PMID- 26911942
OWN - NLM
STAT- MEDLINE
DCOM- 20161020
LR  - 20190816
IS  - 2095-4352 (Print)
VI  - 28
IP  - 2
DP  - 2016 Feb
TI  - [Unfractionated heparin inhibits lipopolysaccharide-induced expression of 
      chemokines in human endothelial cells through nuclear factor-KappaB signaling 
      pathway].
PG  - 117-21
LID - 10.3760/cma.j.issn.2095-4352.2016.02.007 [doi]
AB  - OBJECTIVE: To determine the effect of unfractionated heparin (UFH) on 
      lipopolysaccharide (LPS)-induced expression of chemokines and nuclear factor-KappaB 
      (NF-KappaB) signaling pathway. METHODS: Human pulmonary microvascular endothelial 
      cells (HPMECs) were cultured in vitro, and the cells between passages 3 and 5 
      were used in the experiments. The cells were divided into control group, LPS 
      challenge group, 1 kU/L or 10 kU/L UFH+LPS group, and NF-KappaB inhibitor 
      N-tosyl-L-lysyl chloromethyl-ketone (TLCK) group (TLCK+LPS group). HPMECs in LPS 
      challenge group were treated with 10 mg/L LPS. UFH pretreatment with different 
      dosages groups were treated with 1 kU/L or 10 kU/L UFH 15 minutes before LPS 
      challenge. Cells in the TLCK+LPS group were treated with 10 mumol/L of TLCK 30 
      minutes before the addition of LPS, and HPMECs in control group were treated with 
      an equal volume of phosphate-buffered saline (PBS) instead. The cells were 
      harvested 1 hour after LPS challenge, and the nuclear translocation of NF-KappaB was 
      determined by immunofluorescence assay to detect the effect of UFH on NF-KappaB 
      activation. The levels of interleukin-8 (IL-8) and monocyte chemoattractant 
      protein-1 (MCP-1) in cell culture supernatants were determined by enzyme linked 
      immunosorbent assay (ELISA) 3 hours and 6 hours after LPS challenge to detect the 
      effect of UFH on LPS induced expression of chemokines and its mechanism of effect 
      on NF-KappaB signaling pathway in HPMECs. RESULTS: (1) In the control group, NF-KappaB 
      was mostly located in the cytosol as shown by immunofluorescence. Treatment of 
      HPMECs with LPS significantly increased the translocation of NF-KappaB from the 
      cytosol to nucleus. UFH suppressed LPS-induced NF-KappaB activation both in 1 kU/L 
      and 10 kU/L dosages, and 10 kU/L UFH gave even better results. (2) Compared with 
      control group, the levels of IL-8 and MCP-1 in the supernatants in LPS challenge 
      group were significantly increased at 3 hours and 6 hours after LPS challenge 
      [IL-8 (ng/L): 387.1+/-26.4 vs. 23.8+/-8.1 at 3 hours, 645.5+/-69.6 vs. 125.7+/-18.7 at 6 
      hours; MCP-1 (ng/L): 3 654.9+/-467.9 vs. 721.6+/-61.3 at 3 hours, 8 178.5+/-792.6 vs. 1 
      324.7+/-148.7 at 6 hours, all P < 0.05]. Compared with that of LPS challenge group, 
      in 1 kU/L and 10 kU/L UFH pretreatment groups, the levels of IL-8 and MCP-1 were 
      significantly decreased [IL-8 (ng/L): 315.3+/-24.8, 275.8+/-31.1 vs. 387.1+/-26.4 at 3 
      hours, 557.8+/-43.3, 496.9+/-38.7 vs. 645.5+/-69.6 at 6 hours; MCP-1 (ng/L): 2 
      924.1+/-267.9, 2 668.3+/-522.6 vs. 3 654.9+/-467.9 at 3 hours, 7 121.7+/-557.2, 6 
      563.9+/-576.4 vs. 8 178.5+/-792.6 at 6 hours, all P < 0.05]. The results indicated 
      that 10 kU/L UFH yielded better results. However, inhibition study using the 
      known NF-KappaB inhibitor TLCK could decrease LPS-induced increase in IL-8 and MCP-1 
      levels [IL-8 (ng/L): 162.4+/-21.3 vs. 387.1+/-26.4 at 3 hours, 274.1+/-22.6 vs. 
      645.5+/-69.6 at 6 hours; MCP-1 (ng/L): 1 478.2+/-138.5 vs. 3 654.9+/-467.9 at 3 hours; 
      3 667.6+/-259.4 vs. 8 178.5+/-792.6 at 6 hours, all P < 0.05]. CONCLUSIONS: The 
      levels of IL-8 and MCP-1 were increased obviously in LPS treated HPMECs. UFH 
      might suppress LPS-activated NF-KappaB signaling pathway, contributing to the 
      inhibitory effects of chemokines in HPMECs.
FAU - Li, Xu
AU  - Li X
AD  - Department of Critical Care Medicine, the First Affiliated Hospital of China 
      Medical University, Shenyang 110001, Liaoning, China (Li X, Chen TL, Tang J, Ma 
      XC); Department of Critical Care Medicine, General Hospital of Panjin Oilfield, 
      Panjin 124010, Liaoning, China (Ma YQ). Corresponding author: Ma Xiaochun, Email: 
      xcma2972@sina.com.
FAU - Ma, Yanquan
AU  - Ma Y
FAU - Chen, Tianlu
AU  - Chen T
FAU - Tang, Jie
AU  - Tang J
FAU - Ma, Xiaochun
AU  - Ma X
LA  - chi
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
PL  - China
TA  - Zhonghua Wei Zhong Bing Ji Jiu Yi Xue
JT  - Zhonghua wei zhong bing ji jiu yi xue
JID - 101604552
RN  - 0 (CCL2 protein, human)
RN  - 0 (CXCL8 protein, human)
RN  - 0 (Chemokine CCL2)
RN  - 0 (Interleukin-8)
RN  - 0 (Lipopolysaccharides)
RN  - 0 (NF-kappa B)
RN  - 9005-49-6 (Heparin)
SB  - IM
MH  - Cells, Cultured
MH  - Chemokine CCL2/*metabolism
MH  - Endothelial Cells/cytology/*drug effects
MH  - Heparin/*pharmacology
MH  - Humans
MH  - Interleukin-8/*metabolism
MH  - Lipopolysaccharides
MH  - Lung/cytology
MH  - NF-kappa B/*metabolism
MH  - *Signal Transduction
EDAT- 2016/02/26 06:00
MHDA- 2016/10/21 06:00
CRDT- 2016/02/26 06:00
PHST- 2016/02/26 06:00 [entrez]
PHST- 2016/02/26 06:00 [pubmed]
PHST- 2016/10/21 06:00 [medline]
AID - 10.3760/cma.j.issn.2095-4352.2016.02.007 [doi]
PST - ppublish
SO  - Zhonghua Wei Zhong Bing Ji Jiu Yi Xue. 2016 Feb;28(2):117-21. doi: 
      10.3760/cma.j.issn.2095-4352.2016.02.007.