Apelin combined with bone marrow mesenchymal stem cells transplantation improves cardiac function after myocardial infarction

doi:10.3969/j.issn.2095-4344.0952

Abstract

Abstract:

BACKGROUND: Our previous work demonstrated that apelin could promote bone marrow mesenchymal stem cells (BMSCs) survival and vascularization under hypoxic-ischemic conditions in vitro.
OBJECTIVE: To explore the effect of apelin combined with BMSCs transplantation on cardiac function after myocardial infarction and the relevant mechanisms.
METHODS: Fifty C57BL/6 mice with myocardial infarction were induced by the left anterior descending coronary artery ligation. Animal models were then randomized into five groups: BMSCs, BMSCs+apelin, sh-APJ BMSCs+apelin, apelin and PBS groups. Two weeks after modeling, the mice in the former four groups were subjected to secondary thoracotomy, BMSCs group was given intramyocardial injection of sole BMSCs. BMSCs+apelin and sh-APJ BMSCs+apelin groups were given intramyocardial injection of BMSCs and sh-APJ BMSCs respectively, in combination with intraperitoneal injection of apelin-13 for 2 weeks consecutively. Mice in the PBS group received intramyocardial injection of PBS without BMSCs. Cardiac function of mice was evaluated before and 2 weeks after treatment. MSCs survival and vascularization in the infarct border zone were examined, and the expression of relevant factors was detected thereafter.
RESULTS AND CONCLUSION: (1) At 2 weeks after treatment, the cardiac function of mice was obviously improved in the former four groups in contrast with the baseline, especially in the BMSCs+apelin group; Left ventricular ejection fraction was significantly increased (P < 0.01), and left ventricular end diastolic diameter and left ventricular end systolic diameter were significantly reduced (P < 0.01). (2) Compared with BMSCs and sh-APJ BMSCs+apelin groups, the average optical density of PKH26- and CD31-positive BMSCs in the infarct border zone was significantly enhanced in the BMSCs+apelin group. Moreover, the percentage of CD31 and PKH26 double positive cells to the PKH26-positive BMSCs was considerably increased (P < 0.01). (3) The expression levels of APJ, AKT, pAKT and VEGFA were significantly increased in the BMSCs+apelin group compared with the BMSCs and sh-APJ BMSCs+apelin groups (P < 0.01). However, there was no difference in the above biomarkers between the BMSCs and sh-APJ BMSCs+apelin groups. It was concluded that apelin could combine with APJ to promote BMSCs survival and vascularization in the infarcted myocardial tissues, thereby further improving cardiac function post myocardial infarction. This effect was associated with the up-regulation of VEGFA. Activation of PI3K/AKT signaling pathway was likely to exert some functions in the above procedure.

中国组织工程研究杂志出版内容重点：干细胞；骨髓干细胞；造血干细胞；脂肪干细胞；肿瘤干细胞；胚胎干细胞；脐带脐血干细胞；干细胞诱导；干细胞分化；组织工程

Key words: Bone Marrow, Mesenchymal Stem Cell Transplantation, Myocardial Infarction, Vascular Endothelial Growth Factor A, Tissue Engineering

CLC Number:

R394.2

Hou Jing-ying, Long Hui-bao, Chen Xu-xiang, Wang Lei, Guo Tian-zhu, Zheng Shao-xin, Zhong Ting-ting, Wu Quan-hua, Wu Hao, Wang Tong. Apelin combined with bone marrow mesenchymal stem cells transplantation improves cardiac function after myocardial infarction[J]. Chinese Journal of Tissue Engineering Research, 2018, 22(25): 3937-3943.

Figures/Tables 2

2.1 实验动物数量分析 50只C57BL/6小鼠全部存活，均进入结果分析。 2.2 各组心功能比较小鼠心肌梗死模型建立后进行心脏超声检测，结果显示各组小鼠左室舒张末期内径、左室收缩末期内径、左室舒张末期室间隔厚度、左室收缩末期室间隔厚度、左室射血分数之间差异均无显著性意义(P > 0.05，表1)；在经过相关治疗后2周再次检测心功能，结果显示：与治疗前相比较，MSCs组、MSCs+apelin组、sh-APJ MSCs+apelin组以及apelin组左室舒张末期内径、左室收缩末期内径明显下降，左室射血分数明显增高(P < 0.01，表1和表2)，而PBS组治疗前后相比各指标差异无显著性意义(P > 0.05，表1和表2)。与PBS组相比较，其余各组左室舒张末期内径、左室收缩末期内径明显下降，左室射血分数明显增高(P < 0.01，表2)；与MSCs组、sh-APJ MSCs+apelin组及apelin组相比较，MSCs+apelin组左室舒张末期内径、左室收缩末期内径明显下降，左室射血分数明显增高(P < 0.01，表2)；与MSCs组相比较，apelin组左室舒张末期内径、左室收缩末期内径增加，左室射血分数下降，差异有显著性意义(P < 0.05)。 2.3 MSCs生存和血管再生情况与MSCs组、sh-APJ MSCs+apelin组相比较，MSCs+apelin组梗死边缘区PKH26(+)以及CD31(+)细胞的平均光密度值显著增高(P < 0.01，图1)，PKH26和CD31双阳性细胞占PKH26(+)细胞的比例显著增多(P < 0.01，图1)，差异有显著性意义(P < 0.01，图1)，而sh-APJ MSCs+apelin组与MSCs组相比较，在上述指标方面差异均无显著性意义(P > 0.05)。 2.4 治疗后2周梗死边缘区APJ、AKT、pAKT以及VEGFA表达与PBS组相比较，其他组APJ、AKT、pAKT以及VEGFA表达明显增高(P < 0.01，图2)；与MSCs组和sh-APJ MSCs+apelin组相比，MSCs+apelin组APJ、AKT、pAKT以及VEGFA的蛋白表达水平明显增加(P < 0.01，图2)，而MSCs组、sh-APJ MSCs+apelin组以及apelin组之间上述指标表达差异无显著性意义(P > 0.05，图2)。"

References

[1] Hou J, Zhong T, Guo T, et al. Apelin promotes mesenchymal stem cells survival and vascularization under hypoxic-ischemic condition in vitro involving the upregulation of vascular endothelial growth factor. Exp Mol Pathol. 2017;102(2):203-209.

[2] Narita T, Suzuki K. Bone marrow-derived mesenchymal stem cells for the treatment of heart failure. Heart Fail Rev. 2015;20(1):53-68.

[3] Hou J, Wang L, Hou J, et al. Peroxisome Proliferator-Activated Receptor Gamma Promotes Mesenchymal Stem Cells to Express Connexin43 via the Inhibition of TGF-β1/Smads Signaling in a Rat Model of Myocardial Infarction. Stem Cell Rev. 2015;11(6):885-899.

[4] Karpov AA, Udalova DV, Pliss MG, et al. Can the outcomes of mesenchymal stem cell-based therapy for myocardial infarction be improved? Providing weapons and armour to cells. Cell Prolif. 2017; 50(2). doi: 10.1111/cpr.12316.

[5] Lu L, Wu D, Li L, et al. Apelin/APJ system: A bifunctional target for cardiac hypertrophy. Int J Cardiol. 2017;230:164-170.

[6] Liang D, Han D, Fan W, et al. Therapeutic efficacy of apelin on transplanted mesenchymal stem cells in hindlimb ischemic mice via regulation of autophagy. Sci Rep. 2016;6:21914.

[7] Zeng X, Yu SP, Taylor T, et al. Protective effect of apelin on cultured rat bone marrow mesenchymal stem cells against apoptosis. Stem Cell Res. 2012;8(3):357-367.

[8] Park JS, Yang HN, Yi SW, et al. Neoangiogenesis of human mesenchymal stem cells transfected with peptide-loaded and gene-coated PLGA nanoparticles. Biomaterials. 2016;76:226-237.

[9] 侯婧瑛,汪蕾,钟婷婷,等, apelin干预骨髓间充质干细胞在缺血缺氧条件下的生存和血管再生[J]. 中国组织工程研究,2017,21(1): 6-12.

[10] Lv B, Hua T, Li F,et al. Hypoxia-inducible factor 1 α protects mesenchymal stem cells against oxygen-glucose deprivation-induced injury via autophagy induction and PI3K/AKT/mTOR signaling pathway. Am J Transl Res. 2017;9(5):2492-2499.

[11] Shi XF, Wang H, Xiao FJ, et al. MiRNA-486 regulates angiogenic activity and survival of mesenchymal stem cells under hypoxia through modulating Akt signal. Biochem Biophys Res Commun. 2016;470(3):670-677.

[12] Zou Y, Wang B, Fu W, et al. Apelin-13 Protects PC12 Cells from Corticosterone-Induced Apoptosis Through PI3K and ERKs Activation. Neurochem Res. 2016;41(7):1635-1644.

[13] Zhang J, Liu Q, Hu X, et al. Apelin/APJ signaling promotes hypoxia-induced proliferation of endothelial progenitor cells via phosphoinositide-3 kinase/Akt signaling. Mol Med Rep. 2015;12(3): 3829-3834.

[14] Dicarlo M, Bianchi N, Ferretti C, et al. Evidence Supporting a Paracrine Effect of IGF-1/VEGF on Human Mesenchymal Stromal Cell Commitment. Cells Tissues Organs. 2016;201(5):333-341.

[15] Hou J, Long H, Zhou C, et al. Long noncoding RNA Braveheart promotes cardiogenic differentiation of mesenchymal stem cells in vitro. Stem Cell Res Ther. 2017;8(1):4.

[16] 伍权华,侯婧瑛,郭天柱,等.吡格列酮联合骨髓间充质干细胞移植治疗改善大鼠心肌梗死后的心功能[J].中国组织工程研究, 2015,19(23):3698-3704.

[17] 晏平,侯婧瑛,郑韶欣,等.过氧化物酶体增殖物激活受体γ促进外源性骨髓间充质干细胞表达Cx43的作用及机制干细胞表达Cx43的作用及机制[J].中国组织工程研究, 2016,20(23):3357-3365.

[18] Ji ST, Kim H, Yun J, et al. Promising Therapeutic Strategies for Mesenchymal Stem Cell-Based Cardiovascular Regeneration: From Cell Priming to Tissue Engineering. Stem Cells Int. 2017;2017:3945403.

[19] Li L, Li L, Zhang Z, et al. Hypoxia promotes bone marrow-derived mesenchymal stem cell proliferation through apelin/APJ/autophagy pathway. Acta Biochim Biophys Sin (Shanghai). 2015;47(5):362-367.

[20] Li L, Zeng H, Chen JX. Apelin-13 increases myocardial progenitor cells and improves repair postmyocardial infarction. Am J Physiol Heart Circ Physiol. 2012;303(5):H605-618.

[21] Tempel D, de Boer M, van Deel ED, et al. Apelin enhances cardiac neovascularization after myocardial infarction by recruiting aplnr+ circulating cells. Circ Res. 2012;111(5):585-598.

[22] Kuchroo P, Dave V, Vijayan A, et al. Paracrine factors secreted by umbilical cord-derived mesenchymal stem cells induce angiogenesis in vitro by a VEGF-independent pathway. Stem Cells Dev. 2015;24(4): 437-450.

[23] Rud'ko AS, Efendieva MK, Budzinskaya MV, et al. Influence of vascular endothelial growth factor on angiogenesis and neurogenesis. Vestn Oftalmol. 2017;133(3):75-81.

[24] Crafts TD, Jensen AR, Blocher-Smith EC, et al. Vascular endothelial growth factor: therapeutic possibilities and challenges for the treatment of ischemia. Cytokine. 2015;71(2):385-393.

[25] Pedersen TO, Blois AL, Xue Y, et al. Mesenchymal stem cells induce endothelial cell quiescence and promote capillary formation. Stem Cell Res Ther. 2014;5(1):23.

[26] Mykhaylichenko VY, Kubyshkin AV, Samarin SA, et al. Experimental induction of reparative morphogenesis and adaptive reserves in the ischemic myocardium using multipotent mesenchymal bone marrow-derived stem cells. Pathophysiology. 2016;23(2):95-104.

[27] Tang Y, Gan X, Cheheltani R, et al. Targeted delivery of vascular endothelial growth factor improves stem cell therapy in a rat myocardial infarction model. Nanomedicine. 2014;10(8):1711-1718.

[28] Liu G, Li L, Huo D, et al. A VEGF delivery system targeting MI improves angiogenesis and cardiac function based on the tropism of MSCs and layer-by-layer self-assembly. Biomaterials. 2017;127:117-131.

[29] Ikhapoh IA, Pelham CJ, Agrawal DK. Atherogenic Cytokines Regulate VEGF-A-Induced Differentiation of Bone Marrow-Derived Mesenchymal Stem Cells into Endothelial Cells. Stem Cells Int. 2015;2015:498328.

[30] Xing Y, Hou J, Guo T, et al. microRNA-378 promotes mesenchymal stem cell survival and vascularization under hypoxic-ischemic conditions in vitro. Stem Cell Res Ther. 2014;5(6):130.

[31] Yu JS, Cui W. Proliferation, survival and metabolism: the role of PI3K/AKT/mTOR signalling in pluripotency and cell fate determination. Development. 2016;143(17):3050-3060.

[32] Bader AM, Klose K, Bieback K, et al. Hypoxic Preconditioning Increases Survival and Pro-Angiogenic Capacity of Human Cord Blood Mesenchymal Stromal Cells In Vitro. PLoS One. 2015;10(9):e0138477.

[33] Ma J, Zhao Y, Sun L, et al. Exosomes Derived from Akt-Modified Human Umbilical Cord Mesenchymal Stem Cells Improve Cardiac Regeneration and Promote Angiogenesis via Activating Platelet-Derived Growth Factor D. Stem Cells Transl Med. 2017;6(1):51-59.

[34] Jia X, Pan J, Li X, et al. Bone marrow mesenchymal stromal cells ameliorate angiogenesis and renal damage via promoting PI3k-Akt signaling pathway activation in vivo. Cytotherapy. 2016;18(7):838-845.

[35] Huang C, Dai C, Gong K, et al. Apelin-13 protects neurovascular unit against ischemic injuries through the effects of vascular endothelial growth factor. Neuropeptides. 2016;60:67-74.