Antioxidant and Antineuroinflammatory Mechanisms of Kaempferol-3- O-β-d-Glucuronate on Lipopolysaccharide-Stimulated BV2 Microglial Cells through the Nrf2/HO-1 Signaling Cascade and MAPK/NF-κB Pathway

Hyun Jung Lim; Ritu Prajapati; Su Hui Seong; Hyun Ah Jung; Jae Sue Choi

doi:10.1021/acsomega.2c06916

Antioxidant and Antineuroinflammatory Mechanisms of Kaempferol-3- O-β-d-Glucuronate on Lipopolysaccharide-Stimulated BV2 Microglial Cells through the Nrf2/HO-1 Signaling Cascade and MAPK/NF-κB Pathway

ACS Omega. 2023 Feb 10;8(7):6538-6549. doi: 10.1021/acsomega.2c06916. eCollection 2023 Feb 21.

Authors

Hyun Jung Lim¹, Ritu Prajapati², Su Hui Seong³, Hyun Ah Jung⁴, Jae Sue Choi^{1

2}

Affiliations

¹ Institute of Fisheries Sciences, Pukyong National University, Busan 46041, Republic of Korea.
² Department of Food and Life Science, Pukyong National University, Busan 48513, Republic of Korea.
³ Division of Natural Products Research, Honam National Institute of Biological Resource, Mokpo 58762, Republic of Korea.
⁴ Department of Food Science and Human Nutrition, Jeonbuk National University, Jeonju 54896, Republic of Korea.

Abstract

Aglycone- and glycoside-derived forms of flavonoids exist broadly in plants and foods such as fruits, vegetables, and peanuts. However, most studies focus on the bioavailability of flavonoid aglycone rather than its glycosylated form. Kaempferol-3-O-β-d-glucuronate (K3G) is a natural flavonoid glycoside obtained from various plants that have several biological activities, including antioxidant and anti-inflammatory effects. However, the molecular mechanism related to the antioxidant and antineuroinflammatory activity of K3G has not yet been demonstrated. The present study was designed to demonstrate the antioxidant and antineuroinflammatory effect of K3G against lipopolysaccharide (LPS)-stimulated BV2 microglial cells and to evaluate the underlying mechanism. Cell viability was determined by MTT assay. The inhibition rate of reactive oxygen species (ROS) and the production of pro-inflammatory mediators and cytokines were measured by DCF-DA assay, Griess assay, enzyme-linked immunosorbent assay (ELISA), and western blotting. K3G inhibited the LPS-induced release of nitric oxide, interleukin (IL)-6, and tumor necrosis factor-α (TNF-α) as well as the expression of prostaglandin E synthase 2. Additionally, K3G reduced the expression of inducible nitric oxide synthase (iNOS), cyclooxygenase-2 (COX-2), and nuclear factor-kappa B (NF-κB) related proteins. Mechanistic studies found that K3G downregulated phosphorylated mitogen-activated protein kinases (MAPKs) and upregulated the Nrf2/HO-1 signaling cascade. In this study, we demonstrated the effects of K3G on antineuroinflammation by inactivating phosphorylation of MPAKs and on antioxidants by upregulating the Nrf2/HO-1 signaling pathway through decreasing ROS in LPS-stimulated BV2 cells.