Department of Infectious Diseases, Renmin Hospital of Wuhan University, Wuhan, Hubei 430060, China.
Department of Infectious Diseases, Renmin Hospital of Wuhan University, Wuhan, Hubei 430060, China.
Biomed Pharmacother. 2019 Sep;117:109166. doi: 10.1016/j.biopha.2019.109166. Epub 2019 Jun 27.
Histone deacetylase 6 (HDAC6) has been considered as an important regulator in the development of inflammatory diseases. However, the mechanism of HDAC6 in regulating inflammatory responses has not been fully determined. In the present study, we aim to investigate the role and mechanisms of HDAC6 in regulating inflammation in lipopolysaccharide (LPS)-activated macrophages.
Flow cytometry was used to determine a suitable treatment dosage of ACY-1215 on lipopolysaccharide (LPS)-activated macrophages for the present study. The RAW264.7 macrophages were divided into normal, LPS-treated, and ACY-1215 treated groups, respectively. For the ACY-1215 group, ACY-1215 (10 μM) was added to the medium 2 h prior to treatment with LPS (1 μg/ml) for 24 h. In this study, ROS, inflammatory cytokines, the ultrastructure of mitochondria, mitochondrial membrane potential, RNA and protein expression assay were detected respectively. Subsequently, the effect of HDAC6 knockdown on inflammatory response in LPS-activated RAW264.7 macrophages was also detected.
Inhibition of HDAC6 inhibited the overproduction of ROS and suppressed the expression of pro-inflammatory cytokines such as TNF-α, IL-1β, and IL-6 in LPS-activated RAW264.7 cells. Pretreatment with ACY-1215 could normalize the ultrastructure of mitochondria and mitochondrial membrane potential in LPS-activated macrophages. Moreover, the protein expression of TLR4, Nrf2, HO-1 and the activation of MAPK and NF-κB signaling pathways were normalized by the inhibition of HDAC6.
Inhibition of HDAC6 exhibited protective role against LPS-induced inflammation in RAW264.7 cells by regulating oxidative stress and suppressing the activation of TLR4- MAPK/NF-κB signaling pathway.
组蛋白去乙酰化酶 6（HDAC6）被认为是炎症性疾病发展的重要调节因子。然而，HDAC6 调节炎症反应的机制尚未完全确定。在本研究中，我们旨在研究 HDAC6 在调节脂多糖（LPS）激活的巨噬细胞炎症中的作用和机制。
流式细胞术用于确定本研究中 ACY-1215 对脂多糖（LPS）激活的巨噬细胞的合适处理剂量。将 RAW264.7 巨噬细胞分为正常组、LPS 处理组和 ACY-1215 处理组，分别进行处理。对于 ACY-1215 组，在 LPS（1μg/ml）处理 24 小时前，将 ACY-1215（10μM）加入培养基中 2 小时。在本研究中，分别检测 ROS、炎症细胞因子、线粒体超微结构、线粒体膜电位、RNA 和蛋白质表达测定。随后，还检测了 HDAC6 敲低对 LPS 激活的 RAW264.7 巨噬细胞炎症反应的影响。
抑制 HDAC6 抑制了 ROS 的过度产生，并抑制了 LPS 激活的 RAW264.7 细胞中 TNF-α、IL-1β 和 IL-6 等促炎细胞因子的表达。ACY-1215 的预处理可以使 LPS 激活的巨噬细胞中线粒体的超微结构和线粒体膜电位正常化。此外，HDAC6 的抑制还使 TLR4、Nrf2、HO-1 的蛋白表达以及 MAPK 和 NF-κB 信号通路的激活正常化。
抑制 HDAC6 通过调节氧化应激和抑制 TLR4-MAPK/NF-κB 信号通路的激活，对 LPS 诱导的 RAW264.7 细胞炎症发挥保护作用。
