Constitutive Expression of Adiponectin in Endothelial Progenitor Cells Protects a Rat Model of Cerebral Ischemia

Abstract

Endothelial progenitor cells (EPCs), as precursors to endothelial cells, play a significant part in the process of endogenous blood vessel repair and maintenance of endothelial integrity. Adiponectin (APN) is an adipocyte-specific adipocytokine. In this study, we aim to test whether we transplant a combined graft of EPCs transfected with the adiponectin gene into a rat model of cerebral ischemia could improve functional recovery after middle cerebral artery occlusion (MCAO). Sprague-Dawley (SD) rats were randomly divided into a MCAO control group, a MCAO EPC treatment group, and a MCAO LV-APN-EPC treatment group. A focal cerebral ischemia and reperfusion model was induced by the intraluminal suture method. After 2 h of reperfusion, EPCs were transplanted by injection through the tail vein. A rotarod test was conducted to assess behavioral function before MCAO and on days 1, 7, and 14 after MCAO. After 14 d, TTC staining, CD31 immunofluorescence, and TUNEL staining were used to evaluate infarct volume, microvessel density, and cell apoptosis. Results revealed that behavioral function, infarct area percentage, microvessel density, and cell apoptosis rates were more favorable in the LV-APN-EPC treatment group than in the EPC treatment group. These data suggested that gene-modified cell therapy may be a useful approach for the treatment of ischemic stroke.

The Protective Effect of Adiponectin-Transfected Endothelial Progenitor Cells on Cognitive Function in D-Galactose-Induced Aging Rats

“…Changing the media every 72 hours and the growth state of the cells were observed every day. In our previous experiments [27], we identified these cells by the property of uptaking acetylated LDL (ac-LDL) and Ulex europaeus agglutinin-1 (UEA-1) by EPCs, and cells cultured by this method have been identified as EPCs.…”

“…72 h after transfection, the cells were placed under an inverted fluorescence microscope to observe the expression of enhanced green fluorescent protein (EGFP) and evaluate the results of transfection, and the transfection rate was calculated according to the following formula: transfection rate = positive cells/total cells per field × 100%. The expression of APN from APN-EPCs and EPCs were detected by Western blot analysis [27].…”

“…EPCs can produce a variety of cytokines through paracrine to promote the proliferation and differentiation of endothelial cells (ECs) at the site of vascular injury and EPCs in circulation to repair injured vessels under the conditions of chronic inflammation, ischemia, and hypoxia [22][23][24][25][26]. In recent years, EPC transplantation has played an active role in the treatment of ischemic cerebrovascular diseases [25][26][27]. A previous study in our laboratory has also found that the treatment of middle artery occlusion (MCAO) rats with bone marrow-derived EPCs can play a good role in promoting angiogenesis, improving behavioral function, and reducing infarction area and apoptosis rate [27].…”

“…In recent years, EPC transplantation has played an active role in the treatment of ischemic cerebrovascular diseases [25][26][27]. A previous study in our laboratory has also found that the treatment of middle artery occlusion (MCAO) rats with bone marrow-derived EPCs can play a good role in promoting angiogenesis, improving behavioral function, and reducing infarction area and apoptosis rate [27]. Nevertheless, the number of EPCs would decrease with aging [28], and the extraction of EPCs is difficult as well.…”

Adiponectin: a pivotal role in the protection against cerebral ischemic injury

“…Recently, it has been shown that genetically-transplanted endothelial progenitor cells with adiponectin by lentivirus could reduce cerebral infarction volume, improve behavior outcome, increase microvessel density, and reduce cell apoptosis [33] .…”