PMID- 25665603
OWN - NLM
STAT- MEDLINE
DCOM- 20160330
LR  - 20210929
IS  - 2095-4352 (Print)
VI  - 27
IP  - 2
DP  - 2015 Feb
TI  - [Unfractionated heparin inhibits lipopolysaccharide-induced expression of 
      granulocyte colony-stimulating factor in human endothelial cells through 
      Toll-like receptor 4 signaling pathway].
PG  - 81-5
LID - 10.3760/cma.j.issn.2095-4352.2015.02.001 [doi]
AB  - OBJECTIVE: To determine the effect of unfractionated heparin (UFH) on 
      lipopolysaccharide (LPS)-induced expression of granulocyte colony-stimulating 
      factor (G-CSF), and the role of Toll-like receptor 4 (TLR4) signaling pathway in 
      this process. METHODS: Human pulmonary microvascular endothelial cells (HPMECs) 
      were cultured in vitro, and the cells between passages 3 and 5 were used in the 
      experiments. Experiment I: the cells were divided into four groups as follows: 
      control group, LPS stimulation group (LPS 10 mug/mL), LPS + 0.1 U/mL UFH group, 
      and LPS + 1 U/mL UFH group. HPMECs in UFH groups were treated with 0.1 U/mL or 1 
      U/mL UFH 15 minutes before LPS stimulation, and HPMECs in control group were 
      treated with an equal volume of phosphate-buffered saline (PBS) instead. The 
      concentrations of interleukin-6 (IL-6) and G-CSF in cell culture supernatants 
      were determined by enzyme linked immunosorbent assay (ELISA) 24 hours after LPS 
      challenge to detect the effect of UFH on HPMECs. Experiment II: HPMECs were 
      treated with 5 mug/mL of rhodobacter sphaeroides LPS (LPS-RS, antagonist for TLR4) 
      4 hours before the addition of PBS or LPS. The concentrations of IL-6 and G-CSF 
      in cell culture supernatants were determined 24 hours after LPS stimulation to 
      detect the effect of TLR4 on LPS-induced HPMEC injury. Experiment III: HPMECs 
      were divided into four groups as before: control group, LPS stimulation group, 
      LPS + 0.1 U/mL UFH group, LPS + 1 U/mL UFH group. Treatments to cells were the 
      same as experiment I. The protein expression of TLR4 in HPMECs was determined by 
      Western Blot 1 hour after LPS stimulation to detect the effect of UFH on TLR4. 
      RESULTS: (1) Compared with control group, the levels of IL-6 and G-CSF in LPS 
      stimulation group were increased [IL-6 (ng/L): 655.9+/-58.3 vs. 75.5+/-18.2, G-CSF 
      (ng/L): 388.7+/-36.2 vs. 35.3+/-12.6, both P < 0.05]. Compared with those of LPS 
      stimulation group, in LPS + 0.1 U/mL UFH group and LPS + 1 U/mL UFH group, the 
      levels of IL-6 and G-CSF were significantly decreased [IL-6 (ng/L): 518.2+/-64.6, 
      489.1+/-75.6 vs. 655.9+/-58.3, G-CSF (ng/L): 298.8+/-41.0, 273.4+/-33.2 vs. 388.7+/-36.2, 
      all P < 0.05]. The results indicated that 1 U/mL UFH had better results, though 
      there was no statistical significance between the results of two UFH groups. (2) 
      LPS-induced up-regulation of IL-6 and G-CSF levels was prevented by LPS-RS [IL-6 
      (ng/L): 139.1+/-37.6 vs. 655.9+/-58.3, G-CSF (ng/L): 73.7+/-19.7 vs. 388.7+/-36.2, both P 
      < 0.05]. LPS-RS alone had no effect on cytokines [IL-6 (ng/L): 118.2+/-42.1 vs. 
      75.5+/-18.2, G-CSF (ng/L): 48.4+/-26.8 vs. 35.3+/-12.6, both P > 0.05]. (3) Compared 
      with control group, the protein expression of TLR4 (grey value) in LPS 
      stimulation group was significantly upregulated after 1 hour (0.87+/-0.23 vs. 
      0.36+/-0.12, P < 0.05). UFH with 0.1 U/mL and 1 U/mL lowered TLR-4 protein 
      expression induced by LPS (0.68+/-0.18, 0.62+/-0.26 vs. 0.87+/-0.23, both P < 0.05). 
      CONCLUSIONS: The expressions of IL-6 and G-CSF were increased obviously in LPS 
      treated HPMECs. UFH might take its therapeutic effect through TLR4-dependent 
      pathway.
FAU - Li, Xu
AU  - Li X
AD  - Department of Critical Care Medicine, the First Affiliated Hospital of China 
      Medical University, Shenyang 110001, Liaoning, China. Corresponding author: Ma 
      Xiaochun, Email: xcma2972@sina.com.
FAU - Liu, Yina
AU  - Liu Y
FAU - Ma, Xiaochun
AU  - Ma X
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 (Cytokines)
RN  - 0 (Interleukin-6)
RN  - 0 (Lipopolysaccharides)
RN  - 0 (TLR4 protein, human)
RN  - 0 (Toll-Like Receptor 4)
RN  - 143011-72-7 (Granulocyte Colony-Stimulating Factor)
RN  - 9005-49-6 (Heparin)
SB  - IM
MH  - Cells, Cultured
MH  - Cytokines
MH  - Endothelial Cells/*drug effects
MH  - Granulocyte Colony-Stimulating Factor/*drug effects
MH  - Heparin/*pharmacology
MH  - Humans
MH  - Interleukin-6
MH  - Lipopolysaccharides
MH  - Lung
MH  - *Signal Transduction
MH  - Toll-Like Receptor 4/*metabolism
EDAT- 2015/02/11 06:00
MHDA- 2016/03/31 06:00
CRDT- 2015/02/11 06:00
PHST- 2015/02/11 06:00 [entrez]
PHST- 2015/02/11 06:00 [pubmed]
PHST- 2016/03/31 06:00 [medline]
AID - 10.3760/cma.j.issn.2095-4352.2015.02.001 [doi]
PST - ppublish
SO  - Zhonghua Wei Zhong Bing Ji Jiu Yi Xue. 2015 Feb;27(2):81-5. doi: 
      10.3760/cma.j.issn.2095-4352.2015.02.001.