Received:29 November 2019,
Published:30 April 2020
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Lina PAN, Zeyang WANG, Liyuan CHEN, et al. Effect of platelet microvesicles on the function of human microvascular endothelial cells[J]. Journal of Third Military Medical University, 2020, 42(8): 758-764.
Lina PAN, Zeyang WANG, Liyuan CHEN, et al. Effect of platelet microvesicles on the function of human microvascular endothelial cells[J]. Journal of Third Military Medical University, 2020, 42(8): 758-764. DOI: 10.16016/j.1000-5404.201911253.
Objective
2
To investigate the effect of platelet microvesicles (PMVs) on the function of human microvascular endothelial cells (HMECs).
Methods
2
Blood samples from healthy volunteers were collected and anticoagulated with one-tenth volume of citrate solution to isolate platelet-rich plasma (PRP). PMVs released from adenosine diphosphate (ADP)-activated human platelets was collected by centrifugation. HMECs were cultured and grouped as following: Tyrode's buffer as control (Con) group
final concentration of PMVs at 10
20
50 μg/mL respectively
and supernatant (Sup) of PMVs (almost PMVs free) for 24 h. ELISA was used to detect the contents of thromboxane B2 (TXB2) and 6-keto-PGF1α
the stable metabolites of TXA2 and prostaglandin I2 (PGI2) respectively
intercellular adhesion molecule-1 (ICAM-1)
vascular cell adhesion molecule-1 (VCAM-1) and E-selectin in the culture supernatant of HMECs. The expression of endothelin-1 (ET-1) and endothelial nitric oxide synthase (eNOS) phosphorylation in HMECs were determined by Western blotting. Nitric oxide (NO) production in HMECs was measured by fluorescent probe. Endothelial microvesicles (EMVs) secreted by HMECs in the culture supernatant were determined by flow cytometry.
Results
2
Compared with Con group and Sup group
PMVs significantly upregulated the level of TXB2 and decreased that of 6-keto-PGF1α in the supernatant of HMECs in a time- and dose-dependent manner (
P
<
0.05). After incubation with PMVs
the phosphorylated eNOS and the production of NO in HMECs were reduced
while the expression levels of ET-1
ICAM-1
VCAM-1 and E-selectin in the supernatant of HMECs were increased significantly (
P
<
0.05). The concentration of EMVs induced by 50 μg/mL PMVs for 24 h was were increased obviously when compared with con group and Sup group (
P
<
0.05).
Conclusion
2
PMVs may induce the to dysfunction and damage of microvascular endothelial cells. This might be one of the important mechanisms by which platelets are involved in microcirculation dysfunction.
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