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Overexpression of stearoyl-CoA desaturase 1 in bone marrow mesenchymal stem cells enhance the expression of induced endothelial cells.

Lu Y, Zhou Z, Tao J, Dou B, Gao M, Liu Y - Lipids Health Dis (2014)

Bottom Line: Consumption of SFAs impairs arterial endothelial function, while a Mediterranean-type MUFA-diet has a beneficial effect on endothelial function by producing a decrease in levels of vWF, TFPI and PAI-1.High expression of SCD1 is corresponded with low rates of fatty acid oxidation, therefore it might reduce inflammatory responses and be beneficial for the growth of induced endothelial cells.This study suggested that overexpression of SCD1 in BM-MSCs could increase the expression of induced endothelial cells in vitro.

View Article: PubMed Central - HTML - PubMed

Affiliation: Department of Orthopedics, Shanghai First People's Hospital, School of Medicine, Shanghai Jiao Tong University, 100, Haining Road, Shanghai 200080, China. taojie123@sjtu.edu.cn.

ABSTRACT

Background: Bone marrow mesenchymal stem cells (BM-MSCs) are capable of differentiating into endothelial cells in vitro and acquire major characteristics of mature endothelial-like expression of vWF and CD31. SFAs and lipid oxidation products have been linked with postprandial endothelial dysfunction. Consumption of SFAs impairs arterial endothelial function, while a Mediterranean-type MUFA-diet has a beneficial effect on endothelial function by producing a decrease in levels of vWF, TFPI and PAI-1. Stearoyl-CoA desaturase 1 (SCD1), which converts SFA to MUFA, is involved in the cellular biosynthesis of MUFAs from SFA substrates. High expression of SCD1 is corresponded with low rates of fatty acid oxidation, therefore it might reduce inflammatory responses and be beneficial for the growth of induced endothelial cells. Overexpression of SCD1 in BM-MSCs might increase the growth of induced endothelial cells. The goal of this research is to study the relationship between overexpression of SCD1 and the expression of induced endothelial cells in BM-MSCs in vitro.

Methods: The gene SCD1 was integrated into a lentiviral vector, and then 293 T cells were transfected by the connected product to produce a packaged virus. BM-MSCs were infected by the packaged virus. Cell culture and endothelial induction were performed. Fluorescent quantitative PCR of CD31, vWF and VE-cad was performed after 1 week and 2 weeks to test the growth of induced endothelial cells.

Results: The mRNA amount of CD31, vWF and VE-cad of the SCD1 overexpressed group was statistically higher than that of the empty vector (EV) group and that of the normal group after 1 week and 2 weeks, respectively (p < 0.05). Immunocytochemical staining of CD31 or vWF was detected by visualizing red color.

Conclusions: This study suggested that overexpression of SCD1 in BM-MSCs could increase the expression of induced endothelial cells in vitro.

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Vector construction of SCD1 overexpression. (a) An electrophoresis map of the PCR products of SCD1. (b) An electrophoresis map of the PCR products of connected vector pCDH-SCD1. (c) Western blot of SCD1 showed that the concentration of SCD1 in the SCD1 overexpressed group is higher than that in the EV group.
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Figure 3: Vector construction of SCD1 overexpression. (a) An electrophoresis map of the PCR products of SCD1. (b) An electrophoresis map of the PCR products of connected vector pCDH-SCD1. (c) Western blot of SCD1 showed that the concentration of SCD1 in the SCD1 overexpressed group is higher than that in the EV group.

Mentions: RT-PCR showed that the amount of SCD1 mRNA in the SCD1 overexpressed group was 8.86 ± 0.94 and that the amount of the empty vector (EV) group was 1.00 ± 0.09. The expression of SCD1 mRNA of the SCD1 overexpressed group is statistically higher than that of the EV group (p < 0.01) (Figure 2a). Enzyme activity measurement showed that the activity of SCD1 in the SCD1 overexpressed group was 35.87 ± 3.11 and that the activity of the empty vector (EV) group was 23.75 ± 2.16. SCD1 activity of the SCD1 overexpressed group is statistically higher than that of the EV group (p < 0.01) (Figure 2b). A western blot of SCD1 showed that the concentration of SCD1 in the SCD1 overexpressed group was higher than that of the EV group (Figure 3).


Overexpression of stearoyl-CoA desaturase 1 in bone marrow mesenchymal stem cells enhance the expression of induced endothelial cells.

Lu Y, Zhou Z, Tao J, Dou B, Gao M, Liu Y - Lipids Health Dis (2014)

Vector construction of SCD1 overexpression. (a) An electrophoresis map of the PCR products of SCD1. (b) An electrophoresis map of the PCR products of connected vector pCDH-SCD1. (c) Western blot of SCD1 showed that the concentration of SCD1 in the SCD1 overexpressed group is higher than that in the EV group.
© Copyright Policy - open-access
Related In: Results  -  Collection

License 1 - License 2
Show All Figures
getmorefigures.php?uid=PMC3974181&req=5

Figure 3: Vector construction of SCD1 overexpression. (a) An electrophoresis map of the PCR products of SCD1. (b) An electrophoresis map of the PCR products of connected vector pCDH-SCD1. (c) Western blot of SCD1 showed that the concentration of SCD1 in the SCD1 overexpressed group is higher than that in the EV group.
Mentions: RT-PCR showed that the amount of SCD1 mRNA in the SCD1 overexpressed group was 8.86 ± 0.94 and that the amount of the empty vector (EV) group was 1.00 ± 0.09. The expression of SCD1 mRNA of the SCD1 overexpressed group is statistically higher than that of the EV group (p < 0.01) (Figure 2a). Enzyme activity measurement showed that the activity of SCD1 in the SCD1 overexpressed group was 35.87 ± 3.11 and that the activity of the empty vector (EV) group was 23.75 ± 2.16. SCD1 activity of the SCD1 overexpressed group is statistically higher than that of the EV group (p < 0.01) (Figure 2b). A western blot of SCD1 showed that the concentration of SCD1 in the SCD1 overexpressed group was higher than that of the EV group (Figure 3).

Bottom Line: Consumption of SFAs impairs arterial endothelial function, while a Mediterranean-type MUFA-diet has a beneficial effect on endothelial function by producing a decrease in levels of vWF, TFPI and PAI-1.High expression of SCD1 is corresponded with low rates of fatty acid oxidation, therefore it might reduce inflammatory responses and be beneficial for the growth of induced endothelial cells.This study suggested that overexpression of SCD1 in BM-MSCs could increase the expression of induced endothelial cells in vitro.

View Article: PubMed Central - HTML - PubMed

Affiliation: Department of Orthopedics, Shanghai First People's Hospital, School of Medicine, Shanghai Jiao Tong University, 100, Haining Road, Shanghai 200080, China. taojie123@sjtu.edu.cn.

ABSTRACT

Background: Bone marrow mesenchymal stem cells (BM-MSCs) are capable of differentiating into endothelial cells in vitro and acquire major characteristics of mature endothelial-like expression of vWF and CD31. SFAs and lipid oxidation products have been linked with postprandial endothelial dysfunction. Consumption of SFAs impairs arterial endothelial function, while a Mediterranean-type MUFA-diet has a beneficial effect on endothelial function by producing a decrease in levels of vWF, TFPI and PAI-1. Stearoyl-CoA desaturase 1 (SCD1), which converts SFA to MUFA, is involved in the cellular biosynthesis of MUFAs from SFA substrates. High expression of SCD1 is corresponded with low rates of fatty acid oxidation, therefore it might reduce inflammatory responses and be beneficial for the growth of induced endothelial cells. Overexpression of SCD1 in BM-MSCs might increase the growth of induced endothelial cells. The goal of this research is to study the relationship between overexpression of SCD1 and the expression of induced endothelial cells in BM-MSCs in vitro.

Methods: The gene SCD1 was integrated into a lentiviral vector, and then 293 T cells were transfected by the connected product to produce a packaged virus. BM-MSCs were infected by the packaged virus. Cell culture and endothelial induction were performed. Fluorescent quantitative PCR of CD31, vWF and VE-cad was performed after 1 week and 2 weeks to test the growth of induced endothelial cells.

Results: The mRNA amount of CD31, vWF and VE-cad of the SCD1 overexpressed group was statistically higher than that of the empty vector (EV) group and that of the normal group after 1 week and 2 weeks, respectively (p < 0.05). Immunocytochemical staining of CD31 or vWF was detected by visualizing red color.

Conclusions: This study suggested that overexpression of SCD1 in BM-MSCs could increase the expression of induced endothelial cells in vitro.

Show MeSH
Related in: MedlinePlus