Yu T, Dong D, Guan J, Sun J, Guo M, Wang Q
Department of Emergency, Affiliated Hospital of Qingdao University, Jiangsu Road No. 16, Qingdao, Shandong, China.
Department of Cardiology, No. 971 Hospital of People's Liberation Army, Minjiang Road No. 22, Qingdao, Shandong, China.
Herz. 2020 Dec;45(Suppl 1):130-138. doi: 10.1007/s00059-019-4837-0. Epub 2019 Jul 16.
Clinical research has demonstrated that alprostadil has an anti-inflammatory effect; however, to date, its molecular mechanisms remain unclear. This study aimed to examine the anti-inflammatory activity and related mechanisms of alprostadil in lipopolysaccharide (LPS)-treated H9c2 cells.
Cell morphology was observed under an inverted light microscope, while cell viability was assessed with the 3‑(4,5-dimethylthiazolyl-2)-2,5-diphenyltetrazolium bromide (MTT) assay. Enzyme-linked immunosorbent assays (ELISA) were conducted to study biochemical indicators of cellular damage, such as released lactate dehydrase (LDH) and troponin, and inflammatory cytokine levels including interleukin-1β (IL-1β), IL-6, IL-17, and tumor necrosis factor-α (TNF-α). The mRNA expression levels of Wnt5a, c‑jun N‑terminal kinase (JNK), and nuclear factor kappa B (NF-κB) were further investigated by real-time quantitative polymerase chain reaction (RT-PCR). The effects of alprostadil on the Wnt5a/JNK/NF-κB pathway in H9c2 cells was examined by Western blotting.
Alprostadil increased the cell viability of LPS-stimulated H9c2 cells, reduced LDH and troponin production, and attenuated IL-1β, IL-6, IL-17, and TNF-α secretion. Moreover, alprostadil reduced the mRNA expression of Wnt5a, JNK, and NF-κB and decreased the expression of Wnt5a, NF-κB, and the ratio of p‑JNK/JNK in H9c2 cells treated with LPS. The siWnt5a or JNK inhibitor SP600125 significantly augmented the inhibitory effects of alprostadil on the Wnt5a/JNK/NF-κB pathway.
Our results show that alprostadil has anti-inflammatory effects and could attenuate LPS-induced injury in H9c2 cardiomyocytes via the Wnt5a/JNK/NF-κB pathway.
临床研究表明，前列地尔具有抗炎作用；然而，迄今为止，其分子机制仍不清楚。本研究旨在探讨前列地尔在脂多糖（LPS）处理的H9c2细胞中的抗炎活性及相关机制。
在倒置光学显微镜下观察细胞形态，同时用3-(4,5-二甲基噻唑-2)-2,5-二苯基四氮唑溴盐（MTT）法评估细胞活力。进行酶联免疫吸附测定（ELISA）以研究细胞损伤的生化指标，如释放的乳酸脱氢酶（LDH）和肌钙蛋白，以及炎症细胞因子水平，包括白细胞介素-1β（IL-1β）、IL-6、IL-17和肿瘤坏死因子-α（TNF-α）。通过实时定量聚合酶链反应（RT-PCR）进一步研究Wnt5a、c-Jun氨基末端激酶（JNK）和核因子κB（NF-κB）的mRNA表达水平。通过蛋白质印迹法检测前列地尔对H9c2细胞中Wnt5a/JNK/NF-κB信号通路的影响。
前列地尔提高了LPS刺激的H9c2细胞的活力，降低了LDH和肌钙蛋白的产生，并减弱了IL-1β、IL-6、IL-17和TNF-α的分泌。此外，前列地尔降低了LPS处理的H9c2细胞中Wnt5a、JNK和NF-κB的mRNA表达，并降低了Wnt5a、NF-κB的表达以及p-JNK/JNK的比值。siWnt5a或JNK抑制剂SP600125显著增强了前列地尔对Wnt5a/JNK/NF-κB信号通路的抑制作用。
我们的结果表明，前列地尔具有抗炎作用，并可通过Wnt5a/JNK/NF-κB信号通路减轻LPS诱导的H9c2心肌细胞损伤。
