The First Department of Cadres Health Care, The Third Hospital of Shijiazhuang, Shijiazhuang, Hebei 050011, P.R. China.
The Second Department of Cadres Health Care, The Third Hospital of Shijiazhuang, Shijiazhuang, Hebei 050011, P.R. China.
Mol Med Rep. 2019 May;19(5):4109-4118. doi: 10.3892/mmr.2019.10077. Epub 2019 Mar 22.
The present study aimed to investigate the potential effects of growth differentiation factor 11 (GDF11) on isoproterenol (ISO)‑induced heart failure (HF) and identify the underlying molecular mechanisms. A rat model of HF was induced in vivo by intraperitoneally administering ISO (5 mg/kg/day) for 7 days. After 4 weeks following establishment of the HF model, hemodynamic analysis demonstrated that ISO induced a significant increase in the left ventricular end‑diastolic pressure and a decrease in the left ventricular systolic pressure and maximum contraction velocity. The plasma levels of myocardial injury markers, including lactate dehydrogenase (LDH), creatine kinase (CK), CK‑muscle/brain which were determined using the corresponding assay kits and plasma brain natriuretic peptide which was detected by an ELISA kit, an important biomarker of HF, increased following ISO treatment. Furthermore, levels of GDF11 expression and protein, which were estimated using reverse transcription‑quantitative polymerase chain reaction and an ELISA kit in plasma and western blotting in the heart tissue, respectively, significantly increased following ISO treatment. To demonstrate the effects of ISO on GDF11 production in cardiomyocytes, H9C2 cells (a cardiomyoblast cell line derived from embryonic rat heart tissue) were treated with ISO (50 nM) for 24 h in vitro; it was revealed that GDF11 protein and mRNA expression levels significantly increased following ISO treatment. In addition, recombinant GDF11 (rGDF11) administered to ISO‑treated H9C2 cells resulted in decreased proliferation, which was detected via a CCK‑8 assay, and increased LDH levels and cell apoptosis of cells, which was determined using Caspase‑3 activity and Hoechst 33258 staining. Additionally, rGDF11 increased the levels of reactive oxygen species and malondialdehyde due to the upregulation of nicotinamide adenine dinucleotide phosphate oxidase 4 (Nox4) following rGDF11 treatment. Conversely, GDF11 knockdown reduced ISO‑induced apoptosis by inhibiting oxidative stress injury. The results suggested that GDF11 production was upregulated in ISO‑induced rats with HF and in ISO‑treated H9C2 cells, and that rGDF11 treatment increased ISO‑induced oxidative stress injury by upregulating Nox4 in H9C2 cells.
本研究旨在探讨生长分化因子 11（GDF11）对异丙肾上腺素（ISO）诱导的心力衰竭（HF）的潜在影响，并确定其潜在的分子机制。通过腹腔内给予 ISO（5mg/kg/天）7 天在体内诱导 HF 大鼠模型。HF 模型建立 4 周后，血流动力学分析表明 ISO 导致左心室舒张末期压力显著增加，左心室收缩压和最大收缩速度降低。通过相应的试剂盒测定血浆心肌损伤标志物，包括乳酸脱氢酶（LDH）、肌酸激酶（CK）、肌/脑型 CK 以及通过 ELISA 试剂盒检测的脑钠肽（BNP），HF 的重要生物标志物，ISO 处理后升高。此外，使用逆转录定量聚合酶链反应和 ELISA 试剂盒分别在血浆和心脏组织中Western 印迹法检测到 GDF11 表达和蛋白水平在 ISO 处理后显著增加。为了证明 ISO 对心肌细胞中 GDF11 产生的影响，将 H9C2 细胞（源自胚胎大鼠心脏组织的心肌细胞系）在体外用 ISO（50nM）处理 24 小时；结果表明，ISO 处理后 GDF11 蛋白和 mRNA 表达水平显著增加。此外，重组 GDF11（rGDF11）给予 ISO 处理的 H9C2 细胞导致增殖减少，通过 CCK-8 测定检测到，以及细胞 LDH 水平和细胞凋亡增加，通过 Caspase-3 活性和 Hoechst 33258 染色测定。此外，rGDF11 增加了活性氧和丙二醛的水平，因为 rGDF11 处理后烟酰胺腺嘌呤二核苷酸磷酸氧化酶 4（Nox4）上调。相反，GDF11 敲低通过抑制氧化应激损伤减少 ISO 诱导的细胞凋亡。结果表明，在 ISO 诱导的 HF 大鼠和 ISO 处理的 H9C2 细胞中 GDF11 的产生上调，rGDF11 处理通过上调 H9C2 细胞中的 Nox4 增加 ISO 诱导的氧化应激损伤。
