Mesenchymal stem cell-conditioned media ameliorate diabetic endothelial dysfunction by improving mitochondrial bioenergetics via the Sirt1/AMPK/PGC-1α pathway.
Vasculopathy is a major complication of diabetes. Impaired mitochondrial bioenergetics and biogenesis due to oxidative stress are a critical causal factor for diabetic endothelial dysfunction. Sirt1, an NAD+-dependent enzyme, is known to play an important protective role through deacetylation of many substrates involved in oxidative phosphorylation and reactive oxygen species generation. Mesenchymal stem cell-conditioned medium (MSC-CM) has emerged as a promising cell-free therapy due to the trophic actions of mesenchymal stem cell (MSC)-secreted molecules. In the present study, we investigated the therapeutic potential of MSC-CMs in diabetic endothelial dysfunction, focusing on the Sirt1 signalling pathway and the relevance to mitochondrial function. We found that high glucose-stimulated MSC-CM attenuated several glucotoxicity-induced processes, oxidative stress and apoptosis of endothelial cells of the human umbilical vein. MSC-CM perfusion in diabetic rats ameliorated compromised aortic vasodilatation and alleviated oxidative stress in aortas. We further demonstrated that these effects were dependent on improved mitochondrial function and up-regulation of Sirt1 expression. MSC-CMs activated the phosphorylation of phosphoinositide 3-kinase (PI3K) and protein kinase B (Akt), leading to direct interaction between Akt and Sirt1, and subsequently enhanced Sirt1 expression. In addition, both MSC-CM and Sirt1 activation could increase the expression of peroxisome proliferator-activated receptor γ co-activator-1α (PGC-1α), as well as increase the mRNA expression of its downstream, mitochondrial, biogenesis-related genes. This indirect regulation was mediated by activation of AMP-activated protein kinase (AMPK). Overall our findings indicated that MSC-CM had protective effects on endothelial cells, with respect to glucotoxicity, by ameliorating mitochondrial dysfunction via the PI3K/Akt/Sirt1 pathway, and Sirt1 potentiated mitochondrial biogenesis, through the Sirt1/AMPK/PGC-1α pathway.
Key Laboratory of Transplant Engineering and Immunology, NHFPC; Regenerative Medicine Research Centre, West China Hospital, Sichuan University, Chengdu, People's Republic of China.
Yujia
Y
Yuan
Key Laboratory of Transplant Engineering and Immunology, NHFPC; Regenerative Medicine Research Centre, West China Hospital, Sichuan University, Chengdu, People's Republic of China.
Meimei
M
Shi
Key Laboratory of Transplant Engineering and Immunology, NHFPC; Regenerative Medicine Research Centre, West China Hospital, Sichuan University, Chengdu, People's Republic of China.
Lan
L
Li
Key Laboratory of Transplant Engineering and Immunology, NHFPC; Regenerative Medicine Research Centre, West China Hospital, Sichuan University, Chengdu, People's Republic of China.
Jingping
J
Liu
Key Laboratory of Transplant Engineering and Immunology, NHFPC; Regenerative Medicine Research Centre, West China Hospital, Sichuan University, Chengdu, People's Republic of China.
Bo
B
Chen
Key Laboratory of Transplant Engineering and Immunology, NHFPC; Regenerative Medicine Research Centre, West China Hospital, Sichuan University, Chengdu, People's Republic of China.|School of Biomedical Sciences, CHIRI Biosciences, Curtin University, GPO Box U1987, Perth, Western Australia, Australia.
Younan
Y
Chen
Key Laboratory of Transplant Engineering and Immunology, NHFPC; Regenerative Medicine Research Centre, West China Hospital, Sichuan University, Chengdu, People's Republic of China.
Xingxing
X
An
Key Laboratory of Transplant Engineering and Immunology, NHFPC; Regenerative Medicine Research Centre, West China Hospital, Sichuan University, Chengdu, People's Republic of China.
Shuyun
S
Liu
Key Laboratory of Transplant Engineering and Immunology, NHFPC; Regenerative Medicine Research Centre, West China Hospital, Sichuan University, Chengdu, People's Republic of China.
Ruixi
R
Luo
Key Laboratory of Transplant Engineering and Immunology, NHFPC; Regenerative Medicine Research Centre, West China Hospital, Sichuan University, Chengdu, People's Republic of China.
Dan
D
Long
Key Laboratory of Transplant Engineering and Immunology, NHFPC; Regenerative Medicine Research Centre, West China Hospital, Sichuan University, Chengdu, People's Republic of China.
Wengeng
W
Zhang
School of Biomedical Sciences, CHIRI Biosciences, Curtin University, GPO Box U1987, Perth, Western Australia, Australia.
Philip
P
Newsholme
Key Laboratory of Transplant Engineering and Immunology, NHFPC; Regenerative Medicine Research Centre, West China Hospital, Sichuan University, Chengdu, People's Republic of China.
Jingqiu
J
Cheng
Key Laboratory of Transplant Engineering and Immunology, NHFPC; Regenerative Medicine Research Centre, West China Hospital, Sichuan University, Chengdu, People's Republic of China luyanrong@scu.edu.cn.
Yanrong
Y
Lu
D055372:AMP-Activated Protein Kinases / Q000235:genetics / Q000378:metabolism*; D000818:Animals; D017209:Apoptosis; D017077:Culture Media, Conditioned / Q000378:metabolism / Q000494:pharmacology*; D003921:Diabetes Mellitus, Experimental / Q000235:genetics / Q000378:metabolism / Q000503:physiopathology / Q000628:therapy*; D005947:Glucose / Q000378:metabolism; D061307:Human Umbilical Vein Endothelial Cells / Q000166:cytology / Q000378:metabolism; D006801:Humans; D008297:Male; D059630:Mesenchymal Stem Cells / Q000166:cytology / Q000378:metabolism*; D008928:Mitochondria / Q000235:genetics / Q000378:metabolism*; D018384:Oxidative Stress; D000071248:Peroxisome Proliferator-Activated Receptor Gamma Coactivator 1-alpha / Q000235:genetics / Q000378:metabolism*; D051381:Rats; D017207:Rats, Sprague-Dawley; D015398:Signal Transduction; D056564:Sirtuin 1 / Q000235:genetics / Q000378:metabolism*
D017077:Culture Media, Conditioned; C486497:PPARGC1A protein, human; D000071248:Peroxisome Proliferator-Activated Receptor Gamma Coactivator 1-alpha; D055372:AMP-Activated Protein Kinases; D056564:Sirtuin 1; D005947:Glucose
endothelial dysfunction; mesenchymal stem cell-conditioned media; mitochondrial biogenesis; sirtuins