低温诱发肉鸡肺血管重塑过程中肺动脉平滑肌细胞c-Myc蛋白表达的变化

试验旨在研究环境低温诱发肉鸡肺血管重塑过程中肺小动脉血管平滑肌细胞c-Myc蛋白的表达变化,初步探讨肉鸡肺血管重塑的发生机制.120只雄性AA商品代肉鸡15日龄时随机分为对照组((22±1.5)℃)条件下饲养)和低温组((11±2)℃条件下饲养).15～50日龄,每周每组随机取6只,取肺组织做石蜡切片,Weigert-间苯二酚复红染色,观察并测定m管重塑情况;采用免疫组织化学方法榆测肺动脉血管平滑肌细胞c-Myc蛋白表达,并对其进行半定量化.结果显示:(1)低温组肉鸡肺小动脉结构从36日龄开始较对照组发生了明显的重塑(P<0.01或P<0.05);(2)低温组肉鸡肺小动脉血管平滑肌细胞c-Myc蛋白表达从29日龄开始较对照组明显增加(P<0.01).结果表明:低温叫显诱发了肉鸡肺小动脉平滑肌细胞c-Myc蛋白的表达,且参与了肉鸡肺血管重塑的发生发展.     

c-Jun在肉鸡腹水综合征过程中肺动脉壁的表达变化及维拉帕米的干预研究

本试验研究低温诱发肉鸡腹水综合征过程中肺动脉壁c-Jun的表达,及阻断钙信号对肺动脉壁c-Jun表达的影响和对肉鸡腹水综合征发生的干预情况,为肉鸡腹水综合征发生机制的研究提供基础。240只15日龄AA肉鸡分为对照组、低温组和维拉帕米组,15~50日龄每组随机取6只测定右心肥大指数,取肺组织做石蜡组织切片进行c-Jun免疫组化染色分析。结果显示,29~50日龄低温组肉鸡右心肥大指数较对照组和维拉帕米组明显升高(P0.05),低温组肉鸡20~50μm、50~100μm和100~200μm肺小动脉c-Jun阳性指数分别从22、29日龄和29日龄开始到50日龄极显著高于对照组和维拉帕米组(P0.01)。研究结果认为,低温诱发肉鸡腹水综合征过程中不同直径大小肺小动脉壁cJun表达明显增强,阻断钙信号可明显抑制肺小动脉壁c-Jun的表达和肉鸡腹水综合征的发生。     

高盐促进肉鸡肺小动脉壁癌基因c-jun mRNA的表达

本试验旨在研究高盐诱发肉鸡腹水综合征（AS）过程中肺小动脉壁癌基因c-jun mRNA表达的规律，初步探讨肺小动脉壁癌基因c-jun mRNA表达在AS发生过程中的可能作用。将100只1日龄商品代AA雄性肉仔鸡常规育雏，8日龄起随机分为对照组和高盐组（饮水中添加0．30％氯化钠）。分别在15、22、29、36、43、50日龄时每组随机取6只鸡的肺组织制备切片，以原位杂交法结合图像分析法测定肺小动脉壁癌基因c-jun mRNA的表达。结果表明：对照组肉鸡生长发育过程中肺小动脉壁癌基因c-jun mRNA表达呈现动态变化，22日龄是其表达高峰。15、29、36日龄时高盐组各级肺小动脉壁c-jun mRNA相对含量极显著高于对照组（P〈0．01）。43、50日龄时高盐组仅外径20～50μm肺小动脉壁c-jun mRNA相对含量极显著高于对照组（P〈0．01）。提示高盐促进了肉鸡肺小动脉壁癌基因c-jun mRNA表达，尤其是在外径20～50μm的肺小动脉壁。肺小动脉壁癌基因c-jun mRNA的过度表达可能参与了高盐诱发AS的过程。     

低温诱发肉鸡腹水综合征发生过程中肺血管重塑与肺动脉高压的关系

将160只15日龄雄性AA商品代肉鸡随机分为对照组((22±1.5)℃)和低温组((11±2)℃)。15~50日龄,每组每周随机取6只,以右心导管法直接测定肺动脉压(PAP),并取肺组织做石蜡切片进行Weigert-间苯二酚复红染色,分析肺血管重塑情况。50日龄时统计整个饲养过程中肉鸡腹水综合征(AS)发生率。结果显示,低温组肉鸡AS发生率(18.75%)极显著高于对照组(1.25%)(P<0.01);低温组肉鸡PAP从22日龄开始较对照组明显升高(P<0.01或P<0.05);20~50、50~100、100~200μm肺动脉结构分别从36、43、43日龄开始较对照组发生了明显的重塑(P<0.01或P<0.05)。结果表明,低温诱发肉鸡AS过程中出现了明显的PAP升高和肺血管重塑,且肺动脉高压(PH)促进了肺血管重塑,肺血管重塑加剧了PAP升高。     

低温诱发肉鸡腹水综合征过程中肺动脉平滑肌细胞增殖的变化

本试验旨在研究低温诱发肉鸡腹水综合征(AS)过程中平滑肌细胞增殖在肺血管重塑过程中的动态变化,初步探讨肺血管重塑的机制。160只15日龄雄性AA商品代肉鸡随机分为对照组(22℃±1.5℃)和低温组(11℃±2℃)。试验持续至50日龄,期间每周每组随机取6只,取肺组织做石蜡切片,Weigert-间苯二酚复红染色,观察并测定血管重塑情况;采用免疫组织化学方法检测肺动脉增殖细胞核抗原(PCNA)蛋白表达,并对其半定量化,分析平滑肌细胞增殖情况。50日龄时统计整个饲养过程中AS发生率。结果显示:(1)低温组肉鸡AS发生率(18.75%)极显著高于对照组(1.25%)(P<0.01);(2)低温组肉鸡直径20-50μm和50-150μm肺动脉结构从36日龄开始较对照组发生了明显的重塑(P<0.01或P<0.05);(3)低温组肉鸡直径20-50μm和50-150μm的肺动脉平滑肌细胞增殖指数分别从22日龄和36日龄开始极显著高于对照组(P<0.01)。结果表明:低温诱发肉鸡AS过程中肺动脉结构发生了明显的重塑,肺动脉平滑肌细胞发生了明显的增殖,并且平滑肌细胞增殖促进了肺血管重塑,在肉鸡AS发生发展过程中起着重要的作用。     

红三叶异黄酮对肉鸡肺血管重构和平滑肌细胞凋亡的影响

为观察在低温环境下红三叶异黄酮对肉鸡肺血管重构和平滑肌细胞凋亡的影响,选取270只10日龄肉鸡,随机分为常温对照组(Ⅰ)、低温组(Ⅱ)和低温黄酮组(Ⅲ),每组6个重复,每个重复15只,Ⅰ组和Ⅱ组饲喂基础饲粮,Ⅲ组饲喂添加红三叶异黄酮20mg/kg的试验饲粮,分别于14,21,28,35,42日龄从各组随机抽取6只肉鸡,观察各组肉鸡肺血管病理形态学变化,检测Caspase-3和Bcl-2蛋白表达情况。结果表明,Ⅱ组肉鸡肺小动脉结构在28,35,42日龄比Ⅰ组肺小动脉管壁有不同程度增厚、管腔变窄,发生了明显的重构现象(P0.05或0.01)。Ⅲ组与Ⅱ组相比,在28,35,42日龄肺血管重构现象减轻(P0.05或0.01),肉鸡肺动脉高压综合征(PHS)发病率降低,血管平滑肌细胞Caspase-3蛋白阳性率增高,Bcl-2蛋白表达下降;表明红三叶异黄酮能有效减轻肉鸡肺血管重构,其机制可能与调节平滑肌细胞凋亡有关。     

BQ123对低温诱发的肉鸡肺血管重塑的影响

动态观察了肺小动脉中膜平滑肌的增殖及管腔面积的改变,探讨内皮素受体拮抗剂BQ123 对低温诱发的肉鸡肺血管重塑的影响。200只16日龄AA肉鸡随机均分为4组: 常温(20 ℃)对照组、低温(7~9 ℃)组、低温低剂量BQ123组和低温高剂量BQ123组。23日龄、30日龄时测定肺动脉压,并取肺组织做石蜡切片, 以Weigert间苯二酚复红染色,形态学计算机图象分析法测定肺细小动脉外径和内径、管总面积和管腔面积,计算中膜厚度占外径百分值(%)mMTPA及管壁面积/管总面积WA/TA(%)。结果显示:(1)BQ123 抑制低温肉鸡平均肺动脉压的升高,23日龄时低温高剂量BQ123组显著低于低温组(P<0 05),30 日龄时低温低剂量BQ123 组和低温高剂量BQ123组均极显著低于低温组(P<0 01);(2)BQ123 抑制低温肉鸡肺小动脉WA/TA(%)的升高,30~50μm的肺小动脉,低温组极显著高于低温低剂量BQ123 组及低温高剂量BQ123 组(P<0 01),其它分级的肺小动脉组间差异性与此类似;(3) BQ123抑制低温肉鸡肺小动脉的mMTPA的升高,30~50μm的肺小动脉,23日龄时低温组极显著高于低温低剂量BQ123 组和低温高剂量BQ123 组(P< 0 01), 30 日龄时低温组显著高于低温低剂量BQ123组(P<0 05)、极显著高于低温高剂量BQ123组(P<0 01),30μm以下的肺小动脉组间差异性与此类似,50~120μm     

BQ123抑制低温诱发肺动脉高压肉鸡的肺小动脉肌型化

本试验用高选择性内皮素A受体拮抗剂BQ123处理低温肉鸡的方法,通过动态观察肺动脉压及肌型、部分肌型及非肌型肺小动脉数量的变化,探讨内源性内皮素在肺血管重塑中的作用。结果显示:30日龄时,低温组平均肺动脉压极显著高于常温组、低BQ123组及高BQ123组(P<0.01);低温组肌型及部分肌型肺小动脉的占位比均极显著高于常温组、低BQ123组及高BQ123组(P<0.01);低温组非肌型肺小动脉占位比极显著低于常温组、低BQ123组和高BQ123组(P<0.01),高BQ123组显著高于低BQ123组(P<0.05)。结果表明,BQ123可明显抑制低温肉鸡肺动脉高压和非肌型肺小动脉的肌型化的发生发展,因此认为内源性ET在低温诱发肺动脉高压肉鸡肺血管重塑的发生发展过程中起重要作用。     

缺氧与肉鸡肺动脉平滑肌细胞c-fos和 c-myc基因表达的关系

利用细胞培养、原位杂交、免疫组化、图像分析等技术研究缺氧与肉鸡肺动脉平滑肌细胞癌基因c-fos和c-myc表达的关系,进一步阐明缺氧与以肺动脉增殖重塑为特征的肉鸡腹水综合征的关系。结果显示,缺氧显著引起肺动脉平滑肌细胞内癌基因c-fos和c-mycmRNA的表达(c-fosmRNA:常氧组为144.6±20.2,缺氧组为198.1±32.8,P<0.01;c-mycmRNA:常氧组为125.4±18.8,缺氧组为167.1±22.4,P<0.01)。缺氧显著引起肺动脉平滑肌细胞内癌基因c-fos和c-myc蛋白的表达(c-fos蛋白:常氧组为150.9±33.2,缺氧组为225.9±37.0,P<0.01;c-myc蛋白:常氧组为162.1±28.5,缺氧组为228.8±33.4,P<0.01)。结果表明缺氧能够明显诱发肉鸡肺动脉平滑肌细胞内癌基因c-fos、c-myc的转录和表达,是启动肺动脉平滑肌细胞增殖的重要原因。本研究阐明缺氧是以肺动脉重塑为特征的肉鸡腹水综合征的主要诱因。     

维拉帕米对肉鸡腹水综合征患鸡心肌细胞c-fos和c-jun表达的影响

右心肥大衰竭是导致腹水综合征(ascites syndrome,AS)患鸡发病的重要因素.作者通过免疫组化和原位杂交技术初步探讨c-fos和c-jun在AS肉鸡右心肥大和衰竭过程中的变化及维拉帕米对其表达的影响.结果发现:在22、29、36、43和50日龄时,与对照组肉鸡相比,低温组肉鸡右心室心肌细胞c-fos/c-jun蛋白和mRNA的平均光密度值显著或极显著(P<0.05;P<0.01)升高,其mRNA的阳性面积百分比也极显著升高(P<0.01),与低温组肉鸡相比,维拉帕米组肉鸡右心室心肌细胞c-fos/c-jun蛋白和mRNA的平均光密度值显著或极显著(P<0.05;P<0.01)降低,其mRNA的阳性面积百分比也极显著降低(P<0.01).结果表明c-fos和c-jun癌基因在右心肥大和衰竭中起着重要作用.维拉帕米可有效抑制c-os和c-jun癌基因的表达,有利于防止右心肥大和功能性衰竭.     

Characteristics of the in vitro hypoxic pulmonary vasoconstrictor response in isolated equine and bovine pulmonary arterial rings

OBJECTIVE: Hypoxaemia accompanies dorsal recumbency in the horse and frequently complicates general anaesthesia. The physiology associated with this phenomenon is poorly understood. One possible cause of poor tolerance to dorsal recumbency is an absent or reduced response to hypoxic pulmonary vasoconstriction (HPV). This study compared the HPV response in isolated pulmonary artery vessels from equivalent regions of equine and bovine lung. ANIMALS: Equine and bovine, in vitro study. MATERIALS AND METHODS: Equine and bovine pulmonary arteries were removed from the lungs of euthanased horses and cattle. Measurements of isometric tension were made on isolated rings of pulmonary vessels at 37 degrees C in a Krebs' saline solution. Hypoxia was induced by bubbling with a nominally 0% O(2) gas mixture. RESULTS: A significant HPV response was observed above a baseline tension induced by phenylephrine (PE; 0.3 microm) or 5-hydroxytryptamine (5-HT; 0.1 microm). The HPV response in equine pulmonary vessels was approximately 33% less than the response observed in equivalent bovine vessels (equine 196 +/- 20%versus bovine 290 +/- 32%; p < 0.05). Removal of the endothelium (by rubbing the luminal surface) significantly reduced but did not abolish the HPV response. Incubation with the nitric oxide (NO) synthase inhibitor N-nitro-l-arginine methyl ester (L-NAME; 100 microm), or COX-1/COX-2 inhibitor indomethacin (10 microm) markedly attenuated the HPV response in equine vessels. CONCLUSIONS: These results suggest that a significant HPV response exists in isolated equine pulmonary vessels; a component of this response requires a functional endothelium. Inhibition of cyclooxygenase and NO synthase attenuated the response, suggesting the involvement of a COX product and/or NO in mediating this effect either directly or indirectly. Alternatively, a non-COX related action of the nonsteroidal anti-inflammatory drug, indomethacin, may be involved.     

Effect of frusemide on transvascular fluid fluxes across the lung in exercising horses

Reasons for performing study: Frusemide (Fru) is widely prescribed for management of racehorses experiencing EIPH. The effect of Fru in the lung appears to be a reduction in transcapillary pressures and inhibition of the erythrocyte anion exchange, which may lead to attenuation of transpulmonary fluid fluxes during exercise. Hypothesis: Treatment with Fru will attenuate transpulmonary fluid fluxes in horses during high intensity exercise. Methods: In a crossover study, 6 race‐fit Standardbred horses were treated with 250 mg of Fru i.v. (FruTr) or placebo (Con) 4 h before exercise on a high speed treadmill until fatigue. Arterial and central mixed venous blood, as well as CO₂ elimination and O₂ uptake, were sampled. Volume changes across the lung and transvascular fluid fluxes were calculated from changes in haemoglobin, packed cell volume, plasma protein and cardiac output (Q). Results: During exercise, Q increased in both Con and FruTr, with Q being significantly lower in FruTr (mean ± s.e. 301.8 ± 8.5 l/min at fatigue) compared to Con (336.5 ± 15.6 l/min) (P<0.01). At rest frusemide had no effect on erythrocyte (J_ER) and transvascular (J_V‐A) fluid fluxes across the lung. Exercise had a significant effect on J_ER and J_V‐A (P≤0.02). During exercise, J_ER (at fatigue 14.6 ± 2.3 l/min and 11.6 ± 2.2 l/min in Con and FruTr, respectively) and J_V‐A (at fatigue14.9 ± 2.3 l/min and 12.0 ± 2.2 l/min in Con and FruTr, respectively) were not significantly different between Con and FruTr (P = 0.6 and P = 0.8 for J_ER and J_V‐A, respectively). Conclusions and clinical importance: Fru does not have a measurable effect on J_ER and J_V‐A. Cardiac output was reduced in FruTr, suggesting that there were also smaller changes in the capillary recruitment and transvascular transmural hydrostatic pressures; however, this did not effect J_V‐A. Therefore, Fru at the dose of 250 mg does not appear to be an effective treatment for regulating pulmonary transvascular forces during exercise in horses.     

Ventilation distribution assessed with electrical impedance tomography and the influence of tidal volume,recruitment and positive end-expiratory pressure in isoflurane-anesthetized dogs

Objective

To examine the intrapulmonary gas distribution of low and high tidal volumes (V_T) and to investigate whether this is altered by an alveolar recruitment maneuver (ARM) and 5 cmH₂O positive end-expiratory pressure (PEEP) during anesthesia.

扎鲁司特对低温诱发的肉鸡肺动脉高压的影响

将60只雄性艾维茵商品代肉鸡常规育雏，17日龄随机分为3组：常温对照组(常温组)、低温模型组(低温组)和低温扎鲁司特组(扎鲁司特组)。各组于处理后14d(31日龄)和21d(38日龄)分别用右心导管法及直接导管法测定扎鲁司特对肺动脉压(PAP)和股动脉压(FAP)的影响，同时测定红细胞压积(PCV)和腹水心脏指数(AHI)。结果：(1)在低温环境的早期，扎鲁司特使肉鸡肺动脉收缩压和舒张压均升高，后期仅使舒张压升高；(2)扎鲁司特使PCV升高，增加了血液黏性阻力；(3)扎鲁司特使AHI升高，促进右心肥大；(4)扎鲁司特对股动脉压无显著性影响。可见扎鲁司特促进了肉鸡肺动脉高压综合征的发生发展。     

Growth-related changes in the pulmonary function of goats

Growth-related changes in pulmonary function values were investigated in 20 healthy French Alpine goats, aged between 20 and 550 days, weighing 7–55 kg. Pulmonary ventilation, mechanics of breathing and arterial oxyten tension were measured using standardized techniques and methods adapted for goats of different body sizes. The Ppl values and the tI/tTOT ratio showed no significant changes with age and body size. The ventilation values (Vt, Ve, mVI and mVE) increased linearly with growth. There was a significant correlation of age and body weight with dynamic lung compliance (Cdyn), total pulmonary resistance (RL), viscous work of breathing (Wvis tot) and minute viscous work (Wvis min) throughout the age range studied. Cdyn, Wvis tot and Wvis min increased and RL decreased with age and body weight. Arterial blood gases (PaO₂ and PaCO₂) did not show significant changes over the age range studied. Regression equations for each pulmonary function parameter are given with body weight as the independent variable. Data for the mechanics of breathing were compared with those elsewhere for cattle, horses, man and dogs.     

L-精氨酸对肺动脉高压肉鸡肺小动脉平滑肌细胞凋亡的影响

将240羽14日龄艾维茵-2000商品代肉鸡于14d时随机等分为常温对照组（NT组），低温组（LT组）和低温加L-精氨酸组（LA），白14d起，LT和LA组舍内温度从28℃以1～2℃／d的速度下降，至21d降至12～14℃，并维持到试验结束。此外，LA组自14d起在饲料中按100mg／kg剂量添加L—Arg，直至试验结束。记录肺动脉高压综合征（PHS）发病率，并分别于24、32、39和45d测定右心室／全心室质量比（RV／TV）、血浆NO、肺小动脉管壁面积／管总面积（WA／TA）和平均中膜厚度（mMTPA），并用原位缺口末端标记法（TUNEL法）对肺小动脉凋亡平滑肌细胞进行染色和计数。结果：LT组肉鸡PHS发病率、RV／TV、mMTPA和WA／TA值较NT组升高（P〈0．05）；LA组肉鸡PHS发病率较LT组降低（P〈0．05），mMTPA和WA／TA值与LT组相比呈下降趋势，在32d时显著降低（P〈0．05）；LT组血浆N0浓度总体上与NT组差异不显著（P〉0．05），仅在39d时降低（P〈0．05），而LA组血浆N0浓度自32d时起较NT和LT组显著升高（P〈0．05）；LA组肉鸡肺小动脉平滑肌凋亡细胞百分率较NT组和LT组均显著升高（P〈0.05），但LT组与NT组之间相比无显著差异（P〉0．05）。上述结果表明，L-Arg／NO通过促进肺小动脉平滑肌细胞凋亡，从而在一定程度上抑制了肺血管重构的形成。     

一氧化氮对肉鸡肺血管重塑影响的研究进展

肺血管重塑是肉鸡肺动脉高压综合征的重要病理学变化特征,虽然对其形成的机制尚未完全清楚,但众多细胞因子在其形成过程中起重要作用已得到共识。一氧化氮作为一种信号物质,在血管扩张、细胞增殖与凋亡、内皮素分泌与合成以及抗氧化等过程中发挥着重要的调控作用,显示了其在抑制肉鸡肺血管重塑、预防和治疗肺动脉高压发生方面有着广阔的应用前景。