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VEGF is a mediator of the renoprotective effects of multipotent marrow stromal cells in acute kidney injury.

Tögel F, Zhang P, Hu Z, Westenfelder C - J. Cell. Mol. Med. (2008)

Bottom Line: Knockdown of vascular enthothelial growth factor (VEGF) by siRNA reduced effectiveness of MSCs in the treatment of ischemic AKI in a rat model.Animals treated with MSCs had increased renal microvessel density compared to VEGF knockdown MSC-treated and vehicle-treated animals.These results show that VEGF is an important mediator of the early and late phase of renoprotective action after AKI in the context of stem cell treatment.

View Article: PubMed Central - PubMed

Affiliation: University of Utah, Division of Nephrology, Department of Medicine, Salt Lake City, UT 84148, USA.

ABSTRACT
Adult stem cell treatment of complex disorders is a promising therapeutic approach and multipotent marrow stromal cells (MSCs) have been shown to be effective in various animal models of diseases. Acute kidney injury (AKI) is a common and serious problem in hospitalized patients and bone marrow derived multipotent MSCs have been shown to be effective in different models of AKI. The mechanism of action of MSCs is complex but involves paracrine actions including growth factor secretion. Knockdown of vascular enthothelial growth factor (VEGF) by siRNA reduced effectiveness of MSCs in the treatment of ischemic AKI in a rat model. Animals treated with MSCs had increased renal microvessel density compared to VEGF knockdown MSC-treated and vehicle-treated animals. These results show that VEGF is an important mediator of the early and late phase of renoprotective action after AKI in the context of stem cell treatment.

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Related in: MedlinePlus

Knockdown efficiency of VEGF on the mRNA and protein level. Left panel: Absolute gene regulation determined by real time quantitative RT‐PCR. Right panel: VEGF protein concentrations determined by ELISA in tissue culture supernatant of cells treated with VEGF siRNA after 24 and 48 hrs after the end of the siRNA incubation period (24 hrs). VEGF knockdown on mRNA and protein level was highly significant (P < 0.01, t‐test).
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f1: Knockdown efficiency of VEGF on the mRNA and protein level. Left panel: Absolute gene regulation determined by real time quantitative RT‐PCR. Right panel: VEGF protein concentrations determined by ELISA in tissue culture supernatant of cells treated with VEGF siRNA after 24 and 48 hrs after the end of the siRNA incubation period (24 hrs). VEGF knockdown on mRNA and protein level was highly significant (P < 0.01, t‐test).

Mentions: Adherent MSCs in culture flasks were treated for 24 hrs with VEGF siRNA and NeoFX transfection agent in regular growth medium in order to knockdown VEGF expression. Knockdown efficiency was determined at RNA and protein levels 24 hrs and 48 hrs after the end of the transfection period. Efficiency of the knockdown approach was tested before each experiment and adjusted if necessary (combination of siRNAs or different concentrations). Initially, combination of three VEGF siRNA yielded a greater than 80% knockdown of VEGF at the RNA and protein levels. In later experiments, 10 nm of siRNA ID #192613 was used and yielded a greater than 60% knockdown at the protein level. Results are shown in Fig. 1. Knockdown was verified at the mRNA as well as the protein level.


VEGF is a mediator of the renoprotective effects of multipotent marrow stromal cells in acute kidney injury.

Tögel F, Zhang P, Hu Z, Westenfelder C - J. Cell. Mol. Med. (2008)

Knockdown efficiency of VEGF on the mRNA and protein level. Left panel: Absolute gene regulation determined by real time quantitative RT‐PCR. Right panel: VEGF protein concentrations determined by ELISA in tissue culture supernatant of cells treated with VEGF siRNA after 24 and 48 hrs after the end of the siRNA incubation period (24 hrs). VEGF knockdown on mRNA and protein level was highly significant (P < 0.01, t‐test).
© Copyright Policy
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC4940776&req=5

f1: Knockdown efficiency of VEGF on the mRNA and protein level. Left panel: Absolute gene regulation determined by real time quantitative RT‐PCR. Right panel: VEGF protein concentrations determined by ELISA in tissue culture supernatant of cells treated with VEGF siRNA after 24 and 48 hrs after the end of the siRNA incubation period (24 hrs). VEGF knockdown on mRNA and protein level was highly significant (P < 0.01, t‐test).
Mentions: Adherent MSCs in culture flasks were treated for 24 hrs with VEGF siRNA and NeoFX transfection agent in regular growth medium in order to knockdown VEGF expression. Knockdown efficiency was determined at RNA and protein levels 24 hrs and 48 hrs after the end of the transfection period. Efficiency of the knockdown approach was tested before each experiment and adjusted if necessary (combination of siRNAs or different concentrations). Initially, combination of three VEGF siRNA yielded a greater than 80% knockdown of VEGF at the RNA and protein levels. In later experiments, 10 nm of siRNA ID #192613 was used and yielded a greater than 60% knockdown at the protein level. Results are shown in Fig. 1. Knockdown was verified at the mRNA as well as the protein level.

Bottom Line: Knockdown of vascular enthothelial growth factor (VEGF) by siRNA reduced effectiveness of MSCs in the treatment of ischemic AKI in a rat model.Animals treated with MSCs had increased renal microvessel density compared to VEGF knockdown MSC-treated and vehicle-treated animals.These results show that VEGF is an important mediator of the early and late phase of renoprotective action after AKI in the context of stem cell treatment.

View Article: PubMed Central - PubMed

Affiliation: University of Utah, Division of Nephrology, Department of Medicine, Salt Lake City, UT 84148, USA.

ABSTRACT
Adult stem cell treatment of complex disorders is a promising therapeutic approach and multipotent marrow stromal cells (MSCs) have been shown to be effective in various animal models of diseases. Acute kidney injury (AKI) is a common and serious problem in hospitalized patients and bone marrow derived multipotent MSCs have been shown to be effective in different models of AKI. The mechanism of action of MSCs is complex but involves paracrine actions including growth factor secretion. Knockdown of vascular enthothelial growth factor (VEGF) by siRNA reduced effectiveness of MSCs in the treatment of ischemic AKI in a rat model. Animals treated with MSCs had increased renal microvessel density compared to VEGF knockdown MSC-treated and vehicle-treated animals. These results show that VEGF is an important mediator of the early and late phase of renoprotective action after AKI in the context of stem cell treatment.

Show MeSH
Related in: MedlinePlus