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Activated protein C induces endoplasmic reticulum stress and attenuates lipopolysaccharide-induced apoptosis mediated by glycogen synthase kinase-3β.

Luo L, Lv T, Wang Q, Zhang T, Gu X, Xu F, Song Y - Mediators Inflamm. (2012)

Bottom Line: Calcium inhibition did not alter the antiapoptotic effect of activated protein C.The antiapoptotic efficiency of activated protein C was reduced in human umbilical vein endothelial cells following treatment with glycogen synthase kinase-3β-siRNA.In summary, activated protein C induced endoplasmic reticulum stress and attenuated lipopolysaccharide-induced human umbilical vein endothelial cell apoptosis mediated by glycogen synthase kinase-3β.

View Article: PubMed Central - PubMed

Affiliation: Department of Respiratory Medicine, Jinling Hospital, Nanjing University School of Medicine, Nanjing 210002, China.

ABSTRACT
This study investigated the relationship between antiapoptotic activities induced by activated protein C and endoplasmic reticulum stress. In this study, it was observed that activated protein C elicited a rise in glucose-regulated protein 78 and glycogen synthase kinase-3β and inhibited apoptosis in human umbilical vein endothelial cells induced by lipopolysaccharide. Calcium inhibition did not alter the antiapoptotic effect of activated protein C. The antiapoptotic efficiency of activated protein C was reduced in human umbilical vein endothelial cells following treatment with glycogen synthase kinase-3β-siRNA. In summary, activated protein C induced endoplasmic reticulum stress and attenuated lipopolysaccharide-induced human umbilical vein endothelial cell apoptosis mediated by glycogen synthase kinase-3β.

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Upregulation of GSK-3β expression following APC treatment and decreased antiapoptotic activity following GSK-3β-siRNA transfection of HUVECs. HUVECs exposed to 10 ng/mL LPS for 24 h, were treated with 150 nM APC for 0, 6, 12, and 24 h. Upregulated GSK-3β expression was detected, peaking at 6 h after APC treatment (a). Transfection of HUVECs with GSK-3β-siRNA decreased the abundance of GSK-3β protein significantly (b). Compared with normal HUVECs treated with 150 nM APC, a significant increase of the percentage of apoptosis cells was found in HUVECs pretreated with GSK-3β-siRNA (GSK-3β versus GSK-3β-siRNA, P < 0.05), indicating that the antiapoptotic activity of APC was inhibited by GSK-3β-siRNA (c).
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fig4: Upregulation of GSK-3β expression following APC treatment and decreased antiapoptotic activity following GSK-3β-siRNA transfection of HUVECs. HUVECs exposed to 10 ng/mL LPS for 24 h, were treated with 150 nM APC for 0, 6, 12, and 24 h. Upregulated GSK-3β expression was detected, peaking at 6 h after APC treatment (a). Transfection of HUVECs with GSK-3β-siRNA decreased the abundance of GSK-3β protein significantly (b). Compared with normal HUVECs treated with 150 nM APC, a significant increase of the percentage of apoptosis cells was found in HUVECs pretreated with GSK-3β-siRNA (GSK-3β versus GSK-3β-siRNA, P < 0.05), indicating that the antiapoptotic activity of APC was inhibited by GSK-3β-siRNA (c).

Mentions: APC treatment elicited a rise in GSK-3β expression, peaking after 6 h APC treatment. The level of GSK-3β generally decreased with prolonged APC treatment (Figure 4(a)). To further substantiate the role of GSK-3β, GSK-3β-siRNA was used to inhibit the expression of GSK-3β. HUVECs transfected by GSK-3β-siRNA showed a substantial reduction in GSK-3β mRNA compared to the control by RT-PCR analysis (GSK-3β-siRNA versus control, P < 0.05). Transfection of HUVEC with GSK-3β-siRNA decreased the abundance of GSK-3β protein significantly (Figure 4(b)). Compared with normal HUVECs treated with 150 nM APC, a significant increase in the number apoptotic cells was detected in HUVECs pretreated with GSK-3β-siRNA (Figure 4(c)), thus indicating that antiapoptotic activity of APC was partially inhibited by GSK-3β-siRNA.


Activated protein C induces endoplasmic reticulum stress and attenuates lipopolysaccharide-induced apoptosis mediated by glycogen synthase kinase-3β.

Luo L, Lv T, Wang Q, Zhang T, Gu X, Xu F, Song Y - Mediators Inflamm. (2012)

Upregulation of GSK-3β expression following APC treatment and decreased antiapoptotic activity following GSK-3β-siRNA transfection of HUVECs. HUVECs exposed to 10 ng/mL LPS for 24 h, were treated with 150 nM APC for 0, 6, 12, and 24 h. Upregulated GSK-3β expression was detected, peaking at 6 h after APC treatment (a). Transfection of HUVECs with GSK-3β-siRNA decreased the abundance of GSK-3β protein significantly (b). Compared with normal HUVECs treated with 150 nM APC, a significant increase of the percentage of apoptosis cells was found in HUVECs pretreated with GSK-3β-siRNA (GSK-3β versus GSK-3β-siRNA, P < 0.05), indicating that the antiapoptotic activity of APC was inhibited by GSK-3β-siRNA (c).
© Copyright Policy - open-access
Related In: Results  -  Collection

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getmorefigures.php?uid=PMC3368528&req=5

fig4: Upregulation of GSK-3β expression following APC treatment and decreased antiapoptotic activity following GSK-3β-siRNA transfection of HUVECs. HUVECs exposed to 10 ng/mL LPS for 24 h, were treated with 150 nM APC for 0, 6, 12, and 24 h. Upregulated GSK-3β expression was detected, peaking at 6 h after APC treatment (a). Transfection of HUVECs with GSK-3β-siRNA decreased the abundance of GSK-3β protein significantly (b). Compared with normal HUVECs treated with 150 nM APC, a significant increase of the percentage of apoptosis cells was found in HUVECs pretreated with GSK-3β-siRNA (GSK-3β versus GSK-3β-siRNA, P < 0.05), indicating that the antiapoptotic activity of APC was inhibited by GSK-3β-siRNA (c).
Mentions: APC treatment elicited a rise in GSK-3β expression, peaking after 6 h APC treatment. The level of GSK-3β generally decreased with prolonged APC treatment (Figure 4(a)). To further substantiate the role of GSK-3β, GSK-3β-siRNA was used to inhibit the expression of GSK-3β. HUVECs transfected by GSK-3β-siRNA showed a substantial reduction in GSK-3β mRNA compared to the control by RT-PCR analysis (GSK-3β-siRNA versus control, P < 0.05). Transfection of HUVEC with GSK-3β-siRNA decreased the abundance of GSK-3β protein significantly (Figure 4(b)). Compared with normal HUVECs treated with 150 nM APC, a significant increase in the number apoptotic cells was detected in HUVECs pretreated with GSK-3β-siRNA (Figure 4(c)), thus indicating that antiapoptotic activity of APC was partially inhibited by GSK-3β-siRNA.

Bottom Line: Calcium inhibition did not alter the antiapoptotic effect of activated protein C.The antiapoptotic efficiency of activated protein C was reduced in human umbilical vein endothelial cells following treatment with glycogen synthase kinase-3β-siRNA.In summary, activated protein C induced endoplasmic reticulum stress and attenuated lipopolysaccharide-induced human umbilical vein endothelial cell apoptosis mediated by glycogen synthase kinase-3β.

View Article: PubMed Central - PubMed

Affiliation: Department of Respiratory Medicine, Jinling Hospital, Nanjing University School of Medicine, Nanjing 210002, China.

ABSTRACT
This study investigated the relationship between antiapoptotic activities induced by activated protein C and endoplasmic reticulum stress. In this study, it was observed that activated protein C elicited a rise in glucose-regulated protein 78 and glycogen synthase kinase-3β and inhibited apoptosis in human umbilical vein endothelial cells induced by lipopolysaccharide. Calcium inhibition did not alter the antiapoptotic effect of activated protein C. The antiapoptotic efficiency of activated protein C was reduced in human umbilical vein endothelial cells following treatment with glycogen synthase kinase-3β-siRNA. In summary, activated protein C induced endoplasmic reticulum stress and attenuated lipopolysaccharide-induced human umbilical vein endothelial cell apoptosis mediated by glycogen synthase kinase-3β.

Show MeSH