College of Veterinary Medicine, Jeju National University, Jeju 63243, Korea.
J Vet Sci. 2020 Nov;21(6):e91. doi: 10.4142/jvs.2020.21.e91.
Sulforaphane (SFN) is an isothiocyanate compound present in cruciferous vegetables. Although the anti-inflammatory effects of SFN have been reported, the precise mechanism related to the inflammatory genes is poorly understood.
This study examined the relationship between the anti-inflammatory effects of SFN and the differential gene expression pattern in SFN treated ob/ob mice.
Nitric oxide (NO) level was measured using a Griess assay. The inducible nitric oxide synthase (iNOS) and cyclooxygenase-2 (COX-2) expression levels were analyzed by Western blot analysis. Pro-inflammatory cytokines (tumor necrosis factor [TNF]-α, interleukin [IL]-1β, and IL-6) were measured by enzyme-linked immunosorbent assay (ELISA). RNA sequencing analysis was performed to evaluate the differential gene expression in the liver of ob/ob mice.
The SFN treatment significantly attenuated the iNOS and COX-2 expression levels and inhibited NO, TNF-α, IL-1β, and IL-6 production in lipopolysaccharide (LPS)-stimulated RAW 264.7 cells. RNA sequencing analysis showed that the expression levels of 28 genes related to inflammation were up-regulated (> 2-fold), and six genes were down-regulated (< 0.6-fold) in the control ob/ob mice compared to normal mice. In contrast, the gene expression levels were restored to the normal level by SFN. The protein-protein interaction (PPI) network showed that chemokine ligand (Cxcl14, Ccl1, Ccl3, Ccl4, Ccl17) and chemokine receptor (Ccr3, Cxcr1, Ccr10) were located in close proximity and formed a "functional cluster" in the middle of the network.
The overall results suggest that SFN has a potent anti-inflammatory effect by normalizing the expression levels of the genes related to inflammation that were perturbed in ob/ob mice.
萝卜硫素（SFN）是十字花科蔬菜中存在的一种异硫氰酸酯化合物。尽管 SFN 的抗炎作用已被报道，但与炎症基因相关的确切机制尚不清楚。
本研究旨在探讨 SFN 的抗炎作用与 SFN 处理 ob/ob 小鼠中差异基因表达模式之间的关系。
通过格里斯测定法测量一氧化氮（NO）水平。通过 Western blot 分析分析诱导型一氧化氮合酶（iNOS）和环氧化酶-2（COX-2）的表达水平。通过酶联免疫吸附测定（ELISA）测量促炎细胞因子（肿瘤坏死因子[TNF]-α、白细胞介素[IL]-1β和 IL-6）。进行 RNA 测序分析以评估 ob/ob 小鼠肝脏中的差异基因表达。
SFN 处理显著减弱了 LPS 刺激的 RAW 264.7 细胞中 iNOS 和 COX-2 的表达水平，并抑制了 NO、TNF-α、IL-1β 和 IL-6 的产生。RNA 测序分析显示，与正常小鼠相比，对照 ob/ob 小鼠中 28 个与炎症相关的基因表达水平上调（>2 倍），6 个基因下调（<0.6 倍）。相比之下，SFN 将基因表达水平恢复到正常水平。蛋白质-蛋白质相互作用（PPI）网络显示趋化因子配体（Cxcl14、Ccl1、Ccl3、Ccl4、Ccl17）和趋化因子受体（Ccr3、Cxcr1、Ccr10）彼此靠近，并且在网络的中间形成了一个“功能簇”。
总体结果表明，SFN 通过使 ob/ob 小鼠中失调的与炎症相关的基因表达水平正常化，具有强大的抗炎作用。
