Department of Anesthesiology, The Second Affiliated Hospital, Zhejiang University School of Medicine, Hangzhou, 310009, China.
Clinical Skill Training Center, The Second Affiliated Hospital, Zhejiang University School of Medicine, Hangzhou, 310009, China.
Stem Cell Res Ther. 2021 Nov 22;12(1):584. doi: 10.1186/s13287-021-02649-3.
Ischemic heart diseases is one of the leading causes of death worldwide. Although revascularization timely is an effective therapeutic intervention to salvage the ischemic myocardium, reperfusion itself causes additional myocardial injury called ischemia/reperfusion (I/R) injury. Bone marrow-derived mesenchymal stem cells (MSCs) is one of the promising cells to alleviate ischemic myocardial injury. However, this cell therapy is limited by poor MSCs survival after transplantation. Here, we investigated whether sevoflurane preconditioning could promote MSCs to attenuate myocardial I/R injury via transient receptor potential canonical channel 6 (TRPC6)-induced angiogenesis.
The anti-apoptotic effect of sevoflurane preconditioning on MSCs was determined by Annexin V-FITC/propidium iodide staining. TRPC6, hypoxia-inducible factor-1α (HIF-1α), Chemokine receptor 4 (CXCR4) and vascular endothelial growth factor (VEGF) protein expressions and VEGF release from MSCs were determined after hypoxia and reoxygenation (H/R). Small interfering RNA (siRNA) was used to knock down TRPC6 gene expression in MSCs. The angiogenesis of human umbilical vein endothelial cells (HUVECs) co-cultured with MSCs was determined by Matrigel tube formation. Myocardial I/R mouse model was induced by occluding left anterior descending coronary artery for 30 min and then reperfusion. MSCs or sevoflurane preconditioned MSCs were injected around the ligature border zone 5 min before reperfusion. Left ventricle systolic function, infarction size, serum LDH, cTnI and inflammatory cytokines were determined after reperfusion.
Sevoflurane preconditioning up-regulated TRPC6, HIF-1α, CXCR4 and VEGF expressions in MSCs and VEGF release from MSCs under H/R, which were reversed by knockdown of TRPC6 gene using siRNA in MSCs. Furthermore, sevoflurane preconditioning promoted the angiogenic and anti-inflammatory effect of HUVECs co-cultured with MSCs. Sevoflurane preconditioned MSCs improved left ventricle systolic function and alleviated myocardial infarction and inflammation in mice subjected to I/R insult.
The current findings reveal that sevoflurane preconditioned MSCs boost angiogenesis in HUVECs subjected to H/R insult and attenuate myocardial I/R injury, which may be mediated by TRPC6 up-regulated HIF-1α, CXCR4 and VEGF.
缺血性心脏病是全球范围内主要的死亡原因之一。尽管及时进行血运重建是挽救缺血心肌的有效治疗干预措施，但再灌注本身会导致缺血/再灌注（I/R）损伤，即额外的心肌损伤。骨髓间充质干细胞（MSCs）是一种有前途的细胞，可以减轻缺血性心肌损伤。然而，这种细胞疗法受到移植后 MSCs 存活率低的限制。在这里，我们研究了七氟醚预处理是否可以通过瞬时受体电位经典通道 6（TRPC6）诱导的血管生成来促进 MSCs 减轻心肌 I/R 损伤。
通过 Annexin V-FITC/碘化丙啶染色确定七氟醚预处理对 MSCs 的抗凋亡作用。在缺氧和再氧合（H/R）后，确定 MSCs 中的 TRPC6、缺氧诱导因子-1α（HIF-1α）、趋化因子受体 4（CXCR4）和血管内皮生长因子（VEGF）蛋白表达和 VEGF 释放。用小干扰 RNA（siRNA）敲低 MSCs 中的 TRPC6 基因表达。用人脐静脉内皮细胞（HUVEC）与 MSCs 共培养来确定血管生成。通过结扎左前降支 30 分钟然后再灌注来诱导心肌 I/R 小鼠模型。在再灌注前 5 分钟将 MSCs 或七氟醚预处理的 MSCs 注射到结扎边界区周围。再灌注后测定左心室收缩功能、梗死面积、血清 LDH、cTnI 和炎症细胞因子。
七氟醚预处理上调了 H/R 条件下 MSCs 中的 TRPC6、HIF-1α、CXCR4 和 VEGF 表达以及 MSCs 中 VEGF 的释放，而用 MSCs 中的 siRNA 敲低 TRPC6 基因则逆转了这种作用。此外，七氟醚预处理促进了与 MSCs 共培养的 HUVEC 的血管生成和抗炎作用。七氟醚预处理的 MSCs 改善了 I/R 损伤小鼠的左心室收缩功能并减轻了心肌梗死和炎症。
目前的研究结果表明，七氟醚预处理的 MSCs 增强了 H/R 应激下 HUVEC 的血管生成，并减轻了心肌 I/R 损伤，这可能是通过上调的 TRPC6 介导的 HIF-1α、CXCR4 和 VEGF 实现的。
