Phenylethanoid Glycosides From Callicarpa kwangtungensis Chun Attenuate TNF-α-Induced Cell Damage by Inhibiting NF-κB Pathway and Enhancing Nrf2 Pathway in A549 Cells

Abstract

Background: Acute lung injury (ALI) is a complicated and severe lung disease, which is often characterized by acute inflammation. Poliumoside (POL), acteoside (ACT) and forsythiaside B (FTB) are phenylethanoid glycosides (PGs) with strong antioxidant, anti-inflammatory, and anti-apoptotic properties, which are extracted from Callicarpa kwangtungensis Chun (CK). The aim of this study was to investigate the protective effects of POL, ACT, and FTB against TNF-α-induced damage using an ALI cell model and explore their potential mechanisms.Methods and Results: MTT method was used to measure cell viability. Flow cytometry was used for detecting the apoptosis rate. Reactive oxygen species (ROS) activity was determined using fluorescence microscope. The expression of mRNA in apoptosis-related genes (Caspase 3, Caspase 8, and Caspase 9) were tested by qPCR. The effects of POL, ACT, FTB on the activities of nuclear factor erythroid-2 related factor 2 (Nrf2), nuclear factor kappa-B (NF-κB) and the expression of their downstream genes were assessed by western blotting and RT-PCR in A549 cells. In the current study, POL, ACT, and FTB dose-dependently attenuated TNF-α-induced IL-1β, IL-6 and IL-8 production, cell apoptosis, the expression of apoptosis-related genes (Caspase 3, Caspase 8, and Caspase 9) and ROS activity. POL, ACT, and FTB not only increased in the mRNA levels of antioxidative enzymes NADPH quinone oxidoreductase (NQO1), glutamate cysteine ligase catalytic subunit (GCLC), heme oxygenase (HO-1), but also decreased the mRNA levels of IL-1β, IL-6 and IL-8. Furthermore, they upregulated the expression of Keap1 and enhanced the activation of Nrf2, while decreased the expression of phosphor-IκBα (p-IκBα) and nuclear p65. In addition, no significant changes were observed in anti-inflammatory and antioxidant effects of POL, ACT, FTB following Nrf2 and NF-κB p65 knockdown.Conclusion: Our study revealed that POL, ACT, and FTB alleviated oxidative damage and lung inflammation of TNF-α-induced ALI cell model through regulating the Nrf2 and NF-κB pathways.

The pharmacokinetic property and pharmacological activity of acteoside: A review

“… [40] A549 Decreases NF-κB, IL-1β, IL-6, IL-8, and caspase-3/-8/-9, increases NRF2, HO-1, NQO-1, and GCLC. [42] U937 Induces SHP-1 phosphorylation, attenuates the activation of TAK1/JNK/AP-1, and decreases COX and NOS expression. [44] Dendritic cells Increases IL-10 expression by activating AhR expression.…”

“…AC has been shown to dose-dependently ameliorate TNFα-induced IL-1β, IL-6, IL-8, and caspase-3/-8/-9 expression, ROS generation, and cell apoptosis. In addition, AC increases the expression of NRF2, HO-1, NADPH quinone oxidoreductase (NQO-1), and glutamate cysteine ligase catalytic subunit (GCLC) and decreases the activity of NF-κB signaling pathway in TNFα-treated A549 cells [42] . In lipopolysaccharide (LPS)-induced acute lung injury, AC exhibits protective activity against the histopathological changes, the increase of proinflammatory cytokines, and the enhancement of NF-κB signaling pathway [43] .…”

Glycyl-L-histidyl-L-lysine-Cu2+ attenuates cigarette smoke-induced pulmonary emphysema and inflammation by reducing oxidative stress pathway

“…However, under unusually severe oxidative stress, the Keap1-Nrf2 mechanism becomes disrupted and results in cell and tissue damage. These paradoxical results could be observed in some other studies, such as the Keap1-Nrf2 axis in acute lung injury, in which Keap1 expression increased in some animal and in vitro experiments (Huang et al, 2020;Li et al, 2021), while Keap1 expression decreased in some animal and in vitro experiments (Yan et al, 2019;Zheng et al, 2021;Zhu et al, 2021). MMPs and TIMPs imbalance plays a key role in the destruction of lung parenchyma (Mocchegiani et al, 2011).…”

Antioxidant and Anti-Inflammatory Activity of Combined Phycocyanin and Palmitoylethanolamide in Human Lung and Prostate Epithelial Cells

“…Indeed, in Beas-2B cells, the incubation with PC, PEA and the inflammatory stimulus seemed to have an addictive effect, possibly mediated by the induction of the endogenous synthesis of glutathione. To verify this assumption, we measured the transcription of a key enzyme in GSH biosynthesis, namely the catalytic subunit of the glutamate-cysteine ligase (GCLC) enzyme [ 37 ]. We discovered that both in Beas-2B ( Figure 3 B) and prostate HPrEpiC ( Figure 3 C) the two molecules acted on gene transcription of the enzyme that drives GSH production.…”