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Anti-inflammatory effects of cordycepin in lipopolysaccharide-stimulated RAW 264.7 macrophages through Toll-like receptor 4-mediated suppression of mitogen-activated protein kinases and NF-κB signaling pathways.

Choi YH, Kim GY, Lee HH - Drug Des Devel Ther (2014)

Bottom Line: This compound possesses many pharmacological properties, such as an ability to enhance immune function, as well as antioxidant, antiaging, and anticancer effects.In addition, pretreatment with cordycepin significantly inhibited lipopolysaccharide (LPS)-induced phosphorylation of mitogen-activating protein kinases and attenuated nuclear translocation of NF-κB by LPS, which was associated with abrogation of inhibitor kappa B-alpha degradation.Taken together, the results suggest that the inhibitory effects of cordycepin on LPS-stimulated inflammatory responses in RAW 264.7 macrophages are associated with suppression of mitogen-activating protein kinases and activation of NF-κB by inhibition of the Toll-like receptor 4 signaling pathway.

View Article: PubMed Central - PubMed

Affiliation: Department of Biochemistry, Dongeui University College of Korean Medicine, Busan, Republic of Korea ; Anti-Aging Research Center and Blue-Bio Industry RIC, Dongeui University, Busan, Republic of Korea.

ABSTRACT
Cordycepin is the main functional component of the Cordyceps species, which has been widely used in traditional Oriental medicine. This compound possesses many pharmacological properties, such as an ability to enhance immune function, as well as antioxidant, antiaging, and anticancer effects. In the present study, we investigated the anti-inflammatory effects of cordycepin using a murine macrophage RAW 264.7 cell model. Our data demonstrated that cordycepin suppressed production of proinflammatory mediators such as nitric oxide (NO) and prostaglandin E2 by inhibiting inducible NO synthase and cyclooxygenase-2 gene expression. Cordycepin also inhibited the release of proinflammatory cytokines, including tumor necrosis factor-alpha and interleukin-1-beta, through downregulation of respective mRNA expression. In addition, pretreatment with cordycepin significantly inhibited lipopolysaccharide (LPS)-induced phosphorylation of mitogen-activating protein kinases and attenuated nuclear translocation of NF-κB by LPS, which was associated with abrogation of inhibitor kappa B-alpha degradation. Furthermore, cordycepin potently inhibited the binding of LPS to macrophages and LPS-induced Toll-like receptor 4 and myeloid differentiation factor 88 expression. Taken together, the results suggest that the inhibitory effects of cordycepin on LPS-stimulated inflammatory responses in RAW 264.7 macrophages are associated with suppression of mitogen-activating protein kinases and activation of NF-κB by inhibition of the Toll-like receptor 4 signaling pathway.

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Effects of the TLR4 inhibitor CLI-095 on nitric oxide and PGE2 production in LPS-stimulated RAW 264.7 macrophages.Notes: (A, B) Cells were treated with 30 μg/mL cordycepin alone or in combination with 15 μM CLI-095 for 1 hour before LPS treatment. Following 24 hours of treatment, the amounts of nitric oxide and PGE2 production were measured with the supernatants. The data are shown as the mean ± standard deviation of three independent experiments (*P<0.05 versus LPS treated cells; #P<0.05 versus cells treated with LPS plus cordycepin). The levels of iNOS and COX-2 mRNA (C) and protein (D) were assessed by reverse transcriptase polymerase chain reaction and Western blot assays after 6 hours and 24 hours of treatment, respectively. GAPDH and actin were used as internal controls, respectively.Abbreviations: GAPDH, glyceraldehyde-3-phosphate dehydrogenase; LPS, lipopolysaccharide; iNOS, inducible nitric oxide synthase; PGE2, prostaglandin E2;COX-2, cyclooxygenase-2.
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f6-dddt-8-1941: Effects of the TLR4 inhibitor CLI-095 on nitric oxide and PGE2 production in LPS-stimulated RAW 264.7 macrophages.Notes: (A, B) Cells were treated with 30 μg/mL cordycepin alone or in combination with 15 μM CLI-095 for 1 hour before LPS treatment. Following 24 hours of treatment, the amounts of nitric oxide and PGE2 production were measured with the supernatants. The data are shown as the mean ± standard deviation of three independent experiments (*P<0.05 versus LPS treated cells; #P<0.05 versus cells treated with LPS plus cordycepin). The levels of iNOS and COX-2 mRNA (C) and protein (D) were assessed by reverse transcriptase polymerase chain reaction and Western blot assays after 6 hours and 24 hours of treatment, respectively. GAPDH and actin were used as internal controls, respectively.Abbreviations: GAPDH, glyceraldehyde-3-phosphate dehydrogenase; LPS, lipopolysaccharide; iNOS, inducible nitric oxide synthase; PGE2, prostaglandin E2;COX-2, cyclooxygenase-2.

Mentions: To confirm involvement of the TLR4 signaling pathway in the cordycepin-mediated anti-inflammatory potential, we next evaluated the effects of the pharmacological agent CLI-095, which blocks TLR4-mediated signaling. As shown in Figure 6, we found that cordycepin and CLI-095 cotreatment synergistically inhibited LPS-induced production of NO and PGE2 as well as iNOS and COX-2 expression. Furthermore, the effects of LPS-enhanced TNF-α and IL-1β release were successfully inhibited by blocking TLR4 signaling with CLI-095, and cotreatment of cordycepin with CLI-095 almost completely inhibited the production of those proinflammatory cytokines to background levels similar to that in the untreated control (Figure 7). Taken together, these results verify that the inhibitory effects of cordycepin on the LPS-induced inflammatory response results, at least, from suppression of TLR4 signaling cascades in RAW 264.7 macrophages.


Anti-inflammatory effects of cordycepin in lipopolysaccharide-stimulated RAW 264.7 macrophages through Toll-like receptor 4-mediated suppression of mitogen-activated protein kinases and NF-κB signaling pathways.

Choi YH, Kim GY, Lee HH - Drug Des Devel Ther (2014)

Effects of the TLR4 inhibitor CLI-095 on nitric oxide and PGE2 production in LPS-stimulated RAW 264.7 macrophages.Notes: (A, B) Cells were treated with 30 μg/mL cordycepin alone or in combination with 15 μM CLI-095 for 1 hour before LPS treatment. Following 24 hours of treatment, the amounts of nitric oxide and PGE2 production were measured with the supernatants. The data are shown as the mean ± standard deviation of three independent experiments (*P<0.05 versus LPS treated cells; #P<0.05 versus cells treated with LPS plus cordycepin). The levels of iNOS and COX-2 mRNA (C) and protein (D) were assessed by reverse transcriptase polymerase chain reaction and Western blot assays after 6 hours and 24 hours of treatment, respectively. GAPDH and actin were used as internal controls, respectively.Abbreviations: GAPDH, glyceraldehyde-3-phosphate dehydrogenase; LPS, lipopolysaccharide; iNOS, inducible nitric oxide synthase; PGE2, prostaglandin E2;COX-2, cyclooxygenase-2.
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4206205&req=5

f6-dddt-8-1941: Effects of the TLR4 inhibitor CLI-095 on nitric oxide and PGE2 production in LPS-stimulated RAW 264.7 macrophages.Notes: (A, B) Cells were treated with 30 μg/mL cordycepin alone or in combination with 15 μM CLI-095 for 1 hour before LPS treatment. Following 24 hours of treatment, the amounts of nitric oxide and PGE2 production were measured with the supernatants. The data are shown as the mean ± standard deviation of three independent experiments (*P<0.05 versus LPS treated cells; #P<0.05 versus cells treated with LPS plus cordycepin). The levels of iNOS and COX-2 mRNA (C) and protein (D) were assessed by reverse transcriptase polymerase chain reaction and Western blot assays after 6 hours and 24 hours of treatment, respectively. GAPDH and actin were used as internal controls, respectively.Abbreviations: GAPDH, glyceraldehyde-3-phosphate dehydrogenase; LPS, lipopolysaccharide; iNOS, inducible nitric oxide synthase; PGE2, prostaglandin E2;COX-2, cyclooxygenase-2.
Mentions: To confirm involvement of the TLR4 signaling pathway in the cordycepin-mediated anti-inflammatory potential, we next evaluated the effects of the pharmacological agent CLI-095, which blocks TLR4-mediated signaling. As shown in Figure 6, we found that cordycepin and CLI-095 cotreatment synergistically inhibited LPS-induced production of NO and PGE2 as well as iNOS and COX-2 expression. Furthermore, the effects of LPS-enhanced TNF-α and IL-1β release were successfully inhibited by blocking TLR4 signaling with CLI-095, and cotreatment of cordycepin with CLI-095 almost completely inhibited the production of those proinflammatory cytokines to background levels similar to that in the untreated control (Figure 7). Taken together, these results verify that the inhibitory effects of cordycepin on the LPS-induced inflammatory response results, at least, from suppression of TLR4 signaling cascades in RAW 264.7 macrophages.

Bottom Line: This compound possesses many pharmacological properties, such as an ability to enhance immune function, as well as antioxidant, antiaging, and anticancer effects.In addition, pretreatment with cordycepin significantly inhibited lipopolysaccharide (LPS)-induced phosphorylation of mitogen-activating protein kinases and attenuated nuclear translocation of NF-κB by LPS, which was associated with abrogation of inhibitor kappa B-alpha degradation.Taken together, the results suggest that the inhibitory effects of cordycepin on LPS-stimulated inflammatory responses in RAW 264.7 macrophages are associated with suppression of mitogen-activating protein kinases and activation of NF-κB by inhibition of the Toll-like receptor 4 signaling pathway.

View Article: PubMed Central - PubMed

Affiliation: Department of Biochemistry, Dongeui University College of Korean Medicine, Busan, Republic of Korea ; Anti-Aging Research Center and Blue-Bio Industry RIC, Dongeui University, Busan, Republic of Korea.

ABSTRACT
Cordycepin is the main functional component of the Cordyceps species, which has been widely used in traditional Oriental medicine. This compound possesses many pharmacological properties, such as an ability to enhance immune function, as well as antioxidant, antiaging, and anticancer effects. In the present study, we investigated the anti-inflammatory effects of cordycepin using a murine macrophage RAW 264.7 cell model. Our data demonstrated that cordycepin suppressed production of proinflammatory mediators such as nitric oxide (NO) and prostaglandin E2 by inhibiting inducible NO synthase and cyclooxygenase-2 gene expression. Cordycepin also inhibited the release of proinflammatory cytokines, including tumor necrosis factor-alpha and interleukin-1-beta, through downregulation of respective mRNA expression. In addition, pretreatment with cordycepin significantly inhibited lipopolysaccharide (LPS)-induced phosphorylation of mitogen-activating protein kinases and attenuated nuclear translocation of NF-κB by LPS, which was associated with abrogation of inhibitor kappa B-alpha degradation. Furthermore, cordycepin potently inhibited the binding of LPS to macrophages and LPS-induced Toll-like receptor 4 and myeloid differentiation factor 88 expression. Taken together, the results suggest that the inhibitory effects of cordycepin on LPS-stimulated inflammatory responses in RAW 264.7 macrophages are associated with suppression of mitogen-activating protein kinases and activation of NF-κB by inhibition of the Toll-like receptor 4 signaling pathway.

Show MeSH
Related in: MedlinePlus