PMID- 32649510 OWN - NLM STAT- MEDLINE DCOM- 20210514 LR - 20210514 IS - 2542-5641 (Electronic) IS - 0366-6999 (Print) IS - 0366-6999 (Linking) VI - 133 IP - 15 DP - 2020 Aug 5 TI - Unfractionated heparin attenuates endothelial barrier dysfunction via the phosphatidylinositol-3 kinase/serine/threonine kinase/nuclear factor kappa-B pathway. PG - 1815-1823 LID - 10.1097/CM9.0000000000000905 [doi] AB - BACKGROUND: Vascular endothelial dysfunction is considered a key pathophysiologic process for the development of acute lung injury. In this study, we aimed at investigating the effects of unfractionated heparin (UFH) on the lipopolysaccharide (LPS)-induced changes of vascular endothelial-cadherin (VE-cadherin) and the potential underlying mechanisms. METHODS: Male C57BL/6 J mice were randomized into three groups: vehicle, LPS, and LPS + UFH groups. Intraperitoneal injection of 30 mg/kg LPS was used to induce sepsis. Mice in the LPS + UFH group received subcutaneous injection of 8 U UFH 0.5 h before LPS injection. The lung tissue of the mice was collected for assessing lung injury by measuring the lung wet/dry (W/D) weight ratio and observing histological changes. Human pulmonary microvascular endothelial cells (HPMECs) were cultured and used to analyze the effects of UFH on LPS- or tumor necrosis factor-alpha (TNF-alpha)-induced vascular hyperpermeability, membrane expression of VE-cadherin, p120-catenin, and phosphorylated myosin light chain (p-MLC), and F-actin remodeling, and on the LPS-induced activation of the phosphatidylinositol-3 kinase (PI3K)/serine/threonine kinase (Akt)/nuclear factor kappa-B (NF-kappaB) signaling pathway. RESULTS: In vivo, UFH pretreatment significantly attenuated LPS-induced pulmonary histopathological changes (neutrophil infiltration and erythrocyte effusion, alveolus pulmonis collapse, and thicker septum), decreased the lung W/D, and increased protein concentration (LPS vs. LPS + UFH: 0.57 +/- 0.04 vs. 0.32 +/- 0.04 mg/mL, P = 0.0092), total cell count (LPS vs. LPS + UFH: 9.57 +/- 1.23 vs. 3.65 +/- 0.78 x 10/mL, P = 0.0155), polymorphonuclear neutrophil percentage (LPS vs. LPS + UFH: 88.05% +/- 2.88% vs. 22.20% +/- 3.92%, P = 0.0002), and TNF-alpha (460.33 +/- 23.48 vs. 189.33 +/- 14.19 pg/mL, P = 0.0006) in the bronchoalveolar lavage fluid. In vitro, UFH pre-treatment prevented the LPS-induced decrease in the membrane expression of VE-cadherin (LPS vs. LPS + UFH: 0.368 +/- 0.044 vs. 0.716 +/- 0.064, P = 0.0114) and p120-catenin (LPS vs. LPS + UFH: 0.208 +/- 0.018 vs. 0.924 +/- 0.092, P = 0.0016), and the LPS-induced increase in the expression of p-MLC (LPS vs. LPS + UFH: 0.972 +/- 0.092 vs. 0.293 +/- 0.025, P = 0.0021). Furthermore, UFH attenuated LPS- and TNF-alpha-induced hyperpermeability of HPMECs (LPS vs. LPS + UFH: 8.90 +/- 0.66 vs. 15.84 +/- 1.09 Omega.cm, P = 0.0056; TNF-alpha vs. TNF-alpha + UFH: 11.28 +/- 0.64 vs. 18.15 +/- 0.98 Omega.cm, P = 0.0042) and F-actin remodeling (LPS vs. LPS + UFH: 56.25 +/- 1.51 vs. 39.70 +/- 1.98, P = 0.0027; TNF-alpha vs. TNF-alpha + UFH: 55.42 +/- 1.42 vs. 36.51 +/- 1.20, P = 0.0005) in vitro. Additionally, UFH decreased the phosphorylation of Akt (LPS vs. LPS + UFH: 0.977 +/- 0.081 vs. 0.466 +/- 0.035, P = 0.0045) and I kappa B Kinase (IKK) (LPS vs. LPS + UFH: 1.023 +/- 0.070 vs. 0.578 +/- 0.044, P = 0.0060), and the nuclear translocation of NF-kappaB (LPS vs. LPS + UFH: 1.003 +/- 0.077 vs. 0.503 +/- 0.065, P = 0.0078) in HPMECs, which was similar to the effect of the PI3K inhibitor, wortmannin. CONCLUSIONS: The protective effect of UFH against LPS-induced pulmonary endothelial barrier dysfunction involves VE-cadherin stabilization and PI3K/Akt/NF-kappaB signaling. FAU - Mu, Sheng-Tian AU - Mu ST AD - Department of Intensive Care Unit, Cancer Hospital of China Medical University, Liaoning Cancer Hospital and Institute, Shenyang, Liaoning 110042, China. FAU - Tang, Jie AU - Tang J AD - Department of Intensive Care Unit, Cancer Hospital of China Medical University, Liaoning Cancer Hospital and Institute, Shenyang, Liaoning 110042, China. FAU - Ma, Jian-Qi AU - Ma JQ AD - Department of Intensive Care Unit, Cancer Hospital of China Medical University, Liaoning Cancer Hospital and Institute, Shenyang, Liaoning 110042, China. FAU - Zhong, Yu AU - Zhong Y AD - Department of Intensive Care Unit, Cancer Hospital of China Medical University, Liaoning Cancer Hospital and Institute, Shenyang, Liaoning 110042, China. FAU - Liu, Han-Zhe AU - Liu HZ AD - Department of Intensive Care Unit, Cancer Hospital of China Medical University, Liaoning Cancer Hospital and Institute, Shenyang, Liaoning 110042, China. FAU - Ma, Xiao-Chun AU - Ma XC AD - Department of Intensive Care Unit, The First Affiliated Hospital of China Medical University, Shenyang, Liaoning 110001, China. FAU - Zheng, Zhen AU - Zheng Z AD - Department of Intensive Care Unit, Cancer Hospital of China Medical University, Liaoning Cancer Hospital and Institute, Shenyang, Liaoning 110042, China. LA - eng PT - Journal Article PL - China TA - Chin Med J (Engl) JT - Chinese medical journal JID - 7513795 RN - 0 (Lipopolysaccharides) RN - 0 (NF-kappa B) RN - 0 (Phosphatidylinositols) RN - 452VLY9402 (Serine) RN - 9005-49-6 (Heparin) RN - EC 2.7.1.137 (Phosphatidylinositol 3-Kinase) SB - IM MH - Animals MH - Endothelial Cells MH - *Heparin MH - Lipopolysaccharides/toxicity MH - Lung MH - Male MH - Mice MH - Mice, Inbred C57BL MH - *NF-kappa B MH - Phosphatidylinositol 3-Kinase MH - Phosphatidylinositol 3-Kinases MH - Phosphatidylinositols MH - Serine PMC - PMC7470014 COIS- None. EDAT- 2020/07/11 06:00 MHDA- 2021/05/15 06:00 PMCR- 2020/08/05 CRDT- 2020/07/11 06:00 PHST- 2020/07/11 06:00 [pubmed] PHST- 2021/05/15 06:00 [medline] PHST- 2020/07/11 06:00 [entrez] PHST- 2020/08/05 00:00 [pmc-release] AID - 00029330-202008050-00007 [pii] AID - CMJ-2019-1949 [pii] AID - 10.1097/CM9.0000000000000905 [doi] PST - ppublish SO - Chin Med J (Engl). 2020 Aug 5;133(15):1815-1823. doi: 10.1097/CM9.0000000000000905.