Allicin protects against myocardial I/R by accelerating angiogenesis via the miR-19a-3p/PI3K/AKT axis.
Allicin protects against myocardial I/R by accelerating angiogenesis via the miR-19a-3p/PI3K/AKT axis.
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大蒜素通过 miR-19a-3p/PI3K/AKT 轴加速血管生成，预防心肌 I/R
DOI：
10.18632/aging.203578
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发表时间：
2021-10-04
期刊：
影响因子：
--
通讯作者：
Li X
中科院分区：
文献类型：
--
作者：
Liu M;Yang P;Fu D;Gao T;Deng X;Shao M;Liao J;Jiang H;Li X
目的：大蒜素是一种烯丙基 2-丙烯硫代亚磺酸盐或二烯丙基硫代亚磺酸盐，对心肌缺血/再灌注 (MI/R) 损伤具有心脏保护作用。本研究旨在探讨大蒜素预防 MI/R 的潜在机制。方法：对C57BL6小鼠进行假手术或MI/R手术，大蒜素组小鼠在诱导缺血前注射大蒜素（5 mg/ml）。通过超声定量和Masson染色评价实验小鼠的心功能和组织病理学。接下来，我们通过马森染色和免疫组织化学染色测量了梗死周围区域的毛细血管生成。采用miRNA芯片检测MI/R组织和相应正常组织中表达的miRNA。进行实时定量聚合酶链反应（q-PCR）以验证所选的 miRNA-19α-3p 基因表达。此外，我们通过免疫荧光染色评估了心肌乳酸脱氢酶和COX-2。 Western blot分析评估大蒜素组和模型组p-AKT、p-PI3K、p-mTOR、COX-2和VEGF蛋白水平。在体外研究中，LPS 刺激表达 Tie2 的巨噬细胞在缺血缓冲液中培养。我们通过 fura-2/AM 荧光评估了 VEGF 的积累。此外，进行Western blotting检测p-PI3K、p-AKT、p-mTOR、VEGF、COX2和MMP2的蛋白水平。应用 PI3K 抑制剂来研究大蒜素诱导的心肌缺血再灌注损伤保护是否是通过 PI3K/AKT 通路介导的。并转染miR-19α-3p模拟物/抑制剂来促进/抑制miR-19a-3p的表达，以验证miR-19a-3p对PI3K通路的调节作用。结果：大蒜素预处理显着改善缺血再灌注引起的心功能损伤。此外，大蒜素可以抑制心脏纤维化，心脏纤维化面积的减少就证明了这一点。大蒜素对 MI/R 的影响与毛细血管生成的增加有关。微阵列分析表明，miR-19a-3p 通过直接靶向 PIK3CA 基因下调 PIK3CA (PI3K) 表达。血管生成途径和基因 miRNA-19a-3p 的调节可能会影响大蒜素诱导的 MI/R 保护。免疫荧光染色显示，大蒜素处理后COX-2和心肌乳酸脱氢酶显着升高。此外，蛋白质印迹分析表明，大蒜素组中 p-AKT、p-PI3K、p-mTOR、COX-2 和 VEGF 蛋白水平也有所增加。体外研究发现，经大蒜素处理的表达 Tie2 的巨噬细胞中 p-PI3K、p-AKT、p-mTOR、VEGF、COX2 和 MMP2 的蛋白水平显着增加。 PI3K 抑制剂（Wortmannin）治疗可部分逆转这些影响。 MiR-19α-3p在心肌I/R损伤中发挥重要作用。它可以调节 PI3K-AKT 通路的活性。抑制miR-19a-3p可通过调节PI3K/AKT通路促进血管生成。结论：大蒜素预处理可预防心肌缺血再灌注并激活 miR-19a-3p/PI3K/AKT 通路。
Objectives: Allicin is an allyl 2-propenethiosulfinate or diallyl thiosulfinate acid with cardioprotective effects in myocardial ischemia/reperfusion (MI/R) injury. This study aims to examine the underlying mechanism by which Allicin protects against MI/R. Methods: C57BL6 mice were subjected to either sham or MI/R surgery, and mice in the Allicin group were injected with Allicin (5 mg/ml) before the induction of ischemia. The cardiac function and histopathology of experimental mice were evaluated by ultrasound quantification and Masson staining. We next measured the capillary angiogenesis of the peri-infarct area by Masson staining and immunohistochemical staining. The miRNA microarray was carried out to examine the expressed miRNAs in MI/R tissues and corresponding normal tissues. Real-time quantitative polymerase chain reaction (q-PCR) was performed to validate the selected miRNA-19α-3p gene expression. Besides, we evaluated the myocardial lactate dehydrogenase and COX-2 by immunofluorescence staining. The western blot analysis was used to evaluate the protein levels of p-AKT, p-PI3K, p-mTOR, COX-2, and VEGF protein in the Allicin and Model group. In vitro study, LPS stimulated Tie2 expressing macrophages were cultured in an ischemic buffer. We evaluated the accumulation of VEGF by fura-2/AM fluorescence. Besides, Western blotting was performed to examine the protein levels of p-PI3K, p-AKT, p-mTOR, VEGF, COX2, and MMP2. The PI3K inhibitor was applied to investigate whether Allicin-induced myocardial ischemia-reperfusion injury protection is mediated via the PI3K/AKT pathway. And the miR-19α-3p mimic/inhibitor were transfected to promote/inhibit the expression of miR-19a-3p for verifying the regulation of miR-19a-3p on PI3K pathway. Results: Allicin pretreatment significantly improved I/R-induced cardiac function damage. Furthermore, Allicin could repress cardiac fibrosis, as evidenced by reduced areas of cardiac fibrosis. Allicin’s effect on the MI/R was associated with increased capillary angiogenesis. Microarray analysis exposed that miR-19a-3p down-regulated PIK3CA (PI3K) expression by directly targeting the PIK3CA gene. The regulation of the angiogenesis pathway and gene miRNA-19a-3p might affect the Allicin-induced MI/R protection. Immunofluorescence staining revealed that COX-2 and myocardial lactate dehydrogenase were significantly increased after Allicin treatment. Furthermore, western blot analysis demonstrated that p-AKT, p-PI3K, p-mTOR, COX-2, and VEGF protein levels were also increased in the Allicin group. In vitro study, the protein levels of p-PI3K, p-AKT, p-mTOR, VEGF, COX2, and MMP2 were significantly increased in the Allicin-treated Tie2 expressing macrophages. These effects were partially reversed by PI3K inhibitor (Wortmannin) treatment. MiR-19α-3p plays an important role in myocardial I/R injury. It could regulate the activity of the PI3K-AKT pathway. And inhibition of miR-19a-3p promoted angiogenesis by regulating PI3K/AKT pathway. Conclusions: Allicin pretreatment protects against myocardial I/R and activating the miR-19a-3p/PI3K/AKT pathway.
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DOI：
10.1155/2020/8539148
发表时间：
2020
期刊：
Evidence-based complementary and alternative medicine : eCAM
影响因子：
--
作者：
Liu M;Xue Y;Liang Y;Xue Y;Han X;Li Z;Chu L
通讯作者：
Chu L
抑制动力相关蛋白 1 可预防糖尿病小鼠心肌缺血再灌注损伤
影响因子：
9.3
作者：
Ding M;Dong Q;Liu Z;Liu Z;Qu Y;Li X;Huo C;Jia X;Fu F;Wang X
通讯作者：
Wang X
DOI：
10.7534/j.issn.1009-2137.2015.04.052
发表时间：
2015-08-01
期刊：
Zhongguo shi yan xue ye xue za zhi
影响因子：
--
作者：
Wang, Xiao-Xue;Wang, Yue;Li, Yan
通讯作者：
Li, Yan
影响因子：
4.5
作者：
Manku, Gurpreet;Papadopoulos, Philippos;Culty, Martine
通讯作者：
Culty, Martine
影响因子：
3.6
作者：
Charles CJ;Li RR;Yeung T;Mazlan SMI;Lai RC;de Kleijn DPV;Lim SK;Richards AM
通讯作者：
Richards AM