RNA interference inhibits Weibel-Palade body release from endothelial cells

doi:10.3969/j.issn.2095-4344.2013.02.021

Abstract

Abstract:

BACKGROUND: RNA interference and silent genes have got more and more attention as they can achieve the clinical effect.
OBJECTIVE: To explore the effect of small hairpin RNA in inhibiting the Weibel-Palade body release in endothelial cell.
METHODS: A small hairpin RNA mediated with an adenovirus vector was designed depending upon the N-terminal functional area of N-ethylmaleimide sensitive factor which act as a key protein in Weibel-Palade body release in endothelial cell. Being screened and identified, the harvested viruses were transfected into human aortic endothelial cell. The adenovirus vector with N-ethylmaleimide sensitive factor small hairpin RNA was regarded as the experimental group, and the pure adenovirus vector was regarded as negative control group, the blank control group without any treatment.
RESULTS AND CONCLUSION: There was significant difference of N-ethylmaleimide sensitive factor mRNA expression in the endothelial cell in three groups after transfected with N-ethylmaleimide sensitive factor small hairpin RNA (P < 0.05); the expression of N-ethylmaleimide sensitive factor mRNA in the experimental group was decreased with time prolonged, and there was significant difference of the N-ethylmaleimide sensitive factor mRNA expression in the experimental group at 24, 48 and 72 hours (P=0.048). The expression of N-ethylmaleimide sensitive factor protein in the experimental group was lower than that in the negative control group and blank control group, and the difference was significant (P < 0.05); there was no significant difference between negative control group and blank control group (P=0.249). The immunofluorescence staining showed that the release of Weibel-Palade body in endothelial cell induced with thrombin was inhibited obviously after transfected with N-ethylmaleimide sensitive factor small hairpin RNA. The expression of N-ethylmaleimide sensitive factor mRNA and protein in endothelial cell transfected with harvested adenoviruses which carried the N-ethylmaleimide sensitive factor small hairpin RNA were significantly decreased, and the release of Weibel-Palade Body induced with thrombin was inhibited dramatically.

Key words: tissue construction, cytological experiments of tissue construction, Weibel-Palade body, RNA interference, N-ethylmaleimide-sensitive factor, endothelial cells, adenovirus, provincial grants-supported paper, tissue construction photographs-containing paper

CLC Number:

R318

Yang Shui-xiang, Wei Xiao-fei, Cui Yu-xia, Zhou Yong. RNA interference inhibits Weibel-Palade body release from endothelial cells[J]. Chinese Journal of Tissue Engineering Research, 2013, 17(2): 301-308.

Figures/Tables 9

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