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Up‐regulation of Clusterin is Involved in MSC Overexpressing GATA‐4 Mediated Cell Protection
Author(s) -
Yang Yueting,
Yu Bin,
Liu Huan,
Wang YiGang,
Ashraf Muhammad,
Xu Meifeng
Publication year - 2015
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.29.1_supplement.978.3
Subject(s) - clusterin , cytoprotection , mesenchymal stem cell , downregulation and upregulation , transfection , microbiology and biotechnology , apoptosis , blot , cell , ly294002 , biology , cancer research , chemistry , protein kinase b , andrology , medicine , cell culture , signal transduction , gene , biochemistry , genetics
We have previously reported that GATA‐4 gene transfection in mesenchymal stem cells (MSC) prolongs MSC survival in the ischemic heart and promotes heart repair. Our preliminary studies show that clusterin was prominently upregulated in MSCs overexpressing GATA‐4 (MSC GATA‐4 ). In this study, we hypothesized that upregulation of clusterin was responsible for GATA‐4 mediated cell protection. Methods and Results MSCs were harvested from rat bone marrow and transduced with GATA‐4 by using the murine stem cell virus retroviral expression system. The expression of clusterin in MSCs was documented by real‐time PCR, immunofluorescence, and western blotting. The released clusterin from MSCs was estimated using ELISA. Cardiomyocytes (CM) from neonatal rat ventricles were cultured and exposed to hypoxia and anoxia to mimic ischemic injury. The cytoprotection was determined by measuring lactate dehydrogenase release, MTT intake, Annexin‐V staining, and mitochondrial membrane potential (ΔΨm). Compared to vector‐empty transduced control MSCs, the expression of clusterin was significantly up‐regulated in MSC GATA‐4 accompanied by its significant release from MSC GATA‐4 . Direct administration of clusterin protected MSCs and CM against ischemic injury by increased cell survival, reduced apoptosis, and maintenance of ΔΨm. The cytoprotective effect of clusterin was partially abrogated by Akt inhibitor LY294002. Clusterin also significantly augmented expression of p‐Akt in MSCs and CM. Conclusions It is concluded that clusterin played a prominent role in MSC GATA‐4 mediated cytoprotection via Akt signaling.