Department of Emergency, Affiliated Hospital of Jining Medical University, Jining, 272000, Shandong, China.
Department of Cardiac Surgery, Jining No. 1 People's Hospital, No. 6, Jiankang Road, Jining, 272000, Shandong, China.
Inflammation. 2018 Aug;41(4):1229-1237. doi: 10.1007/s10753-018-0769-8.
Myocarditis is a cardiomyopathy associated with inflammatory response. It has been reported that geniposide (GEN), a traditional Chinese herb extract from Gardenia jasminoides Ellis, possesses an anti-inflammatory effect and a protective effect on cardiomyocytes. The present study aimed to explore the protective role of GEN and the underlying mechanism in LPS-injured H9c2 cells. H9c2 cells were treated with LPS to induce cell injury and then we investigated the effect of GEN. miR-145 expression was inhibited by transfection with miR-145 inhibitor and its expression was measured by RT-PCR. Cell viability and apoptotic cells were measured by CCK-8 assay and flow cytometry analysis. The levels of pro-inflammatory factors (IL-6, TNF-α, and MCP-1) were assessed by western blot and RT-PCR. Western blot was performed to detect the expression of the MEK/ERK pathway-related factors. LPS exposure reduced cell viability, increased apoptotic cells, and promoted the expression of pro-inflammatory factors in H9c2 cells. However, GEN pretreatment significantly reduced LPS-induced cell injury, as increased cell viability, reduced apoptotic cells, and inhibited the expression of pro-inflammatory factors. Moreover, we found that miR-145 expression was down-regulated by LPS exposure but was up-regulated by GEN pretreatment. The protective effect of GEN on LPS-injured H9c2 cells was blocked by miR-145 inhibitor. In addition, GEN inhibited the MEK/ERK pathway through up-regulating miR-145. Our results suggested that GEN exerted a protective role in LPS-injured H9c2 cells. The GEN-associated regulation might be related to its regulation on miR-145 and the MEK/ERK signaling pathway.
心肌炎是一种与炎症反应相关的心肌病。有报道称,栀子苷(GEN)是从栀子(Gardenia jasminoides Ellis)中提取的一种中药提取物,具有抗炎作用和对心肌细胞的保护作用。本研究旨在探讨 GEN 在 LPS 损伤的 H9c2 细胞中的保护作用及其潜在机制。用 LPS 处理 H9c2 细胞以诱导细胞损伤,然后我们研究了 GEN 的作用。通过转染 miR-145 抑制剂抑制 miR-145 的表达,并通过 RT-PCR 测量其表达。通过 CCK-8 测定法和流式细胞术分析测量细胞活力和凋亡细胞。通过 Western blot 和 RT-PCR 评估促炎因子(IL-6、TNF-α 和 MCP-1)的水平。Western blot 用于检测 MEK/ERK 通路相关因子的表达。LPS 暴露降低细胞活力,增加凋亡细胞,并促进 H9c2 细胞中促炎因子的表达。然而,GEN 预处理可显著减轻 LPS 诱导的细胞损伤,增加细胞活力,减少凋亡细胞,并抑制促炎因子的表达。此外,我们发现 LPS 暴露下调 miR-145 的表达,但 GEN 预处理可上调 miR-145 的表达。GEN 对 LPS 损伤的 H9c2 细胞的保护作用被 miR-145 抑制剂阻断。此外,GEN 通过上调 miR-145 抑制 MEK/ERK 通路。我们的研究结果表明,GEN 对 LPS 损伤的 H9c2 细胞发挥了保护作用。GEN 相关的调节可能与其对 miR-145 和 MEK/ERK 信号通路的调节有关。