The Effects of Aronia melanocarpa ‘Viking’ Extracts in Attenuating RANKL-Induced Osteoclastic Differentiation by Inhibiting ROS Generation and c-FOS/NFATc1 Signaling

Abstract

This study aimed to determine the anti-osteoclastogenic effects of extracts from Aronia melanocarpa ‘Viking’ (AM) and identify the underlying mechanisms in vitro. Reactive oxygen species (ROS) are signal mediators in osteoclast differentiation. AM extracts inhibited ROS production in RAW 264.7 cells in a dose-dependent manner and exhibited strong radical scavenging activity. The extracts also attenuated the number of tartrate-resistant acid phosphatase (TRAP)-positive multinucleated osteoclasts. To attain molecular insights, the effect of the extracts on the signaling pathways induced by receptor activator of nuclear factor kappa B ligand (RANKL) were also investigated. RANKL triggers many transcription factors through the activation of mitogen-activated protein kinase (MAPK) and ROS, leading to the induction of osteoclast-specific genes. The extracts significantly suppressed RANKL-induced activation of MAPKs, such as extracellular signal-regulated kinase (ERK), c-Jun-N-terminal kinase (JNK) and p38 and consequently led to the downregulation of c-Fos and nuclear factor of activated T cells 1 (NFATc1) protein expression which ultimately suppress the activation of the osteoclast-specific genes, cathepsin K, TRAP, calcitonin receptor and integrin β3. In conclusion, our findings suggest that AM extracts inhibited RANKL-induced osteoclast differentiation by downregulating ROS generation and inactivating JNK/ERK/p38, nuclear factor kappa B (NF-κB)-mediated c-Fos and NFATc1 signaling pathway.

6,7,4′‐Trihydroxyflavone inhibits osteoclast formation and bone resorption in vitro and in vivo

“…Activation of c‐Fos leads to the expression of NFATc1, an endogenous regulator of osteoclast (Izawa et al, ). Also, RANKL induces many osteoclast‐specific genes such as cathepsin K and c‐src through activation of various signaling pathways, thereby increasing osteoclast activity and bone resorption (Ghosh et al, ; Izawa et al, ). Therefore, the effect of THF on the expression of c‐Fos, NFATc1, cathepsin K and c‐src in RANKL‐induced RAW 264.7 (Figure a) and BMM cells (Figure b) was evaluated by Western blotting.…”

“…Activation of c-Fos leads to the expression of NFATc1, an endogenous regulator of osteoclast (Izawa et al, 2016). Also, RANKL induces many osteoclast-specific genes such as cathepsin K and c-src through activation of various signaling pathways, thereby increasing osteoclast activity and bone resorption (Ghosh et al, 2018;Izawa et al, 2016).…”

Green chicory leaf extract exerts anti-inflammatory effects through suppressing LPS-induced MAPK/NF-κB activation and hepatoprotective activity in vitro

“…Besides, chicory root is widely used as functional food components as well in various ways (Sinkovič et al, 2015). Natural functional foods/components previously showed remarkable effects against inflammatory diseases (Ghosh et al, 2018;Hasnat, Pervin, Cha, Kim, & Lim, 2015;Miao et al, 2019;Pervin et al, 2016). Chicory leaves are cost-effective with many health benefits and may consume raw in salad and there are many varieties of chicory are available.…”

“…Inflammation is an inevitable and essential cellular event in response to infection, antigen in the living organisms. The inflammation is also recognized as a hallmark of many chronic diseases, such as cancer, obesity, diabetes, arthritis, cardiac diseases, IBD, Parkinson disease (PD) and so on (Al Mijan & Lim, 2018;Chen et al, 2018;Ghosh et al, 2018;Miao et al, 2019;Tilg, Zmora, Adolph, & Elinav, 2020) when it goes aversively. Molecular mechanisms of inflammation are still complex and not fully understood.…”

“…macrophages, neutrophils, B cells and so on). RAW 264.7 cell line (murine macrophage) is well established and extensively used for anti-inflammatory studies (Ghosh et al, 2018;Hasnat et al, 2015;Miao et al, 2019;Ng, Zainal Abidin, Shuib, & Israf Ali, 2015). Splenocytes contain B cells, T cells, Monocytes and other immune cells that protect the host from foreign invaders and pivotal inflammatory mediators as well.…”