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Green tea polyphenols attenuate LPS-induced inflammation through upregulating microRNA-9 in murine chondrogenic ATDC5
Qiao Zhang1, Yongkun Wang1, Mingran Zhang1
1Department of Orthopaedics, China-Japan Union Hospital of Jilin University, Changchun, Jilin, China.
Background:
Osteoarthritis (OA), a universal chronic musculoskeletal disorder, is closely related to inflammation. More effective drugs for improving OA outcome are definitely needed. Herein, we attempted to verify the protective role of green tea polyphenols (GTP) after treatment with murine in ATDC5 cells to reveal the regulatory mechanism.
Methods:
ATDC5 cells were stimulated with lipopolysaccharide (LPS) to mimic an inflammatory response during OA. Cell activity, apoptosis, levels of relative proteins, and prophlogistic factors were tested via a Cell Counting Kit-8 experiment, a flow cytometry experiment, western blot, and RT-qPCR (ELISA and Western blot), separately. miR-9 level was detected by RT-qPCR and altered via miR-9 mimic and inhibitor transfection. We finally studied MAPK and NF-κB pathways in GTP-related modulations using western blot.
Results:
LPS caused inflammatory cell damage in ATDC5 cells, showing decreased cell activity, enhanced apoptosis, and increased levels of pro-inflammatory cytokines. GTP pretreatments could significantly attenuate LPS-induced alterations. In addition, LPS-induced miR-9 upregulation was further positively regulated in ATDC5 cells. The effects of GTP pretreatments in LPS-caused ATDC5 cells were enhanced via miR-9 upregulation, whereas they were reduced via miR-9 suppression. Finally, we found that GTP pretreatments could suppress the MAPK and NF-κB pathways through miR-9 regulation.
Conclusion:
GTP pretreatments attenuated LPS-induced inflammatory response accompanied by the suppression of the MAPK and NF-κB pathways via positively regulating miR-9 in ATDC5 cells.
Insights
Green tea polyphenols (GTP) protect against osteoarthritis inflammation by upregulating miR-9, which suppresses key inflammatory pathways like MAPK and NF-κB in ATDC5 cells.
Area of Science:
- Biochemistry
- Cell Biology
- Pharmacology
Background:
- Osteoarthritis (OA) is a widespread chronic musculoskeletal disorder linked to inflammation.
- There is a critical need for improved therapeutic agents to manage OA outcomes.
- This study investigates the protective effects of green tea polyphenols (GTP) in an OA model.
Purpose of the Study:
- To verify the protective role of green tea polyphenols (GTP) in an in vitro osteoarthritis model.
- To elucidate the regulatory mechanisms underlying GTP's effects on inflammation.
- To explore the involvement of miR-9 and associated signaling pathways.
Main Methods:
- ATDC5 cells were stimulated with lipopolysaccharide (LPS) to induce an inflammatory response.
- Cell viability, apoptosis, protein levels, and pro-inflammatory factors were assessed.
- miR-9 levels were modulated using mimics and inhibitors, and signaling pathways (MAPK, NF-κB) were analyzed.
Main Results:
- LPS induced inflammatory damage in ATDC5 cells, characterized by reduced viability and increased apoptosis and cytokine levels.
- GTP pretreatment significantly mitigated LPS-induced inflammatory damage.
- GTP enhanced miR-9 levels, which in turn suppressed MAPK and NF-κB pathways, mediating the protective effects.
Conclusions:
- Green tea polyphenols (GTP) demonstrate a protective effect against LPS-induced inflammation in ATDC5 cells.
- GTP exerts its anti-inflammatory action by positively regulating miR-9.
- This miR-9 regulation leads to the suppression of the MAPK and NF-κB signaling pathways, offering a potential therapeutic strategy for OA.
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