Department of Anesthesiology, The First Affiliated Hospital of China Medical University, Shenyang, Liaoning 110016, P.R. China.
Department of Clinical Laboratory, General Hospital of Shenyang Military Area Command, Shenyang, Liaoning 110016, P.R. China.
Mol Med Rep. 2018 Aug;18(2):1925-1938. doi: 10.3892/mmr.2018.9198. Epub 2018 Jun 20.
Myocardial ischemia, hypoxia and reperfusion injury are induced by aortic occlusion, cardiac arrest and resuscitation during cardiopulmonary bypass (CPB), which can severely affect cardiac function. The aim of the present study was to investigate the effects of hydrogen‑rich solution (HRS) and aquaporin (AQP) on cardiopulmonary bypass (CPB)‑induced myocardial injury, and determine the mechanism of the phosphatidylinositol 3‑kinase (PI3K)/protein kinase B (Akt) signaling pathway. Sprague Dawley rats were divided into a sham operation group, a CPB surgery group and a HRS group. A CPB model was established, and the hemodynamic parameters were determined at the termination of CPB. The myocardial tissues were observed by hematoxylin and eosin, and Masson staining. The levels of myocardial injury markers [adult cardiac troponin I (cTnI), lactate dehydrogenase (LDH), creatine kinase MB (CK‑MB) and brain natriuretic peptide (BNP)], inflammatory factors [interleukin (IL)‑1β, IL‑6 and tumor necrosis factor‑α (TNF‑α)] and oxidative stress indicators [superoxide dismutase (SOD), malondialdehyde (MDA) and myeloperoxidase (MPO)] were determined by ELISA. Furthermore, H9C2 cells were treated with HRS following hypoxia/reoxygenation. Cell viability and cell apoptosis were investigated. The expression of apoptosis regulator Bcl‑2 (Bcl‑2), apoptosis regulator Bax (Bax), caspase 3, AQP‑1, AQP‑4, phosphorylated (p)‑Akt, heme oxygenase 1 (HO‑1) and nuclear factor erythroid 2‑related factor 2 (Nrf2) were investigated using western blotting and quantitative‑polymerase chain reaction of tissues and cells. Following CPB, myocardial cell arrangement was disordered, myocardial injury markers (cTnI, LDH, CK‑MB and BNP), inflammatory cytokines (IL‑1β, IL‑6 and TNF‑α) and MDA levels were significantly increased compared with the sham group; whereas the SOD levels were significantly downregulated following CPB compared with the sham group. HRS attenuated myocardial injury, reduced the expression levels of cTnI, LDH, CK‑MB, BNP, IL‑1β, IL‑6, TNF‑α, MDA and MPO, and increased SOD release. Levels of Bcl‑2, AQP‑1, AQP‑4, p‑Akt, HO‑1 and Nrf2 were significantly increased following HRS; whereas Bax and caspase‑3 expression levels were significantly reduced following CPB. HRS treatment significantly increased the viability of myocardial cells, reduced the rate of myocardial cell apoptosis and the release of MDA and LDH compared with the CPB group. A PI3K inhibitor (LY294002) was revealed to reverse the protective effect of HRS treatment. HRS was demonstrated to attenuate CPB‑induced myocardial injury, suppress AQP‑1 and AQP‑4 expression following CPB treatment and protect myocardial cells via the PI3K/Akt signaling pathway.
心肌缺血、缺氧和再灌注损伤是由主动脉夹闭、心脏骤停和心肺转流术 (CPB) 期间的心脏复苏引起的,这会严重影响心脏功能。本研究旨在探讨富氢溶液 (HRS) 和水通道蛋白 (AQP) 对 CPB 诱导的心肌损伤的影响,并确定磷脂酰肌醇 3-激酶 (PI3K)/蛋白激酶 B (Akt) 信号通路的机制。将 Sprague Dawley 大鼠分为假手术组、CPB 手术组和 HRS 组。建立 CPB 模型,并在 CPB 结束时测定血流动力学参数。通过苏木精和伊红、Masson 染色观察心肌组织。通过酶联免疫吸附试验测定心肌损伤标志物 [成人心肌肌钙蛋白 I(cTnI)、乳酸脱氢酶 (LDH)、肌酸激酶 MB (CK-MB) 和脑钠肽 (BNP)]、炎症因子 [白细胞介素 (IL)-1β、IL-6 和肿瘤坏死因子-α(TNF-α)] 和氧化应激指标 [超氧化物歧化酶 (SOD)、丙二醛 (MDA) 和髓过氧化物酶 (MPO)]。此外,用 HRS 处理缺氧/复氧后的 H9C2 细胞。研究细胞活力和细胞凋亡。通过 Western blot 和组织和细胞的定量聚合酶链反应检测凋亡调节因子 Bcl-2(Bcl-2)、凋亡调节因子 Bax(Bax)、半胱天冬酶 3、AQP-1、AQP-4、磷酸化 (p)-Akt、血红素加氧酶 1 (HO-1) 和核因子红细胞 2 相关因子 2 (Nrf2) 的表达。与假手术组相比,CPB 后心肌细胞排列紊乱,心肌损伤标志物 (cTnI、LDH、CK-MB 和 BNP)、炎症细胞因子 (IL-1β、IL-6 和 TNF-α) 和 MDA 水平显著升高;而与假手术组相比,CPB 后 SOD 水平显著下调。HRS 减轻心肌损伤,降低 cTnI、LDH、CK-MB、BNP、IL-1β、IL-6、TNF-α、MDA 和 MPO 的表达水平,并增加 SOD 的释放。与 CPB 组相比,HRS 组 Bcl-2、AQP-1、AQP-4、p-Akt、HO-1 和 Nrf2 水平显著升高;而 Bax 和 caspase-3 表达水平显著降低。与 CPB 组相比,HRS 处理显著增加心肌细胞活力,降低心肌细胞凋亡率和 MDA 和 LDH 的释放。PI3K 抑制剂 (LY294002) 显示可逆转 HRS 处理的保护作用。HRS 可减轻 CPB 诱导的心肌损伤,抑制 CPB 治疗后 AQP-1 和 AQP-4 的表达,通过 PI3K/Akt 信号通路保护心肌细胞。