Changes of endothelial microparticles induced by lipopolysaccharide

doi:10.3877/cma.j.issn.1673-9450.2017.02.006

Abstract

Abstract:

Objective

To investigate the significance of changes of endothelial microparticle(EMP) induced by lipopolysaccharide.

Methods

Human umbilical vein endothelial cell (HUVEC) was cultured. The cells were divided into control group (without lipopolysaccharide) and lipopolysaccharide group (0.1, 1.0, 10.0, 20.0 μg/mL). The cell culture supernatant were collected after 24 h incubation with or without lipopolysaccharide. The cells were digested and suspended. EMP was collected by series of centrifugation and incubated with CFS anti-CD31, PE anti-CD51, FITC anti-CD54, PE anti-CD144, APC anti-CD62E. Flow cytometry was performed to analyze by detecting the expression of the fluorescenct labeling EMP and the measurement of apoptosis of endotheliums. Data were processed with analysis of variance, t and Dunnett-t test.

Results

The counts of CD31⁺EMP in control group and lipopolysaccharide group (0.1, 1.0, 10.0, 20.0 μg/mL) were respectively (1.23±0.43)×10^6, (1.92±0.64)×10^6, (2.55±0.78)×10^6, (3.96±0.78)×10^6, (6.96±1.80)×10^6, there were significant differences (F=5.83, P<0.05), and the ratios of CD31⁺EMP were respectively (4.57±1.33)%, (5.92±1.24)%, (6.93±1.61)%, (7.44±1.09)%, (13.44±3.39)%, there were significant differences (F=3.43, P<0.05). Compared with that of control group, the counts of CD31⁺EMP were obviously increased in 10.0, 20.0 μg/mL lipopolysaccharide groups (with t value respectively -3.06, -3.61, P values were less than 0.05). In 20.0 μg/mL lipopolysaccharide group, the counts of CD31⁺EMP were higher than that in 0.1, 1.0 μg/mL lipopolysaccharide groups (P<0.05), and the ratios of CD31⁺EMP were higher than that in control group (t=-2.74, P<0.05) and in 0.1 μg/mL lipopolysaccharide group (P<0.05). The counts of CD54⁺EMP in control group and lipopolysaccharide group (0.1, 1.0, 10.0, 20.0 μg/mL) were respectively (4.70±1.01)×10^6, (9.20±3.34)×10^6, (8.83±1.70)×10^6, (17.18±3.78)×10^6, (17.73±4.98)×10^6, there were significant differences (F=3.35, P<0.05). Compared with that of control group, the counts of CD54⁺EMP were obviously increased in 10.0, 20.0 μg/mL lipopolysaccharide groups (with t value respectively -3.19, -3.00, P values were less than 0.05). The counts of CD144⁺EMP in control group and lipopolysaccharide group (0.1, 1.0, 10.0, 20.0 μg/mL) were respectively (3.93±1.22)×10^6, (6.80±1.36)×10^6, (8.03±2.53)×10^6, (17.22±4.52)×10^6, (11.05±3.80)×10^6, there were significant differences (F=3.17, P<0.05). The counts of CD144⁺EMP were higher in 10.0 μg/mL lipopolysaccharide group than that in control group and that in 0.1, 1.0 μg/mL lipopolysaccharide groups (P values were less than 0.05). In 20.0 μg/mL lipopolysaccharide group, the counts of CD62E⁺EMP were (2.95±0.26)×10^6, which were higher than that in control group whose counts were (1.26±0.26)×10⁶(t=-4.45, P<0.05). Comparing the apoptosis rate of endothelial cell in each group, there were no significant differences (F=0.17, P>0.05).

Conclusion

The increase of EMP levels suggests that the specific bioactivity of each antigen-positive EMP may participate in regulating endothelial function induced by lipopolysaccharide, and they may be one of the mechanisms of endothelial dysfunction in sepsis.

Key words: Lipopolysaccharides, Endothelial cells, Umbilical veins, Microparticles

Jiajun Tang, Jiping Xu, Jiexin Zheng, Qin Zhang. Changes of endothelial microparticles induced by lipopolysaccharide[J]. Chinese Journal of Injury Repair and Wound Healing(Electronic Edition), 2017, 12(02): 103-108.

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Figures/Tables 6

References 22

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组别	CD31^＋EMP(×10⁶)	CD31^＋EMP(%)
对照组	1.23±0.43	4.57±1.33
0.1 μg/mL脂多糖组	1.92±0.64^ae	5.92±1.24^ce
1.0 μg/mL脂多糖组	2.55±0.78^ae	6.93±1.61^c
10.0 μg/mL脂多糖组	3.96±0.78^b	7.44±1.09^c
20.0 μg/mL脂多糖组	6.96±1.80^b	13.44±3.39^d
F值	5.83	3.43
P值	<0.05	<0.05

组别	CD31^＋EMP(×10⁶)	CD31^＋EMP(%)
对照组	1.23±0.43	4.57±1.33
0.1 μg/mL脂多糖组	1.92±0.64^ae	5.92±1.24^ce
1.0 μg/mL脂多糖组	2.55±0.78^ae	6.93±1.61^c
10.0 μg/mL脂多糖组	3.96±0.78^b	7.44±1.09^c
20.0 μg/mL脂多糖组	6.96±1.80^b	13.44±3.39^d
F值	5.83	3.43
P值	<0.05	<0.05

组别	CD54^＋EMP(×10⁶)	CD54^＋EMP(%)
对照组	4.70±1.01	17.92±3.29
0.1 μg/mL脂多糖组	9.20±3.34^a	27.63±6.21^c
1.0 μg/mL脂多糖组	8.83±1.70a	24.93±3.04^c
10.0 μg/mL脂多糖组	17.18±3.78^b	32.15±5.53^c
20.0 μg/mL脂多糖组	17.73±4.98^b	33.77±8.65^c
F值	3.35	1.37
P值	<0.05	>0.05

组别	CD54^＋EMP(×10⁶)	CD54^＋EMP(%)
对照组	4.70±1.01	17.92±3.29
0.1 μg/mL脂多糖组	9.20±3.34^a	27.63±6.21^c
1.0 μg/mL脂多糖组	8.83±1.70a	24.93±3.04^c
10.0 μg/mL脂多糖组	17.18±3.78^b	32.15±5.53^c
20.0 μg/mL脂多糖组	17.73±4.98^b	33.77±8.65^c
F值	3.35	1.37
P值	<0.05	>0.05

组别	CD144^＋EMP(×10⁶)	CD144^＋EMP(%)
对照组	3.93±1.22	16.00±6.11
0.1 μg/mL脂多糖组	6.80±1.36^ad	22.77±5.06^c
1.0 μg/mL脂多糖组	8.03±2.53^ad	23.81±8.24^c
10.0 μg/mL脂多糖组	17.22±4.52^b	32.08±7.62^c
20.0 μg/mL脂多糖组	11.05±3.80^a	21.58±7.82^c
F值	3.17	0.71
P值	<0.05	>0.05

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