{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2025,10,11]],"date-time":"2025-10-11T02:18:36Z","timestamp":1760149116245,"version":"build-2065373602"},"reference-count":27,"publisher":"MDPI AG","issue":"7","license":[{"start":{"date-parts":[[2023,7,8]],"date-time":"2023-07-08T00:00:00Z","timestamp":1688774400000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"name":"National Natural Science Foundation of China","award":["32273050","31672597","6202003","5212003"],"award-info":[{"award-number":["32273050","31672597","6202003","5212003"]}]},{"name":"Beijing Municipal Natural Science","award":["32273050","31672597","6202003","5212003"],"award-info":[{"award-number":["32273050","31672597","6202003","5212003"]}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Viruses"],"abstract":"<jats:p>The highly pathogenic porcine reproductive and respiratory syndrome virus (HP-PRRSV) often causes secondary bacterial infection in piglets, resulting in inflammatory lung injury and leading to high mortality rates and significant economic losses in the pig industry. Microvascular endothelial cells (MVECs) play a crucial role in the inflammatory response. Previous studies have shown that HP-PRRSV can infect porcine pulmonary MVECs and damage the endothelial glycocalyx. To further understand the role of pulmonary MVECs in the pathogenesis of HP-PRRSV and its secondary bacterial infection, in this study, cultured porcine pulmonary MVECs were stimulated with a HP-PRRSV HN strain and lipopolysaccharide (LPS). The changes in gene expression profiles were analyzed through transcriptome sequencing, and the differentially expressed genes were verified using qRT-PCR, Western blot, and ELISA. Furthermore, the effects on endothelial barrier function and regulation of neutrophil trans-endothelial migration were detected using the Transwell model. HP-PRRSV primarily induced differential expression of numerous genes associated with immune response, including IFIT2, IFIT3, VCAM1, ITGB4, and CCL5, whereas LPS triggered an inflammatory response involving IL6, IL16, CXCL8, CXCL14, and ITGA7. Compared to the individual effect of LPS, when given after HN-induced stimulation, it caused a greater number of changes in inflammatory molecules, such as VCAM1, IL1A, IL6, IL16, IL17D, CCL5, ITGAV, IGTB8, and TNFAIP3A, a more significant reduction in transendothelial electrical resistance, and higher increase in neutrophil transendothelial migration. In summary, these results suggest a synergistic effect of HP-PRRSV and LPS on the inflammatory response of porcine pulmonary MVECs. This study provides insights into the mechanism of severe lung injury caused by secondary bacterial infection following HP-PRRSV infection from the perspective of MVECs, emphasizing the vital role of pulmonary MVECs in HP-PRRSV infection.<\/jats:p>","DOI":"10.3390\/v15071523","type":"journal-article","created":{"date-parts":[[2023,7,10]],"date-time":"2023-07-10T00:45:37Z","timestamp":1688949937000},"page":"1523","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":2,"title":["Synergistic Effect of Treatment with Highly Pathogenic Porcine Reproductive and Respiratory Syndrome Virus and Lipopolysaccharide on the Inflammatory Response of Porcine Pulmonary Microvascular Endothelial Cells"],"prefix":"10.3390","volume":"15","author":[{"given":"Xinyue","family":"Yao","sequence":"first","affiliation":[{"name":"Beijing Key Laboratory of Traditional Chinese Veterinary Medicine, Animal Science and Technology College, Beijing University of Agriculture, No. 7 Beinong Road, Beijing 102206, China"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Wanwan","family":"Dai","sequence":"additional","affiliation":[{"name":"College of Veterinary Medicine, Shanxi Agriculture University, Taigu 030801, China"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Siyu","family":"Yang","sequence":"additional","affiliation":[{"name":"Beijing Key Laboratory of Traditional Chinese Veterinary Medicine, Animal Science and Technology College, Beijing University of Agriculture, No. 7 Beinong Road, Beijing 102206, China"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Zhaoli","family":"Wang","sequence":"additional","affiliation":[{"name":"Beijing Key Laboratory of Traditional Chinese Veterinary Medicine, Animal Science and Technology College, Beijing University of Agriculture, No. 7 Beinong Road, Beijing 102206, China"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Qian","family":"Zhang","sequence":"additional","affiliation":[{"name":"Beijing Key Laboratory of Traditional Chinese Veterinary Medicine, Animal Science and Technology College, Beijing University of Agriculture, No. 7 Beinong Road, Beijing 102206, China"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Qinghui","family":"Meng","sequence":"additional","affiliation":[{"name":"Beijing Milu Ecological Research Center, Beijing Research Institute of Science and Technology, Beijing 100076, China"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-2631-158X","authenticated-orcid":false,"given":"Tao","family":"Zhang","sequence":"additional","affiliation":[{"name":"Beijing Key Laboratory of Traditional Chinese Veterinary Medicine, Animal Science and Technology College, Beijing University of Agriculture, No. 7 Beinong Road, Beijing 102206, China"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"1968","published-online":{"date-parts":[[2023,7,8]]},"reference":[{"key":"ref_1","first-page":"11","article-title":"PRRSV, the virus","volume":"31","author":"Meulenberg","year":"2000","journal-title":"Vet. Res."},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"30","DOI":"10.1016\/j.vetmic.2017.02.020","article-title":"Pathogenesis and control of the Chinese highly pathogenic porcine reproductive and respiratory syndrome virus","volume":"209","author":"Han","year":"2017","journal-title":"Vet. Microbiol."},{"doi-asserted-by":"crossref","unstructured":"Li, Z., He, Y., Xu, X., Leng, X., Li, S., Wen, Y., Wang, F., Xia, M., Cheng, S., and Wu, H. (2016). Pathological and immunological characteristics of piglets infected experimentally with a HP-PRRSV TJ strain. BMC Vet. Res., 12.","key":"ref_3","DOI":"10.1186\/s12917-016-0854-x"},{"key":"ref_4","doi-asserted-by":"crossref","first-page":"e2","DOI":"10.4142\/jvs.21139","article-title":"Transcriptome profiling identifies immune response genes against porcine reproductive and respiratory syndrome virus and Haemophilus parasuis co-infection in the lungs of piglets","volume":"23","author":"Zhang","year":"2022","journal-title":"J. Vet. Sci."},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"104536","DOI":"10.1016\/j.micpath.2020.104536","article-title":"High incidence and characteristic of PRRSV and resistant bacterial Co-Infection in pig farms","volume":"149","author":"Xiangjin","year":"2020","journal-title":"Microb. Pathog."},{"key":"ref_6","first-page":"49","article-title":"Respiratory health status in swine producers relates to endotoxin exposure in the presence of low dust levels","volume":"36","author":"Zejda","year":"1994","journal-title":"J. Occup. Med."},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"213","DOI":"10.1016\/j.vetmic.2010.11.006","article-title":"Porcine reproductive and respiratory syndrome virus and bacterial endotoxin act in synergy to amplify the inflammatory response of infected macrophages","volume":"149","author":"Qiao","year":"2011","journal-title":"Vet. Microbiol."},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"1","DOI":"10.1016\/S0378-1135(02)00104-9","article-title":"Porcine reproductive-respiratory syndrome virus infection predisposes pigs for respiratory signs upon exposure to bacterial lipopolysaccharide","volume":"88","author":"Labarque","year":"2002","journal-title":"Vet. Microbiol."},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"535","DOI":"10.1016\/j.ajpath.2019.11.004","article-title":"Endothelial Cell Calcium Signaling during Barrier Function and Inflammation","volume":"190","author":"Dalal","year":"2020","journal-title":"Am. J. Pathol."},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"1417","DOI":"10.1016\/S0140-6736(20)30937-5","article-title":"Endothelial cell infection and endotheliitis in COVID-19","volume":"395","author":"Varga","year":"2020","journal-title":"Lancet"},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"1742528","DOI":"10.1080\/20002297.2020.1742528","article-title":"Porphyromonas gingivalis W83 traffics via ICAM1 in microvascular endothelial cells and alters capillary organization in vivo","volume":"12","author":"Reyes","year":"2020","journal-title":"J. Oral Microbiol."},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"1404","DOI":"10.1292\/jvms.20-0324","article-title":"Susceptibility of porcine pulmonary microvascular endothelial cells to porcine reproductive and respiratory syndrome virus","volume":"82","author":"Li","year":"2020","journal-title":"J. Vet. Med. Sci."},{"doi-asserted-by":"crossref","unstructured":"Song, X., Wu, Y., Wu, X., Hu, G., and Zhang, T. (2022). Effects of highly pathogenic porcine reproductive and respiratory syndrome virus infection on the surface glycoprofiling of porcine pulmonary microvascular endothelial cells. Viruses, 14.","key":"ref_13","DOI":"10.3390\/v14112569"},{"doi-asserted-by":"crossref","unstructured":"Wu, Y., Song, X., Cui, D., and Zhang, T. (2022). IFIT3 and IFIT5 play potential roles in innate immune response of porcine pulmonary microvascular endothelial cells to highly pathogenic porcine reproductive and respiratory syndrome virus. Viruses, 14.","key":"ref_14","DOI":"10.3390\/v14091919"},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"113296","DOI":"10.1016\/j.jim.2022.113296","article-title":"Isolation and culture of rat intestinal mucosal microvascular endothelial cells using immunomagnetic beads","volume":"507","author":"Wu","year":"2022","journal-title":"J. Immunol. Methods"},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"289","DOI":"10.1111\/j.2517-6161.1995.tb02031.x","article-title":"Controlling the False Discovery Rate: A Practical and Powerful Approach to Multiple Testing","volume":"57","author":"Benjamini","year":"1995","journal-title":"J. R. Stat. Soc. Ser. B Stat. Methodol."},{"key":"ref_17","first-page":"293","article-title":"Endothelial activation and dysfunction in COVID-19: From basic mechanisms to potential therapeutic approaches. Signal Transduct","volume":"5","author":"Jin","year":"2020","journal-title":"Target. Ther."},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"606","DOI":"10.1111\/ahe.12521","article-title":"Endothelial-platelet interactions in influenza-induced pneumonia: A potential therapeutic target","volume":"49","author":"Rommel","year":"2020","journal-title":"Anat. Histol. Embryol."},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"102763","DOI":"10.1016\/j.autrev.2021.102763","article-title":"Interleukin 1\u03b1: A comprehensive review on the role of IL-1\u03b1 in the pathogenesis and treatment of autoimmune and inflammatory diseases","volume":"20","author":"Lopalco","year":"2021","journal-title":"Autoimmun. Rev."},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"593","DOI":"10.1586\/1744666X.2014.894886","article-title":"The CXCL8\/IL-8 chemokine family and its receptors in inflammatory diseases","volume":"10","author":"Russo","year":"2014","journal-title":"Expert Rev. Clin. Immunol."},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"166170","DOI":"10.1016\/j.bbadis.2021.166170","article-title":"VCAM-1 as a predictor biomarker in cardiovascular disease","volume":"1867","author":"Troncoso","year":"2021","journal-title":"Biochim. Biophys. Acta Mol. Basis Dis."},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"110012","DOI":"10.1016\/j.intimp.2023.110012","article-title":"Highly pathogenic porcine reproductive and respiratory syndrome virus-induced inflammatory response in porcine pulmonary microvascular endothelial cells and effects of herbal ingredients on main inflammatory molecules","volume":"118","author":"Wu","year":"2023","journal-title":"Int. Immunopharmacol."},{"doi-asserted-by":"crossref","unstructured":"De Lamache, D.D., Moges, R., Siddiq, A., Allain, T., Feener, T.D., Muench, G.P., McKenna, N., Yates, R.M., and Buret, A.G. (2019). Immuno-modulating properties of Tulathromycin in porcine monocyte-derived macrophages infected with porcine reproductive and respiratory syndrome virus. PLoS ONE, 14.","key":"ref_23","DOI":"10.1371\/journal.pone.0221560"},{"key":"ref_24","doi-asserted-by":"crossref","first-page":"137","DOI":"10.1016\/j.vetimm.2015.02.003","article-title":"A comparative study of the local cytokine response in the lungs of pigs experimentally infected with different PRRSV-1 strains: Upregulation of IL-1\u03b1 in highly pathogenic strain induced lesions","volume":"164","author":"Amarilla","year":"2015","journal-title":"Vet. Immunol. Immunopathol."},{"key":"ref_25","doi-asserted-by":"crossref","first-page":"107","DOI":"10.1177\/2211068214561025","article-title":"TEER measurement techniques for in vitro barrier model systems","volume":"20","author":"Srinivasan","year":"2015","journal-title":"J. Lab. Autom."},{"key":"ref_26","doi-asserted-by":"crossref","first-page":"26","DOI":"10.1111\/cns.13560","article-title":"Endothelial glycocalyx as an important factor in composition of blood-brain barrier","volume":"27","author":"Zhao","year":"2021","journal-title":"CNS Neurosci. Ther."},{"key":"ref_27","first-page":"2149","article-title":"Neutrophil transendothelial migration: Updates and new perspectives","volume":"133","author":"Filippi","year":"2019","journal-title":"Blood J. Am. Soc. Hematol."}],"container-title":["Viruses"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/1999-4915\/15\/7\/1523\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,10]],"date-time":"2025-10-10T20:09:07Z","timestamp":1760126947000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/1999-4915\/15\/7\/1523"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2023,7,8]]},"references-count":27,"journal-issue":{"issue":"7","published-online":{"date-parts":[[2023,7]]}},"alternative-id":["v15071523"],"URL":"https:\/\/doi.org\/10.3390\/v15071523","relation":{},"ISSN":["1999-4915"],"issn-type":[{"type":"electronic","value":"1999-4915"}],"subject":[],"published":{"date-parts":[[2023,7,8]]}}}