低温诱发肉鸡肺动脉高压中肺血管重塑的作用

采用低温处理肉仔鸡的方法,通过测定肺动脉压、肺小动脉中膜厚度占外径百分比及中膜面积与血管总面积比的变化,探讨肉鸡肺动脉高压的发生机制.结果表明:肺血管重塑在低温诱发肉鸡的肺动脉高压发生发展中起重要作用.     

l-Arginine inhibiting pulmonary vascular remodelling is associated with promotion of apoptosis in pulmonary arterioles smooth muscle cells in broilers

OBJECTIVE: Pulmonary vascular remodelling is one of the important pathological bases of broiler pulmonary hypertension syndrome (PHS). Nitric oxide (NO) has been found to inhibit proliferation and to induce apoptosis in pulmonary artery smooth muscle cells (SMC) in mammals with pulmonary hypertension. The present study was conducted to evaluate the effects of NO precursor l-arginine on pulmonary vascular remodelling in broilers with pulmonary hypertension induced by cold exposure and to examine whether NO-induced apoptosis in pulmonary arteriole SMC is involved in the regulatory mechanisms. METHODS: Two hundred and forty mixed-sex commercial broilers were equally assigned to three groups and reared in normal brooding temperatures before day 14. Starting on day 14 continuing until the end of the experiment, the control group was brooded in normal temperatures whereas the other two groups were subjected to low ambient temperatures with or without l-arginine added to the basal diets. Cumulative PHS mortality and body weight were recorded in each group. Right/total ventricle ratio (RV/TV), plasma NO concentration and pulmonary vascular morphological changes were analyzed. TdT-mediated dUTP-biotin nick-end labeling (TUNEL) assay was used to detect apoptosis in pulmonary arteriole SMC. RESULT: l-Arginine, in group A, had no effect on body weights under cold temperature condition. Birds kept in group B had increased PHS mortality, RV/TV ratio, vessel wall area/vessel total area ratios (WA/TA) and mean media thickness in pulmonary arterioles (mMTPA) (P<0.05). Percentages of apoptotic SMC in pulmonary arterioles in group B were not altered by cold exposure (P>0.05). Supplemental dietary l-arginine in group A elevated plasma NO level (P<0.05), reduced PHS mortality (P<0.05), attenuated pulmonary vascular remodelling and increased the percentages of apoptotic SMC (P<0.05) when compared with the group B. CONCLUSION: Supplemental l-arginine partially inhibited pulmonary vascular remodelling that occurred secondary to increased pulmonary pressure; NO-induced apoptosis in arteriole SMC might contribute to its regulatory effect on pulmonary vascular structural changes.     

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。     

Clinical and pathological characterisation of primary pulmonary hypertension in a dog

Primary pulmonary hypertension was diagnosed in an eight-year-old labrador retriever on the basis of echocardiographic findings of severe right ventricular eccentric hypertrophy, abnormally high systolic and diastolic pulmonary arterial pressures calculated by applying the modified Bernoulli equation to the tricuspid and pulmonary insufficiency peak velocities, and the absence of any underlying disease known to cause secondary pulmonary hypertension. The clinical abnormalities developed gradually, from exercise intolerance starting early in life to terminal right-sided congestive heart failure. Consistent histopathological findings were severe intimal and medial thickening of small arteries and arterioles that led to vascular obliteration.     

Calcium antagonists,diltiazem and nifedipine,protect broilers against low temperature‐induced pulmonary hypertension and pulmonary vascular remodeling

This study was designed to determine whether calcium antagonists, diltiazem and nifedipine, can depress low temperature‐induced pulmonary hypertension (PH) in broilers (also known as ascites) and to characterize their efficacy on hemodynamics and pulmonary artery function. Chicks were randomly allocated into six experimental groups and orally administered with vehicle, 5.0 mg/kg body weight (BW)/12 h nifedipine or 15.0 mg/kg BW/12 h diltiazem from 16 to 43 days of age under low temperature. The mean pulmonary arterial pressure (mPAP), the ascites heart index (AHI), the erythrocyte packed cell volume (PCV) and the relative percentage of medial pulmonary artery thickness were examined on days 29, 36 and 43. The data showed that administration of diltiazem protected broilers from low temperature‐induced pulmonary hypertension and vascular remodeling. Although nifedipine prevented mPAP from increasing during the early stage, it did not suppress the development of PH during the late stage and did not keep heart rate (HR), PCV, AHI and the thickness of pulmonary small artery smooth muscle layer at the normal levels. Taken together, our results showed that diltiazem can effectively prevent low temperature‐induced pulmonary hypertension in broilers with fewer side‐effects and may be a potential compound for the prevention of this disease in poultry industry.     

Bromodeoxyuridine labeling and DNA content of pulmonary arterial medial cells from hypoxia-exposed and nonexposed healthy calves.

Vascular medial thickening is a prominent finding in people and animals with refractory neonatal pulmonary hypertension. Smooth muscle cells are capable of 2 distinct growth responses in vivo: hypertrophy or hyperplasia. Hypertrophic smooth muscle cells may undergo DNA synthesis without cell division, leading to a polyploid state. To better understand the nature of smooth muscle cell growth in healthy and pulmonary hypertensive neonatal calves, we measured incorporation of the thymidine analog bromodeoxyuridine (BrdUrd) and total DNA content in medial cells from control (pulmonary arterial pressure = 32 +/- 2 mm of Hg) and hypobaric hypoxia-exposed (pulmonary arterial pressure = 120 +/- 7 mm of Hg) calves. Labeling of medial cells with BrdUrd measured by flow cytometry was increased (P < 0.02) in pulmonary arteries of hypoxia-exposed calves (n = 5), compared with control calves (n = 5). Immunohistochemical localization of BrdUrd indicated that BrdUrd labeling of large elastic pulmonary arteries from hypoxia-exposed calves was increased almost exclusively in the outer half of the medial wall. Increased BrdUrd labeling of muscular pulmonary arteries from hypoxia exposed calves was observed in the arterial media and adventitia, and tended to exit in clusters. Analysis of DNA content by flow cytometry indicated a decrease (P < 0.05) in percentage of tetraploid medial cells in pulmonary arteries from hypoxia-exposed calves, compared with control calves. Bivariate analysis for BrdUrd labeling and DNA content of cells from the pulmonary arteries of hypoxia-exposed calves indicated a subpopulation of diploid cells with positive BrdUrd labeling, suggestive of DNA synthesis and subsequent cell division.(ABSTRACT TRUNCATED AT 250 WORDS)     

多因素法构建肉鸡腹水综合征实验模型及其对肺组织血管内皮舒缩活性因子的影响

试验旨在研究多因素法构建肉鸡腹水综合征（AS）实验模型及其对肺组织血管内皮舒缩活性因子的影响。将158羽健康罗斯肉鸡随机分为空白组、低温模型组和多因素模型组，于25、35、45日龄随机取样，观察临床症状、剖检变化及肺组织病理变化，测定体重、发病率、腹水心脏指数（AHI），检测肺组织一氧化氮（NO）、内皮素-1（ET-1）、一氧化氮合酶（NOS）变化，并以实时荧光定量PCR法检测肺组织内皮素A型受体（ETAR）及血管内皮生长因子（VEGF）mRNA表达量。结果显示，各日龄低温模型组和多因素模型组肉鸡AHI均极显著高于空白组（P<0.01），各日龄多因素模型组肉鸡AHI均显著高于低温模型组（P<0.05）；35、45日龄低温模型组和各日龄多因素模型组肉鸡体重显著或极显著低于空白组（P<0.05，P<0.01）；35、45日龄低温模型组和各日龄多因素模型组AS发病率均极显著高于空白组（P<0.01），25日龄多因素模型组AS发病率极显著高于低温模型组（P<0.01）；35日龄AS肉鸡肺组织结构损伤，内皮细胞脱落、平滑肌排列紊乱；与空白组相比，35、45日龄低温模型组和多因素模型组肺组织NO与ET-1含量、iNOS活性、ETAR及VEGF mRNA表达量均显著或极显著升高（P<0.05，P<0.01），肺组织NO/ET-1、cNOS与TNOS活性均显著或极显著降低（P<0.05，P<0.01）。表明低温、高钠、高能饲料多因素法能够成功构建AS实验模型，多因素造模法较低温造模法更能反映疾病因素的多样性；肺组织血管内皮舒缩活性因子表达紊乱参与了AS的发生发展。     

L-Arginine inhibiting pulmonary vascular remodelling is associated with promotion of apoptosis in pulmonary arterioles smooth muscle cells in broilers

Objective

Pulmonary vascular remodelling is one of the important pathological bases of broiler pulmonary hypertension syndrome (PHS). Nitric oxide (NO) has been found to inhibit proliferation and to induce apoptosis in pulmonary artery smooth muscle cells (SMC) in mammals with pulmonary hypertension. The present study was conducted to evaluate the effects of NO precursor l-arginine on pulmonary vascular remodelling in broilers with pulmonary hypertension induced by cold exposure and to examine whether NO-induced apoptosis in pulmonary arteriole SMC is involved in the regulatory mechanisms.

Methods

Two hundred and forty mixed-sex commercial broilers were equally assigned to three groups and reared in normal brooding temperatures before day 14. Starting on day 14 continuing until the end of the experiment, the control group was brooded in normal temperatures whereas the other two groups were subjected to low ambient temperatures with or without l-arginine added to the basal diets. Cumulative PHS mortality and body weight were recorded in each group. Right/total ventricle ratio (RV/TV), plasma NO concentration and pulmonary vascular morphological changes were analyzed. TdT-mediated dUTP-biotin nick-end labeling (TUNEL) assay was used to detect apoptosis in pulmonary arteriole SMC.

Result

l-Arginine, in group A, had no effect on body weights under cold temperature condition. Birds kept in group B had increased PHS mortality, RV/TV ratio, vessel wall area/vessel total area ratios (WA/TA) and mean media thickness in pulmonary arterioles (mMTPA) (P < 0.05). Percentages of apoptotic SMC in pulmonary arterioles in group B were not altered by cold exposure (P > 0.05). Supplemental dietary l-arginine in group A elevated plasma NO level (P < 0.05), reduced PHS mortality (P < 0.05), attenuated pulmonary vascular remodelling and increased the percentages of apoptotic SMC (P < 0.05) when compared with the group B.

Conclusion

Supplemental l-arginine partially inhibited pulmonary vascular remodelling that occurred secondary to increased pulmonary pressure; NO-induced apoptosis in arteriole SMC might contribute to its regulatory effect on pulmonary vascular structural changes.     

Activation of PKCalpha and pulmonary vascular remodelling in broilers

OBJECTIVE: The present study was conducted to examine the presence of protein kinase Calpha (PKCalpha) in the pulmonary arterioles of broilers during the development of pulmonary hypertension and pulmonary vascular remodelling. METHOD: One hundred and sixty day-old Avian-2000 broilers were divided equally into a control group and a cold temperature group. All the birds were reared in normal temperatures up to day 14, with the lighting schedule at 24 h per day. Thereafter, birds in the cold temperature group were subjected to low temperature by lowering 1-2 degrees C per day to 12-14 degrees C, and then kept constant until day 49, while birds in the control group were still brooded at normal temperatures. All the birds were fed a diet of pellets throughout the study. Samples of blood were taken from the wing vein, and of heart and lung collected after the birds were killed with an overdose of sodium pentobarbitial, at days 24, 32, 39 and 45 of age, respectively. Right ventricle to total ventricle ratio (RV/TV) and packed cell volume (PCV) were measured. Vessel wall area to vessel total area ratio (WA/TA) and mean media thickness in pulmonary arterioles (mMTPA) was examined using computer-image analytic software. Expression of PKC in pulmonary muscular arterioles was assessed by immunohistochemistry and quantified by measuring optical density (OD) using computer-image analytic software. RESULTS: The incidence of pulmonary hypertension syndrome (PHS) was 12.5% in birds exposed to cold, and 3.75% in the control group (P<0.05). PCV in the cold temperature group was elevated after day 32 (P<0.05), and RV/TV ratio increased on day 45 (P<0.05). Both the WA/TA and mMTPA of birds subjected to cold were significantly elevated (P<0.05). The OD values were not significantly increased before day 32 (P>0.05), however, one week later (at day 39 of age), the difference between the two groups was significant (P<0.05). The increased PKCalpha expression was positively correlated with the values of mMTPA and WA/TA. CONCLUSION: PKCalpha expression was up-regulated during the development of pulmonary hypertension. The activation of PKCalpha might be involved in the development of pulmonary vascular remodelling.     

Idiopathic arterial medial calcification of the thoracic arteries in an adult horse

A 6-year-old, gelded, Paint horse displayed clinical signs of muscle wasting and limb stiffness for a 6-month period. The horse's clinical signs abated with corticosteroid therapy, but returned upon cessation of treatment. Upon necropsy, severe lesions of aortic thickening and aortic valve rigidity were observed. Histologically, the tunica media of the aorta, coronary arteries, and pulmonary arteries were expanded by foci of elastin fiber calcification and extracellular matrix with lacunae formation. The vascular lesions are comparative to what has been described as medial arterial calcification, seen in humans suffering from chronic renal failure or diabetes mellitus. No exposure to vitamin D-containing plants or feedstuff could be documented at the time of onset or during the period of clinical signs. The current case describes dramatic lesions of arterial medial calcification of the aorta, coronary, and pulmonary arteries of undetermined cause.     

Trifolium pratense isoflavones improve pulmonary vascular remodelling in broiler chickens

Pulmonary arterial remodelling is a pathological characteristic of pulmonary arterial hypertension (PAH), which contributes to the development of sustained pulmonary hypertension. The aim of this study was to investigate the effects of dietary Trifolium pratense isoflavones on pulmonary vascular remodelling in experimental broiler pulmonary hypertension syndrome. Exposure to sub‐thermoneutral environmental temperatures increased broiler's pulmonary hypertension syndrome incidence and raised expression levels of nitric oxide, endothelin and endothelial nitric oxide synthase. Dietary supplementation (20 mg/kg basal diet) with Trifolium pratense isoflavones reduced pulmonary hypertension syndrome incidence and improved pulmonary vascular remodelling without affecting growth performance. The beneficial effect likely came from isoflavone improved pulmonary vascular remodelling. Isoflavone induced inducible nitric oxide synthase expression, which led to increased nitric oxide level. The nitric oxide could mediate vasorelaxation in the lungs. At the same time, the expression of endothelin was downregulated by isoflavone. Dietary supplementation of Trifolium pratense isoflavone might be a potential therapeutic strategy for the treatment of pulmonary hypertension.     

真武汤对肉鸡腹水综合征的防治作用

选用15日龄艾维茵肉仔鸡256只,随机分成C1、C2、T1、T24个处理组。C1组在常温(18~22℃)环境中饲养,饮水中不添加任何药物,为正常对照组;T1、T2和C2组在低温(8~12℃)环境中饲养。人工复制肉鸡腹水综合征(AS)模型,T1组饮水中按肉鸡体重添加真武汤口服液剂量为0.8 mL.kg-1(中药组);T2组饮水中添加维生素C和维生素E,剂量分别为0.5 g.L-1和0.1 g.L-1,作为药物预防组(维生素组),用药期均为20 d;C2组饮水中不添加任何药物,为模型对照组,以探讨真武汤口服液对肉鸡AS的预防效果。结果表明:整个试验期间,T1、C1组腹水发病率极显著低于C2组(P<0.01),T2组腹水发病率显著低于C2组(P<0.05),T1、C1组发病率最低,为3.13%,C2组发病率最高,为17.19%;C2组腹水心脏指数(AHI)极显著高于C1组(P<0.01),显著高于T1组(P<0.05);T1组血浆、心、肝和肺组织中超氧化物歧化酶(SOD)活性显著高于C2组(P<0.05),同时T1组血浆、心、肝和肺组织中丙二醛(MDA)含量显著低于C2组(P<0.05)。结论:饮水中添加真武汤口服液,能提高机体抗氧化能力,改善血液组织中的理化参数,提高机体抗冷应激能力,明显降低腹水综合征的发生。     

Possible role of histamine in the genesis of pulmonary arterial disease in cats infected with Toxocara cati

Kittens between 12 and 20 weeks of age were orally dosed with 6000 infective ova of Toxocara cati. Animals were sacrificed at intervals between one and eight weeks after infection to study the development of pulmonary arterial lesions. After two weeks, marked leucocyte infiltration and mild thickening of the media of some of the smaller arteries was apparent histologically . Cellular inflammatory activity progressively increased up to four weeks after infection when intimal proliferation was evident in many of the arteries. After six weeks, the arterial walls were grossly thickened with pronounced intimal proliferation which after eight weeks had resulted in complete occlusion of some vessels. The progressive arterial occlusion was associated with a three- to four-fold increase in the parenchymal mast cell population and a corresponding increase in lung histamine content. The possible role of histamine in the genesis of the arteriopathy is discussed.     

高钠所致肺动脉高压肉鸡肺细小动脉病理改变的图象分析

240羽健康AA肉鸡随机均分为试验1组，试验2组和对照组，从8日龄起分别饮用含Na^ 为0.06%、0.12%和0.0%的饮水，其它饲养管理条件相同，分别于12、19、30、34、39日龄抽取各组参试鸡，以右心室（Right ventricle,RV)与全心室（Total ventrcle,TV)的重量比（RV／TV）作为判定肺动脉高压的依据，用图象分析仪对肺细小动脉病理变化作定量检测。结果表明：34、39日龄时，试验1组和试验2组管壁面积／管总面积和中膜厚度占外径百分值明显大于对照组，肺小动脉密度明显低，其RV／TV值均大于0.25，表明试验组肉鸡发生了肺动脉高压。由此可见，由高钠引起肉鸡肺细小动脉血管重构的病变可能参与了肺动脉高压的形成过程。     

Morphologic Changes in the Lungs of Cats Experimentally Infected With Dirofilaria immitis

The morphologic response of the pulmonary arteries and lungs in cats was studied after a five month heartworm infection produced by transplantation of four adult heartworms/cat. One group of seven heartworm infected cats was not treated, another group of seven cats was treated with 97.5 mg of aspirin given twice a week, and the third group of six cats was given aspirin at a sufficient dosage to block in vitro platelet aggregation throughout the study. A fourth group of eight noninfected cats served as controls. Five months after heartworm infection, the cats were euthanized and the lungs perfusion fixed for light microscopy and scanning electron microscopy of the pulmonary arterial surfaces. All cats in the three heartworm-infected groups had live heartworms and the typical pulmonary arterial changes of heartworm disease at necropsy. The arterial surfaces, as viewed with scanning electron microscopy, had villus proliferations that were more numerous and exuberant than similar infections in dogs. Mean percentage of arterial surface involvement with villus proliferation of the nontreated heartworm infected cats was 67.3%; the aspirin treated cats, 73.8%; and the adjusted aspirin treated cats, 75.9%. The villi were myointimal proliferations in the small and medium-sized arteries. The more elastic arteries had a predominance of fibromuscular proliferation. All heartworm infected cats had arterial muscular hypertrophy of the small arteries, in contrast to only three of eight of the nonheartworm infected cats. The caudal lobar arteries were frequently obstructed with either villus proliferation, thrombi, and/or dead heartworms. The muscular arteries had branches with marked dilation, a condition associated with pulmonary hypertension in man. However, only three cats, one in each group, had pulmonary hypertension.(ABSTRACT TRUNCATED AT 250 WORDS)     

Early arterial injury-induced myointimal proliferation in canine pulmonary arteries

Transmission electron microscopy was used to study the early vascular response induced by arterial damage with heartworm infection. The pulmonary arteries were examined in dogs 4 days after experimental transplantation of 6 to 8 Dirofilaria immitis adults. Evan's blue dye was given IV and followed in 60 minutes by perfusion fixation with 1% glutaraldehyde. Endothelial cell junctions were disrupted and rounded endothelial cells were observed. Macrophages and neutrophils adhered to abnormal endothelial cells. Focal areas had platelet aggregates adhered to exposed subendothelial structures. Platelets were activated and degranulated. Areas of the internal elastic lamina appeared to be disrupted, and smooth muscle cell processes from the tunica media extended through these regions of disruption. Smooth muscle cells were oriented toward the arterial lumen and some had migrated to the surface. These findings are compatible with the response to injury theory of myointimal proliferation.     

Arteriosclerosis of extramural coronary arteries in labradors with congestive heart failure

Two adult male labrador dogs had short clinical histories of congestive cardiac failure lasting only six weeks. Cardiac function deteriorated rapidly in both dogs, in spite of symptomatic treatment with cardiac glycoside, diuretic and p-adrenergic blocker. Necropsies revealed ascites and pulmonary congestion, but no macroscopic evidence of valvular or myocardial disease adequate to account for the cardiac dysfunction. Both dogs had extensive stenosis of extramural coronary arteries. This unusual vascular lesion comprised fibroelastic intimal thickening, discontinuity of internal elastic lamella and focal atrophy of medial smooth muscle. It seems likely that cardiac failure in these dogs was associated with myocardial underperfusion caused by extensive non-thrombotic, non-atherogenic stenosis of extramural coronary arteries.     

Expression of PDGF-β receptor in broilers with pulmonary hypertension induced by cold temperature and its association with pulmonary vascular remodeling

Objective

The purpose of the present study was to characterize the relationship between platelet-derived growth factor β receptor (PDGF-β receptor) expression and pulmonary vascular remodeling found in broilers subjected to cold temperature beginning at 14 days of age.

Method

One hundred and sixty-one-day-old mixed-sex Avian-2000 commercial broilers were randomly divided into a normal temperature group (control) and a cold temperature group (cold). All the birds were brooded in normal temperature up to day 14, with the lighting schedule at 24 h per day. Starting at day 14, birds in the cold group were moved to a pen in the cold house and subjected to low temperature, while birds in the control group were still brooded at normal temperature. On days 14, 23, 30, 37 and 44, the right/total ventricle weight ratio (RV/TV), packed cell volume (PCV), the vessel wall area to vessel total area ratio (WA/TA), mean media thickness in pulmonary arterioles (mMTPA) and the expression of PDGF-β receptor in pulmonary arterioles were measured, respectively. Cumulative pulmonary hypertension syndrome (PHS) morbidity was recorded in each group.

Results

Cool ambient temperature increased PHS morbidity of broilers. The values of WA/TA and mMTPA were also increased significantly compared with control group. PCV values in the cold temperature group were elevated from days 30 to 44, and RV/TV ratios were increased on days 37 and 44. Cold exposure enhanced PDGF-β receptor expression in pulmonary arterioles, and the PDGF-β receptor expression was significantly correlated with pulmonary vascular remodeling that was dedicated by increased WA/TA and mMTPA.

Conclusion

The results indicated that PDGF-β and its receptor were involved in the underlying mechanisms of pulmonary vascular remodeling in pulmonary hypertensive broilers.     

参芪颗粒预防肉鸡腹水综合征的研究

15日龄雄性AA肉仔鸡240只,分为6组,一组于常规温度下饲养,为正常对照组;其余5组均在低温下饲养,一组为硝苯地平组,为药物对照组;一组不给任何药物,为阳性对照组;余下3组作为试验组,分别给予高、中、低剂量的参芪颗粒制剂,连用15 d。结果表明:低温饲养能明显提高肉鸡腹水发病率,中剂量及高剂量组腹水发病率明显低于阳性对照组。腹水心脏指数、肺动脉压指标显示,药物对照组、不同剂量参芪颗粒组均能不同程度地改善腹水心脏指数及肺动脉压,参芪颗粒对肉鸡腹水综合征有防治作用。