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N-ethylmaleimide‑sensitive factor siRNA inhibits the release of Weibel-Palade bodies in endothelial cells.

Zhou Y, Yang SX, Yue YN, Wei XF, Liu Y - Mol Med Rep (2016)

Bottom Line: In addition, the mRNA expression of NSF was gradually decreased as duration increased; there were marked differences between the 24, 48 and 72 h groups (P<0.05).The protein expression of NSF was significantly decreased in the experimental group, compared with the negative control group (P=0.004) and blank control group (P=0.031), however, no difference was observed between the negative control and blank control groups (P=0.249).These results suggested that NSF-siRNA may be valuable for preventing and treating atherosclerosis and acute coronary syndrome.

View Article: PubMed Central - PubMed

Affiliation: Department of Cardiology, Beijing Shijitan Hospital, Capital Medical University, Beijing 100038, P.R. China.

ABSTRACT
The aim of the present study was to examine the effect of small interfering RNA (siRNA) methods on the expression of N‑ethylmaleimide sensitive factor (NSF) and Weibel‑Palade body (WPB) release in endothelial cells. A small hairpin RNA (shRNA), mediated with an adenovirus vector, was designed to target the N‑terminal functional area of NSF. Subsequently, viruses were transfected into human aortic endothelial cells. The mRNA and protein expression levels of NSF were detected using reverse transcription‑quantitative polymerase chain reaction and Western blot analyses, respectively, and the release of WPBs in the endothelial cells was examined using immunofluorescence. The mRNA expression of NSF in the endothelial cells, which were transfected with the adenoviruses carrying the NSF‑shRNA was significantly decreased, compared with the negative control group (P=0.035) and blank control group (P=0.02). In addition, the mRNA expression of NSF was gradually decreased as duration increased; there were marked differences between the 24, 48 and 72 h groups (P<0.05). The protein expression of NSF was significantly decreased in the experimental group, compared with the negative control group (P=0.004) and blank control group (P=0.031), however, no difference was observed between the negative control and blank control groups (P=0.249). The immunofluorescence staining showed that the release of WPBs in the endothelial cells induced with thrombin was inhibited markedly following transfection with the virus carrying the NSF‑shRNA. Therefore NSF‑siRNA inhibited the mRNA and protein expression levels of NSF, and inhibited the release of WPBs in endothelial cells induced with thrombin. These results suggested that NSF-siRNA may be valuable for preventing and treating atherosclerosis and acute coronary syndrome.

No MeSH data available.


Related in: MedlinePlus

shRNA represses the release of WPB in human aortic endothelial cells. (A) Control group; (B) thrombin-induced WPB release; (C) shRNA+thrombin. shRNA repressed thrombin-induced WPB release; (D) dsDNA+thrombin, dsDNA did not repress thrombin-induced WPB release. Magnification, ×60. WPB, Weibel-Palade body; shRNA, short hairpin RNA; ds, double stranded RNA.
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f4-mmr-14-02-1061: shRNA represses the release of WPB in human aortic endothelial cells. (A) Control group; (B) thrombin-induced WPB release; (C) shRNA+thrombin. shRNA repressed thrombin-induced WPB release; (D) dsDNA+thrombin, dsDNA did not repress thrombin-induced WPB release. Magnification, ×60. WPB, Weibel-Palade body; shRNA, short hairpin RNA; ds, double stranded RNA.

Mentions: The immunofluorescence staining showed that the release of WPBs in the thrombin-induced endothelial cells was markedly inhibited following transduction with the virus carrying NSF-shRNA, but was not affected in the negative or blank control groups (Fig. 4).


N-ethylmaleimide‑sensitive factor siRNA inhibits the release of Weibel-Palade bodies in endothelial cells.

Zhou Y, Yang SX, Yue YN, Wei XF, Liu Y - Mol Med Rep (2016)

shRNA represses the release of WPB in human aortic endothelial cells. (A) Control group; (B) thrombin-induced WPB release; (C) shRNA+thrombin. shRNA repressed thrombin-induced WPB release; (D) dsDNA+thrombin, dsDNA did not repress thrombin-induced WPB release. Magnification, ×60. WPB, Weibel-Palade body; shRNA, short hairpin RNA; ds, double stranded RNA.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4940057&req=5

f4-mmr-14-02-1061: shRNA represses the release of WPB in human aortic endothelial cells. (A) Control group; (B) thrombin-induced WPB release; (C) shRNA+thrombin. shRNA repressed thrombin-induced WPB release; (D) dsDNA+thrombin, dsDNA did not repress thrombin-induced WPB release. Magnification, ×60. WPB, Weibel-Palade body; shRNA, short hairpin RNA; ds, double stranded RNA.
Mentions: The immunofluorescence staining showed that the release of WPBs in the thrombin-induced endothelial cells was markedly inhibited following transduction with the virus carrying NSF-shRNA, but was not affected in the negative or blank control groups (Fig. 4).

Bottom Line: In addition, the mRNA expression of NSF was gradually decreased as duration increased; there were marked differences between the 24, 48 and 72 h groups (P<0.05).The protein expression of NSF was significantly decreased in the experimental group, compared with the negative control group (P=0.004) and blank control group (P=0.031), however, no difference was observed between the negative control and blank control groups (P=0.249).These results suggested that NSF-siRNA may be valuable for preventing and treating atherosclerosis and acute coronary syndrome.

View Article: PubMed Central - PubMed

Affiliation: Department of Cardiology, Beijing Shijitan Hospital, Capital Medical University, Beijing 100038, P.R. China.

ABSTRACT
The aim of the present study was to examine the effect of small interfering RNA (siRNA) methods on the expression of N‑ethylmaleimide sensitive factor (NSF) and Weibel‑Palade body (WPB) release in endothelial cells. A small hairpin RNA (shRNA), mediated with an adenovirus vector, was designed to target the N‑terminal functional area of NSF. Subsequently, viruses were transfected into human aortic endothelial cells. The mRNA and protein expression levels of NSF were detected using reverse transcription‑quantitative polymerase chain reaction and Western blot analyses, respectively, and the release of WPBs in the endothelial cells was examined using immunofluorescence. The mRNA expression of NSF in the endothelial cells, which were transfected with the adenoviruses carrying the NSF‑shRNA was significantly decreased, compared with the negative control group (P=0.035) and blank control group (P=0.02). In addition, the mRNA expression of NSF was gradually decreased as duration increased; there were marked differences between the 24, 48 and 72 h groups (P<0.05). The protein expression of NSF was significantly decreased in the experimental group, compared with the negative control group (P=0.004) and blank control group (P=0.031), however, no difference was observed between the negative control and blank control groups (P=0.249). The immunofluorescence staining showed that the release of WPBs in the endothelial cells induced with thrombin was inhibited markedly following transfection with the virus carrying the NSF‑shRNA. Therefore NSF‑siRNA inhibited the mRNA and protein expression levels of NSF, and inhibited the release of WPBs in endothelial cells induced with thrombin. These results suggested that NSF-siRNA may be valuable for preventing and treating atherosclerosis and acute coronary syndrome.

No MeSH data available.


Related in: MedlinePlus