脂肪间充质干细胞来源外泌体对缺氧/复氧诱导的心肌细胞凋亡的影响

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摘要目的研究小鼠脂肪间充质干细胞（mASCs）来源外泌体（Exo）对新生小鼠心肌细胞缺氧/复氧（H/R）诱导的心肌细胞凋亡的影响。方法采用超高速离心法分离提取mASCs来源Exo（mASCs-Exo），透射电子显微镜（TEM）、纳米颗粒示踪分析技术（NTA）和蛋白质免疫印迹法（WB）观察并鉴定Exo；差速贴壁法提取C57BL/6乳鼠心肌细胞，免疫荧光检测心肌标志蛋白CTnI和conexin43表达；PKH-26标记Exo示心肌细胞摄取能力；实验分为3组：正常对照组（control组）、缺氧/复氧组（H/R组）和缺氧/复氧+mASCs-Exo组（H/R+mASCs-Exo组）；TUNEL染色检测心肌细胞凋亡；WB检测凋亡相关蛋白表达水平。结果与control组比较，H/R组心肌细胞凋亡率升高（P ＜ 0.01），与H/R组比较，H/R+mASCs-Exo组心肌细胞凋亡率降低（P ＜ 0.05）；与control组比较，H/R组p-PI3K/PI3K、p-Akt/Akt、Bcl-2/Bax比值降低（P ＜ 0.01），而cleaved caspase-9/caspase-9比值升高（P ＜ 0.001）；与H/R组比较，H/R+mASCs-Exo组p-PI3K/PI3K、p-Akt/Akt、Bcl-2/Bax比值升高（P ＜ 0.05或P ＜ 0.01），cleaved caspase-9/caspase-9的比值降低（P ＜ 0.01）。结论 mASCs-Exo可抑制H/R诱导心肌细胞凋亡，这可能通过激活PI3K/Akt信号通路介导的。

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	梁政钟剑锋吴源聪温文陈灿李波

关键词 ：脂肪间充质干细胞, 外泌体, C57BL/6乳鼠心肌细胞, 缺氧/复氧, PI3K/Akt信号通路

Abstract：Objective To investigate the effect of mouse adipose-derived mesenchymal stem cells (mASCs) derived exosomes (Exo) on hypoxia/reoxygenation (H/R) induced apopotosis of cardiomyocytes neonatal mouse cardiomyocytes. Methods The mascs-derived Exo (mascs-exo) was isolated and extracted by ultra-high speed centrifugation. The Exo was observed and identified by transmission electron microscopy (TEM), nanoparticle tracer analysis (NTA) and Western blot (WB). C57BL/6 suckled rat cardiomyocytes were extracted by differential adherent method. Expression of CTnI and conexin43 was detected by immunofluorescence. Pkh-26 was labeled Exo to indicate the uptake capacity of cardiomyocytes. The experiment was divided into three groups: normal control group (control group), hypoxia/reoxygenation group (H/R group) and hypoxia/reoxygenation + mascs-exo group (H/R+ mascs-exo group). Apoptosis of was detected by TUNEL staining. The expression level of apoptosis-related protein was detected by WB. Results compared with the control group, the apoptosis rate of cardiomyocytes in the H/R group was increased (P ＜ 0.01), and the apoptosis rate of cardiomyocytes in the H/R+ mascs-exo group was decreased (P ＜ 0.05). Compared with the control group, p-PI3K /PI3K, p-Akt /Akt, and Bcl-2 /Bax were decreased in the H/R group (P ＜ 0.01), while the cleaved caspase-9/caspase-9 ratio was increased (P ＜ 0.01). Compared with H/R group, the ratio of p-PI3K/PI3K, p-Akt /Akt and Bcl-2 /Bax in H/R+ mascs-exo group was increased (P ＜ 0.05 or P ＜ 0.01), and the ratio of cleaved caspase-9/caspase-9 was decreased (P ＜ 0.01). Conclusion mASCs-Exo can inhibit H/R induction of cardiomyocytes apoptosis, which may be mediated by activation of the PI3K/Akt signaling pathway.

Key words： Adipose-derived mesenchymal stem cells Exosomes Primary cardiomyocytes Hypoxia/reoxygenation PI3K/Akt signaling pathway

基金资助:国家自然科学基金资助项目（81270262）。

通讯作者: 李波（1979.6-），男，博士，副主任医师；研究方向：心肌、缺血再灌注损伤的机制。

作者简介: 梁政（1979.9-），男，硕士，副主任医师；研究方向：心肌缺血再灌注损伤的机制。

引用本文:

梁政钟剑锋吴源聪温文陈灿李波. 脂肪间充质干细胞来源外泌体对缺氧/复氧诱导的心肌细胞凋亡的影响[J]. 中国医药导报, 2020, 17(5): 4-7,22.
LIANG Zheng ZHONG Jianfeng WU Yancong WEN Wen CHEN Can LI Bo. Effect of the exosomes extracted from adipose-derived mesenchymal stem cells on hypoxia/reoxygenation-induced cardiomyocyte apoptosis. 中国医药导报, 2020, 17(5): 4-7,22.

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[1] Benjamin EJ，Blaha MJ，Chiuve SE，et al. Heart Disease and Stroke Statistics-2017 Update：A Report From the American Heart Association [J]. Circulation，2017，135（10）：e146-e603.
[2] Anderson JL，Morrow DA. Acute Myocardial Infarction [J]. N Engl J Med，2017，376（21）：2053-2064.
[3] Patil KD，Halperin HR，Becker LB. Cardiac arrest：resuscitation and reperfusion [J]. Circ Res，2015，116（12）：2041-2049.
[4] Chin KY，Qin C，May L.，et al. New Pharmacological Approaches to the Prevention of Myocardial Ischemia-Reperfusion Injury [J]. Curr Drug Targets，2017，18（15）：1689-1711.
[5] Wernly B，Mirna M，Rezar M，et al. Regenerative Cardiovascular Therapies：Stem Cells and Beyond [J]. Int J Mol Sci，2019，20（6）：1420.
[6] Wei Z，Qiao S，Zhao J，et al. miRNA-181a over-expression in mesenchymal stem cell-derived exosomes influenced inflammatory response after myocardial ischemia-reperfusion injury [J]. Life Sci，2019，232：116632.
[7] Pegtel DM，Gould SJ. Exosomes [J]. Annu Rev Biochem，2019，88：487-514.
[8] Davidson SM，Takov K，Yellon DM. Exosomes and Cardiovascular Protection. Cardiovasc [J]. Drugs Ther，2017， 31：77-86.
[9] Zhang H，Xiang M，Meng D，et al. Inhibition of Myocardial Ischemia/Reperfusion Injury by Exosomes Secreted from Mesenchymal Stem Cells [J]. Stem Cells Int，2016，2016：4328362.
[10] Huang S，Frangogiannis NG. Anti-inflammatory therapies in myocardial infarction：failures，hopes and challenges [J]. Br J Pharmacol，2018，175（9）：1377-1400.
[11] Li Y，Chen B，Yang X，et al. S100a8/a9 Signaling Causes Mitochondrial Dysfunction and Cardiomyocyte Death in Response to Ischemic/Reperfusion Injury [J]. Circulation，2019，140（9）：751-764.
[12] Goradel NH，Hour FG，Negahdari B，et al. Stem Cell Therapy：A New Therapeutic Option for Cardiovascular Diseases [J]. J Cell Biochem，2018，119（1）：95-104.
[13] Golpanian S，Wolf A，Hatzistergos KE，et al. Rebuilding the damaged heart： mesenchymal stem cells，cell-based therapy，and engineered heart tissue [J]. Physiol Rev，2016，96：1127-1168.
[14] Hong HS，Kim S，Lee S，et al. Substance-P Prevents Cardiac Ischemia-Reperfusion Injury by Modulating Stem Cell Mobilization and Causing Early Suppression of Injury-Mediated Inflammation [J]. Cell Physiol Biochem，2019，52（1）：40-56.
[15] Takamura M，Usui S，Inoue O，et al. Adipose-derived regenerative cells exert beneficial effects on systemic responses following myocardial ischemia/reperfusion [J]. Cardiol J，2016，23（6）：685-693.
[16] Vizoso FJ，Eiro N，Cid S，et al. Mesenchymal Stem Cell Secretome：Toward Cell-Free Therapeutic Strategies in Regenerative Medicine [J]. Int J Mol Sci，2017，18（9）Pii：E1852.
[17] Zhao J，Li X，Hu J，et al. Mesenchymal stromal cell-derived exosomes attenuate myocardial ischaemia-reperfusion injurythrough miR-182-regulated macrophage polarization [J]. Cardiovasc Res，2019，115（7）：1205-1216.
[18] Mendt M，Rezvani K，Shpall E. Mesenchymal stem cell-derived exosomes for clinical use [J]. Bone Marrow Transplant，2019，54（Suppl 2）：789-792.
[19] Jovanovi■ A. Cardioprotective signalling： Past，present and future [J]. Eur J Pharmacol，2018，833：314-319.
[20] Shu Z，Yang Y，Yang L，et al. Cardioprotective effects of dihydroquercetin against ischemia reperfusion injury by inhibiting oxidative stress and endoplasmic reticulum stress-induced apoptosis via the PI3K/Akt pathway [J]. Food Funct，2019，10（1）：203-215.