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Nobiletin ameliorates hormone-induced osteoblast apoptosis by modulating JAK2/STAT3 signaling
Xiang Li1, Yuanzhen Bai1, Jia Tong2
1Nanjing University of Chinese Medicine, 210023, Nanjing, China.
Abstract:
Our paper aimed to disclose the effects of nobiletin (NOB) on hormone-induced osteoblast apoptosis and potential action mechanism. MC3T3-E1 cells were randomly separated into normal group, glucocorticoid (GC) group, L-NOB group, M-NOB group, H-NOB group, and Colivelin group (Colivelin: JAK2/STAT3 activator). CCK-8 was applied to ascertain the activity of MC3T3-E1 cells. FITC-Annexin V/PI method was applied to measure cell apoptosis. Alkaline phosphatase (ALP) assay kit was applied to measure ALP activity. Enzyme linked immunosorbent assay (ELISA) was implemented to ascertain the levels of IL-6, IL-1β, TNF-α, and ROS. Western blotting was implemented to distinguish the expressions of JAK2/STAT3 pathway proteins. The viability of MC3T3-E1 cells, ALP activities, and bcl-2 protein level were considerably decreased, while the apoptotic rate, the levels of TNF-α, IL-1β, IL-6, ROS, and the expressions of pJAK2/JAK2, pSTAT3/STAT3, Caspase-3, and bax proteins were greatly increased in each group after GC treatment. In comparison with GC group, MC3T3-E1 cell viability, ALP activity, and bcl-2 protein level in the L-NOB group, M-NOB group, and H-NOB group were greatly increased. Conversely, the apoptotic rate, the levels of TNF-α, IL-1β, IL-6, ROS, and the expressions of pJAK2/JAK2, pSTAT3/STAT3, Caspase-3, and bax proteins were markedly reduced. In contrast to H-NOB group, the apoptotic rate, the levels of TNF-α, IL-1β, IL-6, ROS, and the expressions of pJAK2/JAK2, pSTAT3/STAT3, Caspase-3, and bax proteins in Colivelin group were considerably enhanced, while MC3T3-E1 cell viability, ALP activity, and bcl-2 protein level were greatly declined. NOB ameliorates hormone-induced osteoblast apoptosis by reducing JAK2/STAT3 signaling activity.
Insights
Nobiletin (NOB) protects against hormone-induced osteoblast apoptosis by inhibiting the JAK2/STAT3 pathway. This natural compound enhances cell viability and reduces inflammatory markers, offering a potential therapeutic strategy for bone health.
Area of Science:
- Biochemistry
- Cell Biology
- Pharmacology
Background:
- Hormone-induced osteoblast apoptosis contributes to bone diseases.
- Glucocorticoids (GC) are known to induce osteoblast apoptosis.
- Nobiletin (NOB) is a flavonoid with potential therapeutic properties.
Purpose of the Study:
- To investigate the effects of nobiletin (NOB) on hormone-induced osteoblast apoptosis.
- To elucidate the underlying mechanism of NOB's action, particularly its effect on the JAK2/STAT3 pathway.
Main Methods:
- MC3T3-E1 osteoblast cell line was used.
- Cells were treated with glucocorticoid (GC) and varying concentrations of NOB.
- Cell viability (CCK-8), apoptosis (FITC-Annexin V/PI), ALP activity, inflammatory cytokine levels (IL-6, IL-1β, TNF-α), ROS levels, and JAK2/STAT3 pathway protein expression (Western blotting) were assessed.
Main Results:
- GC treatment increased osteoblast apoptosis, reduced cell viability and ALP activity, and elevated inflammatory markers (IL-6, IL-1β, TNF-α, ROS) and apoptosis-related proteins (Caspase-3, Bax).
- NOB treatment dose-dependently reversed these effects, increasing cell viability, ALP activity, and Bcl-2 levels, while decreasing apoptosis, inflammation, ROS, and downstream JAK2/STAT3 signaling.
- Activation of JAK2/STAT3 signaling with Colivelin counteracted NOB's protective effects.
Conclusions:
- Nobiletin (NOB) effectively ameliorates hormone-induced osteoblast apoptosis.
- NOB exerts its protective effects by inhibiting the JAK2/STAT3 signaling pathway.
- NOB demonstrates potential as a therapeutic agent for conditions involving osteoblast apoptosis.
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