Synergistic Inhibition of Pro-Inflammatory Pathways by Ginger and Turmeric Extracts in RAW 264.7 Cells
Synergy plays a prominent role in herbal medicines to increase potency and widen the therapeutic windows. The mechanism of synergy in herbal medicines is often associated with multi-targeted behavior and complex signaling pathways which are challenging to elucidate. This study aims to investigate th...
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| Format: | Article |
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Frontiers Media S.A.
2022-05-01
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| Series: | Frontiers in Pharmacology |
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| Online Access: | https://www.frontiersin.org/articles/10.3389/fphar.2022.818166/full |
| _version_ | 1818252933685313536 |
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| author | Xian Zhou Gerald Münch Hans Wohlmuth Hans Wohlmuth Hans Wohlmuth Sualiha Afzal Ming-Hui (Tim) Kao Ahmad Al-Khazaleh Mitchell Low David Leach Chun Guang Li |
| author_facet | Xian Zhou Gerald Münch Hans Wohlmuth Hans Wohlmuth Hans Wohlmuth Sualiha Afzal Ming-Hui (Tim) Kao Ahmad Al-Khazaleh Mitchell Low David Leach Chun Guang Li |
| author_sort | Xian Zhou |
| collection | DOAJ |
| description | Synergy plays a prominent role in herbal medicines to increase potency and widen the therapeutic windows. The mechanism of synergy in herbal medicines is often associated with multi-targeted behavior and complex signaling pathways which are challenging to elucidate. This study aims to investigate the synergistic mechanism of a combination (GT) of ginger (G) and turmeric (T) extracts by exploring the modulatory activity in lipopolysaccharides (LPS)-induced inflammatory pathways and key molecular targets. A Bioplex ProTM mouse cytokine 23-plex assay was utilized to assess the broad anti-cytokine activity of GT in LPS and interferon (IFN)-ɣ (both at 50 ng/mL)-activated RAW 264.7 cells. The inhibitory effects of individual and combined G and T on major proinflammatory mediators including nitric oxide (NO), tumor necrosis factor (TNF) and interleukin (IL)-6 were tested using Griess reagents and ELISA assays, respectively. Immunofluorescent staining and Western blot were used to investigate the modulatory effect of GT on key proteins in the LPS/TLR4 signaling transduction. The regulation of murine microRNA miR-155-5p was tested using real-time PCR. The IC50 value and combination index (CI) values were used to demonstrate potency and synergistic interaction, respectively. GT synergistically attenuated a range of pro-inflammatory mediators including inducible NO, major cytokines (TNF and IL-6) and secondary inflammatory cytokines (GM-CSF and MCP-1). GT significantly inhibited LPS-induced NF-kB p65 translocation, the activation of TLR4, TRAF6, and phosphorylation of JNK and c-JUN. Moreover, the suppressive effect of GT on each of the protein targets in this axis was stronger than that of the individual components. Real-time PCR analysis showed that GT suppressed miR-155-5p to a greater extent than G or T alone in LPS-stimulated cells. Our study demonstrates the synergistic mechanism of GT in downregulating LPS-induced proinflammatory pathways at the miRNA and protein levels. Our results establish a scientific basis for the combined application of G and T as an advanced therapeutic candidate in inflammatory diseases with broad and synergistic anti-inflammatory activity and multi-targeted mechanisms. |
| first_indexed | 2024-12-12T16:32:03Z |
| format | Article |
| id | doaj.art-60d686eda77a4a598853f6a30358c773 |
| institution | Directory Open Access Journal |
| issn | 1663-9812 |
| language | English |
| last_indexed | 2024-12-12T16:32:03Z |
| publishDate | 2022-05-01 |
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| spelling | doaj.art-60d686eda77a4a598853f6a30358c7732022-12-22T00:18:46ZengFrontiers Media S.A.Frontiers in Pharmacology1663-98122022-05-011310.3389/fphar.2022.818166818166Synergistic Inhibition of Pro-Inflammatory Pathways by Ginger and Turmeric Extracts in RAW 264.7 CellsXian Zhou0Gerald Münch1Hans Wohlmuth2Hans Wohlmuth3Hans Wohlmuth4Sualiha Afzal5Ming-Hui (Tim) Kao6Ahmad Al-Khazaleh7Mitchell Low8David Leach9Chun Guang Li10NICM Health Research Institute, Western Sydney University, Westmead, NSW, AustraliaSchool of Medicine, Western Sydney University, Campbelltown, NSW, AustraliaNICM Health Research Institute, Western Sydney University, Westmead, NSW, AustraliaIntegria Healthcare, Eight Mile Plains, QLD, AustraliaSchool of Chemistry and Molecular Biosciences, The University of Queensland, Brisbane, QLD, AustraliaSchool of Medicine, Western Sydney University, Campbelltown, NSW, AustraliaSchool of Medicine, Western Sydney University, Campbelltown, NSW, AustraliaNICM Health Research Institute, Western Sydney University, Westmead, NSW, AustraliaNICM Health Research Institute, Western Sydney University, Westmead, NSW, AustraliaIntegria Healthcare, Eight Mile Plains, QLD, AustraliaNICM Health Research Institute, Western Sydney University, Westmead, NSW, AustraliaSynergy plays a prominent role in herbal medicines to increase potency and widen the therapeutic windows. The mechanism of synergy in herbal medicines is often associated with multi-targeted behavior and complex signaling pathways which are challenging to elucidate. This study aims to investigate the synergistic mechanism of a combination (GT) of ginger (G) and turmeric (T) extracts by exploring the modulatory activity in lipopolysaccharides (LPS)-induced inflammatory pathways and key molecular targets. A Bioplex ProTM mouse cytokine 23-plex assay was utilized to assess the broad anti-cytokine activity of GT in LPS and interferon (IFN)-ɣ (both at 50 ng/mL)-activated RAW 264.7 cells. The inhibitory effects of individual and combined G and T on major proinflammatory mediators including nitric oxide (NO), tumor necrosis factor (TNF) and interleukin (IL)-6 were tested using Griess reagents and ELISA assays, respectively. Immunofluorescent staining and Western blot were used to investigate the modulatory effect of GT on key proteins in the LPS/TLR4 signaling transduction. The regulation of murine microRNA miR-155-5p was tested using real-time PCR. The IC50 value and combination index (CI) values were used to demonstrate potency and synergistic interaction, respectively. GT synergistically attenuated a range of pro-inflammatory mediators including inducible NO, major cytokines (TNF and IL-6) and secondary inflammatory cytokines (GM-CSF and MCP-1). GT significantly inhibited LPS-induced NF-kB p65 translocation, the activation of TLR4, TRAF6, and phosphorylation of JNK and c-JUN. Moreover, the suppressive effect of GT on each of the protein targets in this axis was stronger than that of the individual components. Real-time PCR analysis showed that GT suppressed miR-155-5p to a greater extent than G or T alone in LPS-stimulated cells. Our study demonstrates the synergistic mechanism of GT in downregulating LPS-induced proinflammatory pathways at the miRNA and protein levels. Our results establish a scientific basis for the combined application of G and T as an advanced therapeutic candidate in inflammatory diseases with broad and synergistic anti-inflammatory activity and multi-targeted mechanisms.https://www.frontiersin.org/articles/10.3389/fphar.2022.818166/fullgingerturmericsynergyMAPKTLR4-TRAF6-NFκBmiR-155-5p |
| spellingShingle | Xian Zhou Gerald Münch Hans Wohlmuth Hans Wohlmuth Hans Wohlmuth Sualiha Afzal Ming-Hui (Tim) Kao Ahmad Al-Khazaleh Mitchell Low David Leach Chun Guang Li Synergistic Inhibition of Pro-Inflammatory Pathways by Ginger and Turmeric Extracts in RAW 264.7 Cells Frontiers in Pharmacology ginger turmeric synergy MAPK TLR4-TRAF6-NFκB miR-155-5p |
| title | Synergistic Inhibition of Pro-Inflammatory Pathways by Ginger and Turmeric Extracts in RAW 264.7 Cells |
| title_full | Synergistic Inhibition of Pro-Inflammatory Pathways by Ginger and Turmeric Extracts in RAW 264.7 Cells |
| title_fullStr | Synergistic Inhibition of Pro-Inflammatory Pathways by Ginger and Turmeric Extracts in RAW 264.7 Cells |
| title_full_unstemmed | Synergistic Inhibition of Pro-Inflammatory Pathways by Ginger and Turmeric Extracts in RAW 264.7 Cells |
| title_short | Synergistic Inhibition of Pro-Inflammatory Pathways by Ginger and Turmeric Extracts in RAW 264.7 Cells |
| title_sort | synergistic inhibition of pro inflammatory pathways by ginger and turmeric extracts in raw 264 7 cells |
| topic | ginger turmeric synergy MAPK TLR4-TRAF6-NFκB miR-155-5p |
| url | https://www.frontiersin.org/articles/10.3389/fphar.2022.818166/full |
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