Department of Anesthesiology, Beijing Anzhen Hospital, Capital Medical University, Beijing 100029, China.
Department of Anesthesiology, Affiliated Hospital of Chengde Medical College, Chengde 067000, China.
Comb Chem High Throughput Screen. 2023;26(6):1204-1213. doi: 10.2174/1386207325666220715090505.
The lack of effective treatments for myocardial ischemiareperfusion (MI-R) injury severely restricts the effectiveness of the treatment of ischemic heart disease. In the present research, we aimed to investigate the protective effect and molecular mechanism of penehyclidine hydrochloride (PHC) on MI-R cells.
Cell viability was quantified using CCK8. Cell apoptosis was analyzed using flow cytometry. Western blot and Elisa assays were used for the detection of target proteins.
PHC pretreatment attenuated the inhibition of cell viability and decreased the percentage of apoptosis induced by simulated ischemia reperfusion (SIR). Platelet-derived growth factor B (PDGF-B) and its downstream AKT pathway were activated in PHC pretreated cells. After siRNAPDGF- B transfection, cell viability was inhibited and apoptosis was activated in PHC pretreated SIR cells, suggesting that PHC protected cells from SIR. PDGF-B knockdown also increased the levels of CK, LDH, IL-6 and TNF-α in PHC pretreated SIR cells. The effect of AKT inhibitor on H9C2 cells was consistent with that of PDGF-B knockdown.
PHC pretreatment can protect cardiomyocytes from the decrease of cell activity and the increase of apoptosis caused by reperfusion through up-regulating PDGF-B to activate PI3K pathway. Our study indicates that PHC is a potential drug to protect cells from reperfusion injury and PDGF-B is a potential target for preventing MI-R injury.
心肌缺血再灌注（MI-R）损伤缺乏有效治疗方法，严重限制了缺血性心脏病治疗的效果。在本研究中，我们旨在研究盐酸戊乙奎醚（PHC）对 MI-R 细胞的保护作用及其分子机制。
用 CCK8 定量检测细胞活力。用流式细胞术分析细胞凋亡。用 Western blot 和 Elisa 检测法检测靶蛋白。
PHC 预处理可减轻模拟缺血再灌注（SIR）引起的细胞活力抑制和凋亡百分比增加。血小板衍生生长因子 B（PDGF-B）及其下游 AKT 通路在 PHC 预处理细胞中被激活。在 siRNAPDGF-B 转染后，PHC 预处理 SIR 细胞中的细胞活力受到抑制，细胞凋亡被激活，表明 PHC 可保护细胞免受 SIR 损伤。PDGF-B 敲低也增加了 PHC 预处理 SIR 细胞中 CK、LDH、IL-6 和 TNF-α的水平。AKT 抑制剂对 H9C2 细胞的作用与 PDGF-B 敲低的作用一致。
PHC 预处理可通过上调 PDGF-B 激活 PI3K 通路，保护心肌细胞免受再灌注引起的细胞活性下降和凋亡增加。我们的研究表明，PHC 是一种保护细胞免受再灌注损伤的潜在药物，PDGF-B 是预防 MI-R 损伤的潜在靶点。
