves as a model for fleshy fruit development. We found 47 aquaporin genes in the tomato genome and analyzed their structural features. Based on a phylogenetic analysis of the deduced amino acid sequences the aquaporin genes were assigned to five subfamilies (PIPs, TIPs, NIPs, SIPs and XIPs) and their substrate specificity was assessed on the basis of key amino acid residues. As ESTs were available for 32 genes, expression of these genes was analyzed in 13 different tissues and developmental stages of tomato. We detected tissue-specific and development-specific expression of tomato aquaporin genes, which is a first step towards revealing the contribution of aquaporins to water and solute transport in leaves and during fruit development.     

NDRG2在大鼠肺缺血再灌注损伤中的表达

目的:通过建立大鼠肺缺血再灌注损伤(Lung ischemia-reperfusion injury,LIRI)模型,观察肺缺血再灌注损伤后,肺组织中N-myc下游调节基因2(N-myc downstream regulated gene,NDRG2)表达水平的变化.方法:将70只健康成年雄性SD大鼠随机分成对照组(C)、缺血组(I)、缺血再灌注组(I/R)(后两组各含3个亚组),每组10只.麻醉固定大鼠,颈部切口行气管插管.右侧开胸,肺缺血组依次分别选择游离夹闭右肺门(即右主支气管,右肺动、静脉)缺血30 min、60 min、120 min后,麻醉处死大鼠获取肺组织.肺缺血再灌注组同样选择游离夹闭右肺门,于夹闭右肺门60 min后松开,分别取再灌注30 min、60 min、120m in后麻醉处死大鼠获取肺组织样本.采用免疫组化对肺组织NDRG2进行蛋白定位检测、RT-PCR对肺组织NDRG2 mRNA含量进行检测、Western-blot对肺组织NDRG2蛋白含量进行检测.结果:肺缺血组与对照组比较,肺组织NDRG2的表达无明显变化(P＞0.05);肺缺血再灌注组与对照组比较,NDRG2蛋白含量和mRNA表达量逐渐下降,在60 min时达最低,之后又有所回升,但仍低于对照组(P＜0.05).结论:肺缺血再灌注损伤可下调肺组织中NDR G2的表达含量,NDRG2可能是肺缺血再灌注损伤的靶向调控位点.     

中药止泻液对乳鼠轮状病毒肠炎水通道蛋白表达的影响

目的:用中药止泻液对轮状病毒肠炎乳鼠进行干预后,检测乳鼠空肠,回肠与近端结肠粘膜水通道蛋白(aquaporin,AQP)1,3,4,8mRNA的表达情况,研究中药止泻液对乳鼠轮状病毒肠炎AQP1,3,4,8mRNA表达的影响.方法:在实验条件下,用恒河猴SAll株轮状病毒感染6日龄乳鼠24h后,予各乳鼠灌注止泻液,每日3次,于接毒后48 h始,每天处死2只乳鼠,连续6天.采用实时荧光定量PCR方法对乳鼠的空肠,回肠与近端结肠黏膜细胞的AQP1,3,4,8 mRNA进行定性和相对定量分析研究,探讨中药止泻液对轮状病毒腹泻模型乳鼠肠道AQP1,3,4,8mRNA表达的影响.结果:结肠AQP1 mRNA在中药干预组乳鼠中表达量显著高于空白组(t=-3.47,P<0.05). AQP3,4,8mRNA表达量无统计学差异.在空肠,回肠中,中药干预组与空白组AQP1,3,4,8 mRNA袁达量均无统计学差异.结论:中药干预组乳鼠结肠AQP1 mRNA表达上升,提示中药止泻液能明显提高乳鼠结肠组织AQP1 mRNA的表达,通过增加结肠对水分的吸收,来达到收敛止泻的作用.在本研究中,中药止泻液对轮状病毒肠炎乳鼠肠道AQP3,4,8 mRNA表达没有明显的影响.     

波形蛋白在非小细胞肺癌组织中的表达及其临床意义

目的:探究波形蛋白在非小细胞肺癌(NSCLC)组织中的表达及其与肺癌浸润转移的相关性。方法:收集2012年6月-2014年6月我院手术切除的NSCLC癌组织标本150例及癌旁正常组织(距肿瘤5 cm)79例,提取两组的RNA,采用实时荧光定量聚合酶链反应(RT-PCR)检测波形蛋白m RNA表达水平,免疫组化法检测波形蛋白的蛋白表达,分析波形蛋白表达水平与淋巴结转移、TNM分期的相关性。结果:波形蛋白m RNA在NSCLC癌组织中的表达明显高于癌旁正常组织(P0.05)。NSCLC癌组织中波形蛋白m RNA表达水平的上调与淋巴结转移及TNM分期(P0.05)相关。结论:波形蛋白在NSCLC患者中表达异常升高,与NSCLC的发生和浸润转移密切相关。