Apelin干预骨髓间充质干细胞移植治疗改善心肌梗死后心功能
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摘要
背景:课题组前期研究发现血管紧张素受体AT1相关的受体蛋白内源性配体(apelin)能够促进骨髓间充质干细胞(mesenchymal stem cells,MSCs)在体外缺血缺氧条件下的生存和血管再生。目的:采用体内实验探讨apelin联合MSCs移植治疗对心肌梗死后心功能的影响及相关机制。方法:开胸结扎50只C57BL/6小鼠左前降支冠状动脉并随机分为5组:MSCs组、MSCs+apelin组、sh-APJ MSCs+apelin组、apelin组和PBS组。模型建立2周后,对动物进行第2次开胸手术,MSCs组于局部梗死心肌内注射单纯的MSCs,MSCs+apelin组和sh-APJ MSCs+apelin组分别注射MSCs以及sh-APJ MSCs并且均连续腹腔内注射2周apelin-13,PBS组心肌内注射不含MSCs的PBS。各组治疗前及治疗2周后检测心功能。实验结束后,摘取心脏检测左心室心肌组织梗死边缘区的MSCs生存和血管再生情况以及相关因子的表达。结果与结论:(1)与治疗前比较,治疗2周后MSCs组、MSCs+apelin组、sh-APJ MSCs+apelin组、apelin组左室射血分数明显增加(P<0.01),左室舒张末期内径、左室收缩末期内径显著减小(P<0.01);其中MSCs+apelin组以上指标改善最显著(P<0.01);(2)与MSCs组和sh-APJ MSCs+apelin组相比,MSCs+apelin组梗死边缘区PKH26阳性细胞和CD31阳性细胞的平均光密度值显著增高,且PKH26和CD31双阳性细胞占PKH26阳性细胞的比例显著增加(P<0.01);(3)与MSCs组和sh-APJ MSCs+apelin组相比较,MSCs+apelin组APJ、AKT、p AKT以及VEGFA表达显著增加,而MSCs组和sh-APJ MSCs+apelin组在上述各项指标表达方面差异均无显著性意义(P>0.05);(4)以上提示apelin能够通过与其受体APJ结合改善MSCs在心肌梗死局部组织中的生存和血管再生,从而进一步提高心肌梗死后心功能,此效应与VEGFA表达上调相关,PI3K/AKT的激活可能在其中起到一定的调控作用。
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 C57 BL/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, p AKT 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 PI3 K/AKT signaling pathway was likely to exert some functions in the above procedure.
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 C57 BL/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, p AKT 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 PI3 K/AKT signaling pathway was likely to exert some functions in the above procedure.
引文
[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 1αprotects mesenchymal stem cells against oxygen-glucose deprivation-induced injury via autophagy induction and PI3K/AKT/m TOR signaling pathway.Am J Transl Res.2017;9(5):2492-2499.
[11]Shi XF,Wang H,Xiao FJ,et al.Mi RNA-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.micro RNA-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/m TOR 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.PLo S 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.
[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 1αprotects mesenchymal stem cells against oxygen-glucose deprivation-induced injury via autophagy induction and PI3K/AKT/m TOR signaling pathway.Am J Transl Res.2017;9(5):2492-2499.
[11]Shi XF,Wang H,Xiao FJ,et al.Mi RNA-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.micro RNA-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/m TOR 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.PLo S 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.