Sodium valproate attenuates the iE-DAP induced inflammatory response by inhibiting the NOD1-NF-κB pathway and histone modifications in bovine mammary epithelial cells
Sodium valproate attenuates the iE-DAP induced inflammatory response by inhibiting the NOD1-NF-κB pathway and histone modifications in bovine mammary epithelial cells
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丙戊酸钠通过抑制牛乳腺上皮细胞中的 NOD1-NF-kappa B 通路和组蛋白修饰来减轻 iE-DAP 诱导的炎症反应
DOI：
10.1016/j.intimp.2020.106392
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发表时间：
2020-06-01
影响因子：
5.6
通讯作者：
Shen, Xiangzhen
中科院分区：
文献类型：
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作者：
Gao, Qianyun;Wang, Yan;Shen, Xiangzhen
近年来的研究表明，丙戊酸钠(VPA)在体内和体外均有抗炎作用。本研究旨在探讨丙戊酸(VPA)能否抑制γ-D-谷氨酰-中位二氨基戊二酸(IE-DAP)刺激的牛乳腺上皮细胞(BMECs)炎症反应。首先，对IE-DAP和VPA的浓度和处理点进行了优化。将体外培养的BMEC分为4组：空白对照组(CON组)、10mU/mLIe-DAP刺激6h组(DAP组)、0.5 mmol/L VPA刺激6h组(VPA组)、VPA(0.5 mmoL/L)处理6h+10mug/mLie-DAP刺激6h组(VD组)。结果表明，经IE-DAP处理的细胞培养上清液中IL-6和肿瘤坏死因子-α水平升高，而VPA可逆转这一升高。IE-DAP通过磷酸化增加核苷酸结合的寡聚化结构域含蛋白1(NOD1)、受体相互作用蛋白Kinas(RIPK2)、活化的核因子-kappaB抑制物(I-kappa B)和核因子-kappaBp65(NF-kappa Bp65)的mRNA和蛋白表达水平。激活核因子-kappaB通路后，DAP组促炎细胞因子IL-6、IL-8和IL-1β、急性时相蛋白血清淀粉样蛋白A3(SAA3)和舌抗菌肽(LAP)的表达增加，而触珠蛋白(HP)和牛中性粒细胞β防御因子5(BNBD5)的表达不增加。VPA可通过抑制组蛋白脱乙酰酶(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，进而抑制NF-kappa B通路。此外，VPA可诱导VD组BMECs自噬，减少细胞凋亡。提示VPA治疗可减轻IE-DAP诱导的炎症反应。
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 gamma-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 mu g/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 mu g/mL of iE-DAP for 6 h (VD group). The results showed that the level of interleukin-6 (IL-6) and tumor necrosis factor-alpha (TNF-alpha) 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-kappa B (I kappa B) and nuclear factor-kappa B p65 (NF-kappa B p65) through phosphorylation. Upon activation of the NF-kappa B pathway, the expression of the pro-inflammatory cytokines IL-6, interleukin-8 (IL-8) and interleukin-1 beta (IL-1 beta), 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-kappa B 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.