PMID- 32029034
OWN - NLM
STAT- MEDLINE
DCOM- 20200217
LR  - 20200217
IS  - 2095-4352 (Print)
VI  - 31
IP  - 12
DP  - 2019 Dec
TI  - [Role of adenosine A2b receptors in pulmonary microvascular endothelial 
      inflammation induced by lipopolysaccharide].
PG  - 1485-1490
LID - 10.3760/cma.j.issn.2095-4352.2019.12.010 [doi]
AB  - OBJECTIVE: To explore the role of the low-affinity A2b adenosine receptors 
      (Adora2b) in pulmonary microvascular endothelial inflammation induced by 
      lipopolysaccharide and its mechanism. METHODS: Rat pulmonary microvascular 
      endothelial cells (PMVECs) were isolated and cultured in vitro. After serum 
      deprivation for 24 hours, cells were pretreated with Adora2b specific agonist 
      BAY60-6583 (0.1, 1, 10 mumol/L) or Adora2b specific antagonist PSB1115 (1 mumol/L) 
      for 1 hour, respectively, and then challenged with LPS (100 mug/L). Cells without 
      treatment were served as the control group, and those treated with LPS, 
      BAY60-6583 or PSB1115 alone were served as single challenge groups. After 
      incubation with specific drugs for 24 hours, the apoptosis of PMVECs was analyzed 
      by flow cytometry using Annexin V/propidium iodide (PI) technique. The levels of 
      early inflammatory factors in cultured medium were measured using enzyme linked 
      immunosorbent assay (ELISA). The mRNA expressions of chemotactic factors and 
      adhesion molecules were determined by real-time quantitative-polymerase chain 
      reaction (RT-qPCR). Polymorph nuclear neutrophils (PMNs) from venous blood of 
      healthy rats were isolated, and PMN migration through PMVECs monolayer under 
      stimulation of drugs was observed in transwell inserts. The monolayer 
      permeability of PMVECs after adhesion of PMNs was determined by fluorescein 
      isothiocyanate (FITC)-albumin assay. Oxidative stress was detected by DCFH-DA 
      assay. RESULTS: Compared with the control group, more cells entered into the 
      apoptosis stage after LPS challenge. Meanwhile, the levels of interleukin-1beta 
      (IL-1beta) and tumor necrosis factor-alpha (TNF-alpha) in cultured medium were significantly 
      increased, as well as the mRNA expressions of chemotactic factors [C-X-C motif 
      chemokine ligand 1 (CXCL-1), CXCL-3 and monocyte chemoattractant protein-1 
      (MCP-1)] and adhesion molecules [E-selectin, intercellular adhesion molecule-1 
      (ICAM-1) and vascular cell adhesion molecule-1 (VCAM-1)]. More PMNs migrated 
      through PMVECs following adhesion and the monolayer permeability of PMVECs was 
      rapidly enhanced. The oxidative stress was upregulated. Compared with LPS group, 
      BAY60-6583 pretreatment could dose-dependently decrease the rate of apoptosis, 
      attenuate trans-endothelial migration of PMNs and decrease the endothelial cell 
      barrier leakage. There were significant differences even after incubation of 0.1 
      mumol/L BAY60-6583 [apoptosis rate: (21.12+/-2.12)% vs. (27.66+/-3.57)%, number of 
      migrated PMNs/HP: 260.60+/-18.24 vs. 290.20+/-16.48, permeability coefficient (Pd, 
      x10(-6) cm/s): 28.28+/-2.04 vs. 32.55+/-2.13, all P < 0.05]. Meanwhile, BAY60-6583 
      pretreatment also downregulated the levels of early proinflammatory factors in a 
      dose-dependent manner as well as the mRNA expressions of chemotactic factors and 
      adhesion molecules. The statistic difference was significant while treated with 1 
      mumol/L BAY60-6583 [IL-1beta (ng/L): 475.75+/-63.15 vs. 755.25+/-67.42, TNF-alpha (ng/L): 
      560.25+/-69.96 vs. 818.75+/-60.92, CXCL-1 mRNA (2(-DeltaDeltaCt)): 3.57+/-0.28 vs. 5.27+/-0.69, 
      CXCL-3 mRNA (2(-DeltaDeltaCt)): 4.56+/-0.48 vs. 7.32+/-0.54, MCP-1 mRNA (2(-DeltaDeltaCt)): 2.21+/-0.31 
      vs. 3.35+/-0.21, E-selectin mRNA (2(-DeltaDeltaCt)): 4.64+/-0.09 vs. 7.28+/-0.73, ICAM-1 mRNA 
      (2(-DeltaDeltaCt)): 4.14+/-0.30 vs. 5.89+/-0.25, VCAM-1 mRNA (2(-DeltaDeltaCt)): 2.23+/-0.19 vs. 
      2.92+/-0.33, all P < 0.05]. Furthermore, pretreatment of 10 mumol/L BAY60-6583 could 
      decrease the oxidative stress [reactive oxygen species (RFU): 629.05+/-33.10 vs. 
      781.45+/-64.59, P < 0.05]. Contrast, PSB1115 pretreatment aggravated apoptosis of 
      PMVECs after LPS incubation [(34.36+/-4.57)% vs. (27.66+/-3.57)%], upregulated 
      expressions of proinflammatory and chemotactic factors as well as adhesion 
      molecules [IL-1beta (ng/L): 889.00+/-63.11 vs. 755.25+/-67.42, TNF-alpha (ng/L): 
      939.00+/-43.44 vs. 818.75+/-60.92, CXCL-1 mRNA (2(-DeltaDeltaCt)): 6.66+/-0.65 vs. 5.27+/-0.69, 
      CXCL-3 mRNA (2(-DeltaDeltaCt)): 10.42+/-0.51 vs. 7.32+/-0.54, MCP-1 mRNA (2(-DeltaDeltaCt)): 
      4.85+/-0.34 vs. 3.35+/-0.21, E-selectin mRNA (2(-DeltaDeltaCt)): 8.42+/-0.47 vs. 7.28+/-0.73, 
      ICAM-1 mRNA (2(-DeltaDeltaCt)): 7.46+/-0.72 vs. 5.89+/-0.25, VCAM-1 mRNA (2(-DeltaDeltaCt)): 
      4.35+/-0.26 vs. 2.92+/-0.33], aggravated trans-endothelial migration of PMNs 
      (cells/HP: 348.40+/-22.68 vs. 290.20+/-16.48), enhanced the leakage of PMVECs 
      monolayer [Pd (x10(-6) cm/s): 39.65+/-2.69 vs. 32.55+/-2.13] and increased oxidative 
      stress in PMVECs [reactive oxygen species (RFU): 847.04+/-29.26 vs. 781.45+/-64.59], 
      with statistically significant difference (all P < 0.05). CONCLUSIONS: Activation 
      of endothelial Adora2b attenuates LPS-induced pulmonary microvascular 
      inflammation by decreasing the release of early inflammatory factors, 
      downregulating expressions of chemotactic factors and adhesion molecules, 
      attenuating trans-endothelial migration of PMNs and oxidative stress in PMVECs, 
      which suggest endothelial Adora2b is apotential anti-inflammatory target in the 
      treatment of LPS-induced acute lung injury.
FAU - Guo, Xiaoxia
AU  - Guo X
AD  - Department of Intensive Care Unit, People's Hospital, Peking University, Beijing 
      100044, China. Corresponding author: An Youzhong, Email: ayzbjicu@163.com.
FAU - An, Youzhong
AU  - An Y
LA  - chi
PT  - Journal Article
PL  - China
TA  - Zhonghua Wei Zhong Bing Ji Jiu Yi Xue
JT  - Zhonghua wei zhong bing ji jiu yi xue
JID - 101604552
RN  - 0 (Lipopolysaccharides)
RN  - 0 (Receptor, Adenosine A2B)
RN  - 0 (Tumor Necrosis Factor-alpha)
SB  - IM
MH  - Animals
MH  - Endothelial Cells
MH  - Inflammation
MH  - Lipopolysaccharides/*metabolism
MH  - *Pneumonia
MH  - Rats
MH  - Receptor, Adenosine A2B/*metabolism
MH  - Tumor Necrosis Factor-alpha
EDAT- 2020/02/08 06:00
MHDA- 2020/02/18 06:00
CRDT- 2020/02/08 06:00
PHST- 2020/02/08 06:00 [entrez]
PHST- 2020/02/08 06:00 [pubmed]
PHST- 2020/02/18 06:00 [medline]
AID - 10.3760/cma.j.issn.2095-4352.2019.12.010 [doi]
PST - ppublish
SO  - Zhonghua Wei Zhong Bing Ji Jiu Yi Xue. 2019 Dec;31(12):1485-1490. doi: 
      10.3760/cma.j.issn.2095-4352.2019.12.010.