College of Veterinary Medicine, Nanjing Agricultural University, Nanjing 210095, PR China.
College of Animal Science and Technology, Nanjing Agricultural University, Nanjing 210095, PR China.
Int Immunopharmacol. 2020 Jun;83:106392. doi: 10.1016/j.intimp.2020.106392. Epub 2020 Mar 14.
The anti-inflammatory effects of sodium valproate (VPA) in vivo and in vitro have been demonstrated in recent studies. The aim of this study was to evaluate whether VPA can suppress inflammation in bovine mammary epithelial cells (BMECs) stimulated by γ-D-glutamyl-meso-diaminopimelic acid (iE-DAP). First, the concentration and treatment points of iE-DAP and VPA were optimized. Then, BMECs were cultured in complete media and separated into four groups: untreated control cells (CON group), cells stimulated by 10 μg/mL iE-DAP for 6 h (DAP group), cells stimulated by 0.5 mmol/L VPA for 6 h (VPA group), and cells pretreated with VPA (0.5 mmol/L) for 6 h followed by 10 μg/mL of iE-DAP for 6 h (VD group). The results showed that the level of interleukin-6 (IL-6) and tumor necrosis factor-α (TNF-α) in the culture medium increased in the iE-DAP-treated cells and that pretreatment with VPA reversed this increase. iE-DAP increased both mRNA and protein expression levels of nucleotide-binding oligomerization domain-containing protein 1 (NOD1) and receptor-interacting protein kinas (RIPK2) and activated inhibitor of NF-κB (IκB) and nuclear factor-kappa B p65 (NF-κB p65) through phosphorylation. Upon activation of the NF-κB pathway, the expression of the pro-inflammatory cytokines IL-6, interleukin-8 (IL-8) and interleukin-1β (IL-1β), the acute phase protein serum amyloid A 3 (SAA3) and the lingual antimicrobial peptide (LAP) but not haptoglobi (HP) or bovine neutrophil beta defensing 5 (BNBD5) were increased in the DAP group. The VPA pretreatment induced the acetylation of signal transducers and activators of transcription(STAT1) and histone 3 (H3) by inhibiting histone deacetylase (HDAC) and then suppressed the NF-κB pathway. Moreover, VPA induced autophagy and reduced apoptosis in BMECs in the VD group. These results suggested that VPA treatment can attenuate the inflammatory response induced by iE-DAP.
最近的研究表明，丙戊酸钠（VPA）具有体内和体外的抗炎作用。本研究旨在评估 VPA 是否能抑制γ-D-谷氨酰基-中二氨基庚二酸（iE-DAP）刺激的牛乳腺上皮细胞（BMECs）中的炎症。首先，优化了 iE-DAP 和 VPA 的浓度和处理点。然后，将 BMECs 在完全培养基中培养，并分为四组：未处理对照细胞（CON 组）、用 10μg/mL iE-DAP 刺激 6 小时的细胞（DAP 组）、用 0.5mmol/L VPA 刺激 6 小时的细胞（VPA 组）和用 VPA（0.5mmol/L）预处理 6 小时后用 10μg/mL iE-DAP 刺激 6 小时的细胞（VD 组）。结果表明，iE-DAP 处理的细胞培养基中白细胞介素-6（IL-6）和肿瘤坏死因子-α（TNF-α）水平升高，而 VPA 预处理则逆转了这种升高。iE-DAP 增加了核苷酸结合寡聚化结构域蛋白 1（NOD1）和受体相互作用蛋白激酶（RIPK2）的 mRNA 和蛋白表达水平，并通过磷酸化激活了核因子-κB 抑制蛋白（IκB）和核因子-κB p65（NF-κB p65）。在 NF-κB 途径被激活后，促炎细胞因子 IL-6、白细胞介素-8（IL-8）和白细胞介素-1β（IL-1β）、急性期蛋白血清淀粉样蛋白 A3（SAA3）和舌抗菌肽（LAP）的表达增加，但触珠蛋白（HP）或牛中性粒细胞β防御素 5（BNBD5）的表达没有增加，在 DAP 组中。VPA 预处理通过抑制组蛋白去乙酰化酶（HDAC）诱导信号转导和转录激活因子（STAT1）和组蛋白 3（H3）的乙酰化，从而抑制 NF-κB 途径。此外，VPA 在 VD 组中诱导了自噬并减少了 BMECs 的凋亡。这些结果表明，VPA 治疗可以减轻 iE-DAP 诱导的炎症反应。
