Pretreatment of cardiac progenitor cells with bradykinin attenuates H(2)O(2)-induced cell apoptosis and improves cardiac function in rats by regulating autophagy.
Pretreatment of cardiac progenitor cells with bradykinin attenuates H(2)O(2)-induced cell apoptosis and improves cardiac function in rats by regulating autophagy.
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DOI：
10.1186/s13287-021-02503-6
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发表时间：
2021-08-05
影响因子：
7.5
通讯作者：
Li G
中科院分区：
文献类型：
--
作者：
Wu C;Zhou XX;Li JZ;Qiang HF;Wang Y;Li G
既往研究表明，人心脏c-Kit+祖细胞（hCPCs）能有效改善缺血性心脏病。然而，将hCPCs应用于临床治疗的主要挑战是移植hCPCs在宿主心脏中的低存活率，这限制了移植hCPCs的益处。缓激肽（BK）是组织激肽-激肽释放酶系统的主要活性剂。我们前期的研究表明BK通过调节Ca 2+内流介导CPC的生长和迁移。然而，BK对CPCs的保护作用，BK预处理的hCPCs在梗死心脏中的存活率的提高，以及相关的机制仍然是未知的。用H2 O2诱导HCPCs凋亡和自噬，用MTT法、TUNEL法、流式细胞术、Western blotting和mitoSOX法检测不同浓度的BK对H2 O2诱导的HCPCs损伤的修复作用。自噬在BK的抗凋亡作用中的作用被自噬诱导剂雷帕霉素或抑制剂3-甲基腺嘌呤（3-MA）化学激活或抑制。为探讨BK对hCPCs的保护作用，将3-MA或BK预处理的hCPCs移植到心肌梗死大鼠体内。超声心动图用于确定心脏功能，H&E和Masson染色用于评估病理学特征，通过qRT-PCR定量HLA基因表达，并应用免疫染色使用共聚焦显微镜检查新生血管形成。体外实验结果表明，BK通过促进pAkt和Bcl-2的表达，降低caspase 3和Bax的表达，抑制H2 O2诱导的hCPCs凋亡和ROS的产生，并呈浓度依赖性。BK通过降低LC 3 II/I、Beclin 1和ATG 5的表达，增加P62的表达，抑制H2 O2诱导的细胞自噬。在体内实验中，发现与未处理的hCPC或c-kit心肌细胞（CPC细胞）相比，移植的BK或3-MA处理的hCPC通过减少心肌细胞凋亡、炎性浸润和心肌纤维化以及促进梗死心脏中的新血管形成而更有效地改善心脏功能。本研究建立了一种通过BK预处理调控hCPCs细胞凋亡和自噬来挽救梗死区移植hCPCs的新方法，为心力衰竭的治疗提供了一种新的选择。在线版本包含补充材料，可通过10.1186/s13287-021-02503-6获得。
Previous studies have demonstrated that human cardiac c-Kit+ progenitor cells (hCPCs) can effectively improve ischemic heart disease. However, the major challenge in applying hCPCs to clinical therapy is the low survival rate of graft hCPCs in the host heart, which limited the benefit of transplanted hCPCs. Bradykinin (BK) is a principal active agent of the tissue kinin-kallikrein system. Our previous studies have highlighted that BK mediated the growth and migration of CPCs by regulating Ca2+ influx. However, the protective effect of BK on CPCs, improvement in the survival rate of BK-pretreated hCPCs in the infarcted heart, and the related mechanism remain elusive. HCPCs were treated with H2O2 to induce cell apoptosis and autophagy, and different concentration of BK was applied to rescue the H2O2-induced injury detected by MTT assay, TUNEL staining, flow cytometry, western blotting, and mitoSOX assays. The role of autophagy in the anti-apoptotic effect of BK was chemically activated or inhibited using the autophagy inducer, rapamycin, or the inhibitor, 3-methyladenine (3-MA). To explore the protective effect of BK on hCPCs, 3-MA or BK-pretreated hCPCs were transplanted into the myocardial infarcted rats. An echocardiogram was used to determine cardiac function, H&E and Masson staining were employed to assess pathological characteristics, HLA gene expression was quantified by qRT-PCR, and immunostaining was applied to examine neovascularization using confocal microscopy. The in vitro results showed that BK suppressed H2O2-induced hCPCs apoptosis and ROS production in a concentration-dependent manner by promoting pAkt and Bcl-2 expression and reducing cleaved caspase 3 and Bax expression. Moreover, BK restrained the H2O2-induced cell autophagy by decreasing LC3II/I, Beclin1, and ATG5 expression and increasing P62 expression. In the in vivo experiment, the transplanted BK- or 3-MA-treated hCPCs were found to be more effectively improved cardiac function by decreasing cardiomyocyte apoptosis, inflammatory infiltration, and myocardial fibrosis, and promoting neovascularization in the infarcted heart, compared to untreated-hCPCs or c-kit- cardiomyocytes (CPC- cells). Our present study established a new method to rescue transplanted hCPCs in the infarcted cardiac area via regulating cell apoptosis and autophagy of hCPCs by pretreatment with BK, providing a new therapeutic option for heart failure. The online version contains supplementary material available at 10.1186/s13287-021-02503-6.
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影响因子：
5.3
作者：
Che H;Xiao GS;Sun HY;Wang Y;Li GR
通讯作者：
Li GR
影响因子：
6.1
作者：
Liu YY;Sun C;Xue FS;Yang GZ;Li HX;Liu Q;Liao X
通讯作者：
Liao X
DOI：
10.1152/ajpheart.00260.2015
发表时间：
2015-11-15
影响因子：
4.8
作者：
Che, Hui;Li, Gang;Li, Gui-Rong
通讯作者：
Li, Gui-Rong
影响因子：
64.8
作者：
Murry, CE;Soonpaa, MH;Field, LJ
通讯作者：
Field, LJ
DOI：
10.1152/ajpheart.01287.2004
发表时间：
2005-07-01
影响因子：
4.8
作者：
Kintsurashvili, E;Duka, A;Gavras, H
通讯作者：
Gavras, H