Extracorporeal Cardiac Shock Wave-Induced Exosome Derived From Endothelial Colony-Forming Cells Carrying miR-140-3p Alleviate Cardiomyocyte Hypoxia/Reoxygenation Injury via the PTEN/PI3K/AKT Pathway.
Extracorporeal Cardiac Shock Wave-Induced Exosome Derived From Endothelial Colony-Forming Cells Carrying miR-140-3p Alleviate Cardiomyocyte Hypoxia/Reoxygenation Injury via the PTEN/PI3K/AKT Pathway.
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体外心脏冲击波诱导的源自携带 miR-140-3p 的内皮集落形成细胞的外泌体通过 PTEN/PI3K/AKT 途径减轻心肌细胞缺氧/复氧损伤
DOI：
10.3389/fcell.2021.779936
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发表时间：
2021
影响因子：
5.5
通讯作者：
Cai H
中科院分区：
文献类型：
--
作者：
Yang D;Wang M;Hu Z;Ma Y;Shi Y;Cao X;Guo T;Cai H;Cai H
背景：干细胞来源的外泌体在心肌缺血再灌注损伤（IRI）的治疗中具有巨大的潜力。体外心脏冲击波（ECSW）作为一种有效的治疗方法，可以部分激活exosomes的功能。本研究旨在探讨ECSW诱导的内皮集落形成细胞来源的exosome对心肌细胞缺氧/复氧（H/R）损伤的影响及其机制。 方法：从内皮细胞集落形成细胞（ECFCs-exo）培养上清中提取并纯化exosomes。采用高通量测序方法对经冲击波（SW-exo）或未经冲击波（CON-exo）处理的ECFCs-exo进行miRNA测序。H9 c2细胞在H/R损伤后与SW-exo或CON-exo孵育。检测细胞活力、细胞凋亡、氧化应激水平和炎症因子。采用qRT-PCR检测细胞和exosomes中miRNA和mRNA的表达水平。Western blotting检测PTEN/PI 3 K/AKT通路相关蛋白表达。 结果：ECFC分泌的Exosomes可被H9 c2细胞摄取。SW-exo可显著提高H/R损伤后H9 c2细胞的存活率，抑制细胞凋亡，下调细胞氧化应激水平（p < 0.01），增加Bcl-2蛋白表达，降低Bax、caspase-3和NF-κB蛋白表达（p < 0.05）。值得注意的是，发现miR-140- 3 p在用ECSW处理的ECFC和ECFC-exo中高度富集（p < 0.05），并充当关键介体。SW-exo增加了H9 c2细胞中miR-140- 3 p的表达，但降低了PTEN的表达，同时增强了PI 3 K/AKT信号通路的磷酸化。SW-exo对H/R损伤的这些心脏保护作用被miR-140- 3 p抑制剂减弱。双荧光素酶实验证实miR-140- 3 p可直接靶向PTEN mRNA的3′UTR，发挥负调控作用。 结论：本研究表明，ECSW作为一种有效的刺激ECFC来源的exosomes在体外的潜力。SW-exo对H9 c2细胞H/R损伤有较强的治疗作用，其机制可能与转染exosomal miR-140- 3 p有关。
Background: Stem cell-derived exosomes have great potential in the treatment of myocardial ischemia–reperfusion injury (IRI). Extracorporeal cardiac shock waves (ECSW) as effective therapy, in part, could activate the function of exosomes. In this study, we explored the effect of ECSW-induced exosome derived from endothelial colony-forming cells on cardiomyocyte hypoxia/reoxygenation (H/R) injury and its underlying mechanisms. Methods: The exosomes were extracted and purified from the supernatant of endothelial colony-forming cells (ECFCs-exo). ECFCs-exo treated with shock wave (SW-exo) or without shock wave (CON-exo) were performed with high-throughput sequencing of the miRNA. H9c2 cells were incubated with SW-exo or CON-exo after H/R injury. The cell viability, cell apoptosis, oxidative stress level, and inflammatory factor were assessed. qRT-PCR was used to detect the expression levels of miRNA and mRNA in cells and exosomes. The PTEN/PI3K/AKT pathway-related proteins were detected by Western blotting, respectively. Results: Exosomes secreted by ECFCs could be taken up by H9c2 cells. Administration of SW-exo to H9c2 cells after H/R injury could significantly improve cell viability, inhibit cell apoptosis, and downregulate oxidative stress level (p < 0.01), with an increase in Bcl-2 protein and a decrease in Bax, cleaved caspase-3, and NF-κB protein (p < 0.05). Notably, miR-140-3p was found to be highly enriched both in ECFCs and ECFCs-exo treated with ECSW (p < 0.05) and served as a critical mediator. SW-exo increased miR-140-3p expression but decreased PTEN expression in H9c2 cells with enhanced phosphorylation of the PI3K/AKT signaling pathway. These cardioprotective effects of SW-exo on H/R injury were blunted by the miR-140-3p inhibitor. Dual-luciferase assay verified that miR-140-3p could directly target the 3′UTR of PTEN mRNA and exert a negative regulatory effect. Conclusion: This study has shown the potential of ECSW as an effective stimulation for the exosomes derived from ECFCs in vitro. SW-exo exerted a stronger therapeutic effect on H/R injury in H9c2 cells possibly via delivering exosomal miR-140-3p, which might be a novel promising strategy for the myocardial IRI.
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影响因子：
3.7
作者：
Feng Y;Huang W;Wani M;Yu X;Ashraf M
通讯作者：
Ashraf M
DOI：
10.1136/heartjnl-2015-307855
发表时间：
2016-03
期刊：
Heart (British Cardiac Society)
影响因子：
--
作者：
Bulluck H;Yellon DM;Hausenloy DJ
通讯作者：
Hausenloy DJ
PTEN 通过 PI3K/AKT 信号通路介导 H9c2 细胞中血清剥夺诱导的细胞毒性
影响因子：
3.2
作者：
Gong, Mengmeng;Li, Zhiqiang;Wei, Yidong
通讯作者：
Wei, Yidong
PTEN抑制剂通过PI3k/Akt/VEGF信号通路改善心肌梗死后血管重塑和心功能
DOI：
10.1186/s10020-020-00241-8
发表时间：
2020-11-19
期刊：
Molecular medicine (Cambridge, Mass.)
影响因子：
--
作者：
Feng Q;Li X;Qin X;Yu C;Jin Y;Qian X
通讯作者：
Qian X
影响因子：
9
作者：
Ghosh A;Dasgupta D;Ghosh A;Roychoudhury S;Kumar D;Gorain M;Butti R;Datta S;Agarwal S;Gupta S;Krishna Dhali G;Chowdhury A;Schmittgen TD;Kundu GC;Banerjee S
通讯作者：
Banerjee S