Li Q, Yu Q, Na R, Liu B
Zhejiang Province Hospital of Integrated Traditional Chinese and Western Medicine, Hangzhou, China.
Affiliated Zhongshan Hospital of Dalian University, Dalian, China.
Braz J Med Biol Res. 2017 May 15;50(6):e5868. doi: 10.1590/1414-431X20175868.
We aimed to investigate the effect of etanercept, a tumor necrosis factor-α (TNF-α) inhibitor, on rat cardiomyocyte hypertrophy and its underlying mechanism. Primary neonatal rat cardiomyocytes were isolated from Sprague-Dawley rats. The model of rat cardiomyocyte hypertrophy was induced by endothelin, and then treated with different concentrations of etanercept (1, 10, and 50 μM). After treatment, cell counts, viability and cell apoptosis were evaluated. The mRNA levels of myocardial hypertrophy marker genes, including atrial natriuretic factor (ANF), matrix metalloproteinase (MMP)-9 and MMP-13, were detected by qRT-PCR, and the expressions of apoptosis-related proteins (Bcl-2 and Bax) were measured by western blotting. The protein levels of transforming growth factor-β1 (TGF-β1), interleukin (IL)-1β, IL-6, leukemia inhibitory factor (LIF) and cardiotrophin-1 (CT-1) were determined using enzyme linked immunosorbent assay (ELISA) kits. In the present study, TNF-α level in cardiomyocytes with hypertrophy was significantly enhanced (P<0.05). Compared to the model group, cell number and viability were significantly increased and ratio of apoptotic cells was reduced by etanercept (P<0.05, P<0.01, or P<0.001). In addition, etanercept remarkably reduced the mRNA levels of ANF, MMP-9 and MMP-13, inhibited the expression of Bax, and increased the expression of Bcl-2 compared to the model group (P<0.05). ELISA results further showed that etanercept lowered the levels of IL-1β, IL-6, LIF and CT-1 but not TGF-β1 compared to the model group (P<0.05). Etanercept may protect rat cardiomyocytes from hypertrophy by inhibiting inflammatory cytokines secretion and cell apoptosis.
我们旨在研究肿瘤坏死因子-α（TNF-α）抑制剂依那西普对大鼠心肌细胞肥大的影响及其潜在机制。从Sprague-Dawley大鼠中分离出原代新生大鼠心肌细胞。用内皮素诱导大鼠心肌细胞肥大模型，然后用不同浓度的依那西普（1、10和50μM）进行处理。处理后，评估细胞计数、活力和细胞凋亡情况。通过qRT-PCR检测心肌肥大标志物基因心房钠尿肽（ANF）、基质金属蛋白酶（MMP）-9和MMP-13的mRNA水平，通过蛋白质印迹法检测凋亡相关蛋白（Bcl-2和Bax）的表达。使用酶联免疫吸附测定（ELISA）试剂盒测定转化生长因子-β1（TGF-β1）、白细胞介素（IL）-1β、IL-6、白血病抑制因子（LIF）和心肌营养素-1（CT-1）的蛋白水平。在本研究中，肥大心肌细胞中的TNF-α水平显著升高（P<0.05）。与模型组相比，依那西普使细胞数量和活力显著增加，凋亡细胞比例降低（P<0.05、P<0.01或P<0.001）。此外，与模型组相比，依那西普显著降低了ANF、MMP-9和MMP-13的mRNA水平，抑制了Bax的表达，并增加了Bcl-2的表达（P<0.05）。ELISA结果进一步表明，与模型组相比，依那西普降低了IL-1β、IL-6、LIF和CT-1的水平，但未降低TGF-β1的水平（P<0.05）。依那西普可能通过抑制炎性细胞因子分泌和细胞凋亡来保护大鼠心肌细胞免于肥大。
