HP-PRRSV抑制猪肺eNOS表达与猪肺微血栓形成的相关性研究

doi:10.11843/j.issn.0366-6964.2020.04.020

畜牧兽医学报 ›› 2020, Vol. 51 ›› Issue (4): 834-840.doi: 10.11843/j.issn.0366-6964.2020.04.020

HP-PRRSV抑制猪肺eNOS表达与猪肺微血栓形成的相关性研究

赵红利, 罗传真, 娄江城, 陈梦月, 刘晓丽, 谷长勤, 张万坡, 程国富, 刘榜, 胡薛英*

华中农业大学动物科学技术学院-动物医学院, 武汉 430070

收稿日期:2019-10-08 出版日期:2020-04-25 发布日期:2020-04-21
通讯作者: 胡薛英,主要从事兽医病理学研究,E-mail:hxying@mail.hzau.edu.cn
作者简介:赵红利(1994-),女,陕西西安人,硕士生,主要从事兽医病理学研究,E-mail:1105848824@qq.com
基金资助:
国家自然科学基金重点项目（31930104）

Study on the Relationship between the Inhibition of eNOS Expression by HP-PRRSV and the Formation of Microthrombus in Pig Lung

ZHAO Hongli, LUO Chuanzhen, LOU Jiangcheng, CHEN Mengyue, LIU Xiaoli, GU Changqin, ZHANG Wanpo, CHENG Guofu, LIU Bang, HU Xueying*

College of Veterinary Medicine, Huazhong Agricultural University, Wuhan 430070, China

Received:2019-10-08 Online:2020-04-25 Published:2020-04-21

摘要/Abstract

摘要： 为了解高致病性猪繁殖与呼吸综合征病毒（HP-PRRSV）与肺微血栓病变的关系及内皮型一氧化氮合酶（eNOS）的表达对微血栓的影响，选取50头70日龄的鄂通两头乌猪，4头作为正常对照，46头感染HP-PRRSV。采取50头猪的肺组织，应用HE染色观察组织病理变化并对微血栓病变程度进行等级评分，将46头感染猪分为轻度微血栓组、中度微血栓组、重度微血栓组，每组选取4头猪进行后续研究。应用免疫组化、荧光定量PCR和Western blot技术，明确HP-PRRSV和eNOS在微血栓猪肺中的表达、分布及变化。组织病理学研究结果显示感染猪肺组织肺泡间隔明显增宽，毛细血管有不同程度的淤血、出血，肺泡腔内有巨噬细胞、脱落的肺泡上皮细胞及淋巴细胞，肺泡壁有大量微血栓。HP-PRRSV主要分布于肺泡巨噬细胞及少量的淋巴细胞的细胞质。随着HP-PRRSV病毒含量的增加，微血栓病变越严重。微血栓病变越严重，死亡率越高。eNOS主要分布于肺泡Ⅱ型细胞、巨噬细胞、血管内皮细胞及支气管平滑肌细胞的细胞质，感染猪肺组织的eNOS mRNA和蛋白表达显著低于未感染对照组。研究结果表明HP-PRRSV使eNOS的表达下调，并参与感染猪肺组织微血栓的形成，增加感染猪的死亡率。

关键词: 高致病性猪繁殖与呼吸综合征病毒, 鄂通两头乌猪, 肺, 微血栓, 内皮型一氧化氮合酶

Abstract: There are two objectives of the study. First is to explore the relationship between HP-porcine reproductive and respiratory syndrome virus (HP-PRRSV) and pathological changes of pulmonary microthrombus, the other is to clarify the effect of endothelial nitric oxide synthase (eNOS) expression on thrombosis. Fifty 70-day-old healthy piglets were employed, control group consisted of four piglets and 46 piglets were infected with HP-PRRSV. Lungs were collected, HE technique was then applied to observe histopathological changes and to evaluate thrombus grade. Mild thrombus group, moderate thrombus group and severe thrombus group were assigned based on state of their thromboses according to the statistics, four piglets in each group were selected for follow-up study. Immunohistochemistry, Western blot, real-time PCR were employed in examining the distribution, expression and changes of HP-PRRSV and eNOS in the lungs of piglets. The results of histopathological studies showed that the alveolar space of the lung tissue was significantly widened, different degree of blood stasis and bleeding in blood capillary occurred, alveolar epithelial cells, macrophages peeled off in the alveolar cavity and amounts of microthrombus in the alveolar wall were also observed. HP-PRRSV was found mainly located in cytoplasm of alveolar macrophages and fewer in lymphocytes. The degree of thrombosis mounted associated with the increase of viral content and then lead to a higher mortality rate. In addition, eNOS was found mainly located in the cytoplasm of alveolar type Ⅱ cells, macrophages, vascular endothelial cells and bronchial smooth muscle cells. The expression of eNOS mRNA and protein in infected pig lungs was significantly lower compared with controls but there was no obvious difference between the three infected groups. In summary, HP-PRRSV develops the mortality of infected pigs by reducing the expression of eNOS and participated in the formation of microthrombosis.

Key words: HP-PRRSV, Etongliangtouwu pig, lung, microthrombus, eNOS

中图分类号:

S858.285.3

赵红利, 罗传真, 娄江城, 陈梦月, 刘晓丽, 谷长勤, 张万坡, 程国富, 刘榜, 胡薛英. HP-PRRSV抑制猪肺eNOS表达与猪肺微血栓形成的相关性研究[J]. 畜牧兽医学报, 2020, 51(4): 834-840.

ZHAO Hongli, LUO Chuanzhen, LOU Jiangcheng, CHEN Mengyue, LIU Xiaoli, GU Changqin, ZHANG Wanpo, CHENG Guofu, LIU Bang, HU Xueying. Study on the Relationship between the Inhibition of eNOS Expression by HP-PRRSV and the Formation of Microthrombus in Pig Lung[J]. Acta Veterinaria et Zootechnica Sinica, 2020, 51(4): 834-840.

参考文献 14

[1]	LUNNEY J K, BENFIELD D A, ROWLAND R R R. Porcine reproductive and respiratory syndrome virus:an update on an emerging and re-emerging viral disease of swine[J]. Virus Res, 2010, 154(1-2):1-6.
[2]	WAGNER J, KNEUCKER A, LIEBLER-TENORIO E, et al. Respiratory function and pulmonary lesions in pigs infected with porcine reproductive and respiratory syndrome virus[J]. Vet J, 2011, 187(3):310-319.
[3]	佘锐萍. 动物病理学[M]. 北京:中国农业出版社, 2007:72-75.SHE R P. Animal pathology[M]. Beijing:China Agriculture Press, 2007:72-75.
[4]	刘昭娅, 胥茜, 唐义信, 等. 内皮型一氧化氮合酶活性的调节与心血管疾病的研究进展[J]. 中南大学学报:医学版, 2016, 41(6):632-636.LIU Z Y, XU Q, TANG Y X, et al. Research progress in the regulation of endothelial nitric oxide synthase activity and cardiovascular diseases[J]. Journal of Central South University:Medical Science, 2016, 41(6):632-636. (in Chinese)
[5]	GIELIS J F, LIN J Y, WINGLER K, et al. Pathogenetic role of eNOS uncoupling in cardiopulmonary disorders[J]. Free Radical Biol Med, 2011, 50(7):765-776.
[6]	徐三平, 刘榜, 吴招甫, 等. 以通城猪为素材的新品种培育——"鄂通两头乌"[J]. 中国猪业, 2014(11):66-68.XU S P, LIU B, WU Z F, et al. Cultivation of new breed with Tongcheng pig as material-"etongliangtouwu"[J]. China Swine Industry, 2014(11):66-68. (in Chinese)
[7]	宋爽. PRRSV诱导炎症反应及其调控机制[D]. 武汉:华中农业大学, 2013.SONG S. PRRSV-induced inflammatory response and its regulatory mechanism[D]. Wuhan:Huazhong Agricultural University, 2013. (in Chinese)
[8]	翟研妮. 大白猪和通城猪感染HP-PRRSV后比较组织病理学研究[D]. 武汉:华中农业大学, 2012.ZHAI Y N. Comparative histological and pathologic study on large-white and Tongcheng pigs that infected HP-PRRSV[D]. Wuhan:Huazhong Agricultural University, 2012. (in Chinese)
[9]	KRAUSE B J, CARRASCO-WONG I, CANIUGUIR A, et al. Endothelial eNOS/arginase imbalance contributes to vascular dysfunction in IUGR umbilical and placental vessels[J]. Placenta, 2013, 34(1):20-28.
[10]	刘玲燕, 兰茜, 朱庆. 内皮一氧化氮合酶(eNOS)基因与藏鸡低氧适应机制的研究[J].畜牧兽医学报,2015,46(2):192-195.LIU L Y,LAN X,ZHU Q. The research of endothelial nitric oxide synthase gene in hypoxia adaptation of Tibetan chicken[J]. Acta Veterinaria et Zootechnica Sinica, 2015, 46(2):192-195. (in Chinese)
[11]	KOLLURU G K, SIAMWALA J H, CHATTERJEE S. eNOS phosphorylation in health and disease[J]. Biochimie, 2010, 92(9):1186-1198.
[12]	FÖRSTERMANN U, SESSA W C. Nitric oxide synthases:regulation and function[J]. Eur Heart J, 2012, 33(7):829-837.
[13]	KRAM L, GRAMBOW E, MUELLER-GRAF F, et al. The anti-thrombotic effect of hydrogen sulfide is partly mediated by an upregulation of nitric oxide synthases[J]. Thromb Res, 2013, 132(2):e112-e117.
[14]	SOOD B G, WYKES S, LANDA M, et al. Expression of eNOS in the lungs of neonates with pulmonary hypertension[J]. Exp Mol Pathol, 2011, 90(1):9-12.

[1]	王鑫鑫, 林树梅, 赵冬冬, 王学生. 肺泡上皮细胞分泌的外泌体调控巨噬细胞极化在急性肺损伤中的作用[J]. 畜牧兽医学报, 2024, 55(1): 71-78.
[2]	张颖, 袁莉刚, 陈国娟, 张芳, 杨大鹏. ET-1/eNOS表达差异在牦牛隐睾发生中的作用分析[J]. 畜牧兽医学报, 2024, 55(1): 207-217.
[3]	谢青云, 邢蕙萱, 于岩飞, 袁厅, 熊祺琰, 熊富强, 冯志新. 猪肺炎支原体解螺旋酶RuvA的原核表达、多克隆抗体制备及活性鉴定[J]. 畜牧兽医学报, 2024, 55(1): 271-281.
[4]	巩志国, 赵佳敏, 顾柏臣, 任佩佩, 于琢雅, 白云洁, 刘鑫煜, 王超, 刘博. 基于网络药理学分析党参减轻大肠杆菌感染小鼠急性肺损伤的作用机制[J]. 畜牧兽医学报, 2023, 54(8): 3571-3581.
[5]	赵菲菲, 李杰, 韩宁, 谢仕廷, 曾振灵. 分离自屠宰场的肺炎克雷伯菌的耐药性分析[J]. 畜牧兽医学报, 2023, 54(7): 3044-3053.
[6]	曹西月, 纪晓霞, 张崇昊, 张亚峰, 陈雨涛, 张源淑. 二脒那秦通过内源性激活ACE2抑制AngⅡ-TGF-β1通路缓解小鼠肺纤维化[J]. 畜牧兽医学报, 2023, 54(6): 2631-2640.
[7]	陆梦琪, 杨文杰, 李萍, 余鹏, 董翎, 牛晓玉, 杨克礼, 邹维华, 宋卉. 基于16S rRNA测序分析PRRSV感染仔猪肺和肠道中微生物菌群的变化[J]. 畜牧兽医学报, 2023, 54(4): 1664-1678.
[8]	陈永平, 寇玉红, 焦文静, 侯晓昱, 范宏刚. 辅酶Q10改善LPS诱导小鼠急性肺损伤的效应分析[J]. 畜牧兽医学报, 2023, 54(4): 1730-1741.
[9]	卢碧凯, 袁秀芳, 徐丽华, 余斌, 苏菲, 叶十一, 陈怡洁, 蒋利明, 张晖, 李军星. 胸膜肺炎放线杆菌分离株血清型的PCR鉴定和Apx毒素基因检测[J]. 畜牧兽医学报, 2023, 54(3): 1341-1346.
[10]	徐婷婷, 齐芬芳, 黄世会, 牛熙, 李升, 冉雪琴, 王嘉福, 谢健. 香猪MAP3K4基因结构变异多态性和基因表达研究[J]. 畜牧兽医学报, 2023, 54(12): 5046-5055.
[11]	孟令宅, 陈春丽, 于蒙蒙, 王占新, 王素艳, 刘鹏, 何塔娜, 郭茹, 陈运通, 刘长军, 祁小乐, 吴志强, 高玉龙. B亚型禽偏肺病毒对黄羽肉鸡致病性分析及其灭活疫苗免疫效果评价[J]. 畜牧兽医学报, 2023, 54(12): 5154-5161.
[12]	王佳宁, 张自强, 孔德婧, 冯彩彩, 张飞可, 刘玉梅. 家兔肺炎克雷伯菌的分离鉴定[J]. 畜牧兽医学报, 2023, 54(12): 5198-5206.
[13]	张凯川, 王晋宇, 李守军, 贾坤. 广东省羊源肺炎克雷伯菌遗传进化与毒力基因及耐药性分析[J]. 畜牧兽医学报, 2023, 54(1): 328-337.
[14]	王剑, 陈雪苹, 李继昌. 唾液乳杆菌对鸡毒支原体感染肉鸡生长性能及肺损伤的影响[J]. 畜牧兽医学报, 2022, 53(9): 3172-3179.
[15]	仝锡帅, 余耿生, 李雅文, 冉迪, 顾建红, 邹辉, 袁燕, 卞建春, 刘宗平. NAC通过抑制大鼠肺细胞凋亡缓解镉致肺组织损伤[J]. 畜牧兽医学报, 2022, 53(9): 3241-3250.

HP-PRRSV抑制猪肺eNOS表达与猪肺微血栓形成的相关性研究

Study on the Relationship between the Inhibition of eNOS Expression by HP-PRRSV and the Formation of Microthrombus in Pig Lung

RichHTML

PDF

可视化

摘要/Abstract

引用本文

使用本文

参考文献 14

相关文章 15

编辑推荐

Metrics

本文评价