Limits...
Sesquiterpene lactones inhibit advanced oxidation protein product-induced MCP-1 expression in podocytes via an IKK/NF-κB-dependent mechanism.

Zhao Y, Chen SJ, Wang JC, Niu HX, Jia QQ, Chen XW, Du XY, Lu L, Huang B, Zhang Q, Chen Y, Long HB - Oxid Med Cell Longev (2015)

Bottom Line: Sesquiterpene lactones (SLs), originally isolated from Tanacetum parthenium, have been reported to exhibit anti-inflammatory effects but few studies have examined their effects on DN.The IKK/NF-κB inhibitor parthenolide decreased AOPP-induced MCP-1 expression.Pretreatment with SLs inhibited MCP-1 mRNA and protein expression and suppressed IKKβ and NF-κB p65 phosphorylation and IκBα degradation.

View Article: PubMed Central - PubMed

Affiliation: Division of Nephrology, Zhujiang Hospital, Southern Medical University, Guangzhou, Guangdong 510280, China ; Department of Nephrology, Xiangyang Central Hospital, Hubei University of Arts and Science, Xiangyang, Hubei 441021, China.

ABSTRACT
Inflammation is a relevant factor in the pathogenesis of diabetes nephropathy (DN). Sesquiterpene lactones (SLs), originally isolated from Tanacetum parthenium, have been reported to exhibit anti-inflammatory effects but few studies have examined their effects on DN. To determine whether advanced oxidation protein products (AOPPs) can induce the expression of chemokine monocyte chemoattractant protein- (MCP-) 1 in cultured mouse podocytes and to explore the mechanisms of the potential renoprotection of SLs, we treated podocytes with AOPPs and SLs (parthenolide and its derivatives micheliolide, compound 1, and compound 2). MCP-1 mRNA and protein expression were tested using quantitative real-time PCR and ELISA, respectively, and the protein levels of IKKβ, phospho-IKKβ, IκBα, NF-κB p65, phospho-NF-κB p65, and tubulin were analyzed by Western blotting. AOPPs activated the expression of MCP-1 mRNA and protein in a dose- and time-dependent manner, activated IKKβ and NF-κB p65, and promoted IκBα degradation. The IKK/NF-κB inhibitor parthenolide decreased AOPP-induced MCP-1 expression. Pretreatment with SLs inhibited MCP-1 mRNA and protein expression and suppressed IKKβ and NF-κB p65 phosphorylation and IκBα degradation. Taken together, these findings provide a novel explanation for the anti-inflammatory effects of SLs that will ultimately benefit DN and potentially other inflammatory and immune renal diseases.

Show MeSH

Related in: MedlinePlus

Compound 1 prevented AOPP-induced IKKβ and NF-κB p65 activation and IκBα degradation in a dose- and time-dependent manner in podocytes. After 24 h of serum starvation, podocytes were pretreated with compound 1 (5 μM, 10 μM, and 20 μM) for 1 h and then exposed to 200 μg/mL AOPPs or native MSA for 30 min. PTL (10 μM) was used as a positive control (a, b, and c). Alternatively, podocytes were treated with 200 μg/mL AOPPs for 30 min, and then compound 1 (20 μM) was added for a preset time (d, e, f). The protein expression of p-IKKα/β, p-NF-κB p65, and IκBα was analyzed by Western blotting. The data are expressed as the means ± SEM of three independent experiments. ANOVA, ##P < 0.01, ###P < 0.001 versus CON; *P < 0.05, **P < 0.01 versus AOPP. CON, untreated cells; MSA, mouse serum albumin.
© Copyright Policy - open-access
Related In: Results  -  Collection


getmorefigures.php?uid=PMC4309307&req=5

fig7: Compound 1 prevented AOPP-induced IKKβ and NF-κB p65 activation and IκBα degradation in a dose- and time-dependent manner in podocytes. After 24 h of serum starvation, podocytes were pretreated with compound 1 (5 μM, 10 μM, and 20 μM) for 1 h and then exposed to 200 μg/mL AOPPs or native MSA for 30 min. PTL (10 μM) was used as a positive control (a, b, and c). Alternatively, podocytes were treated with 200 μg/mL AOPPs for 30 min, and then compound 1 (20 μM) was added for a preset time (d, e, f). The protein expression of p-IKKα/β, p-NF-κB p65, and IκBα was analyzed by Western blotting. The data are expressed as the means ± SEM of three independent experiments. ANOVA, ##P < 0.01, ###P < 0.001 versus CON; *P < 0.05, **P < 0.01 versus AOPP. CON, untreated cells; MSA, mouse serum albumin.

Mentions: To further analyze whether factors of the NF-κB activation cascade are influenced by SLs, we pretreated podocytes with different concentrations of MCL, compound 1, and compound 2 for 1 h before incubation with 200 μg/mL of AOPPs or native MSA for 30 min or 200 μg/mL AOPPs for 30 min, and we also added SLs for the indicated times. As shown in Figures 6(a), 6(b), and 6(c), Figures 7(a), 7(b), and 7(c), and Figures 8(a), 8(b), and 8(c), MCL, compound 1, and compound 2 prevented the protein expression of phospho-IKKα/β and phospho-NF-κB p65 and increased the protein expression of IκBα in a dose-dependent manner. The effect of MCL, compound 1, and compound 2 on inhibiting the activation of IKKβ, degradation of IκBα, and activation of NF-κB peaked at 10 μM, 20 μM, and 5 μM, respectively, and almost reached the effect of 10 μM of PTL. Figures 6(d), 6(e), and 6(f), Figures 7(d), 7(e), and 7(f), and Figures 8(d), 8(e), and 8(f) suggest that these three compounds prevented the phosphorylation of IKKβ and NF-κB p65 and the degradation of IκBα protein in a time-dependent manner.


Sesquiterpene lactones inhibit advanced oxidation protein product-induced MCP-1 expression in podocytes via an IKK/NF-κB-dependent mechanism.

Zhao Y, Chen SJ, Wang JC, Niu HX, Jia QQ, Chen XW, Du XY, Lu L, Huang B, Zhang Q, Chen Y, Long HB - Oxid Med Cell Longev (2015)

Compound 1 prevented AOPP-induced IKKβ and NF-κB p65 activation and IκBα degradation in a dose- and time-dependent manner in podocytes. After 24 h of serum starvation, podocytes were pretreated with compound 1 (5 μM, 10 μM, and 20 μM) for 1 h and then exposed to 200 μg/mL AOPPs or native MSA for 30 min. PTL (10 μM) was used as a positive control (a, b, and c). Alternatively, podocytes were treated with 200 μg/mL AOPPs for 30 min, and then compound 1 (20 μM) was added for a preset time (d, e, f). The protein expression of p-IKKα/β, p-NF-κB p65, and IκBα was analyzed by Western blotting. The data are expressed as the means ± SEM of three independent experiments. ANOVA, ##P < 0.01, ###P < 0.001 versus CON; *P < 0.05, **P < 0.01 versus AOPP. CON, untreated cells; MSA, mouse serum albumin.
© Copyright Policy - open-access
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC4309307&req=5

fig7: Compound 1 prevented AOPP-induced IKKβ and NF-κB p65 activation and IκBα degradation in a dose- and time-dependent manner in podocytes. After 24 h of serum starvation, podocytes were pretreated with compound 1 (5 μM, 10 μM, and 20 μM) for 1 h and then exposed to 200 μg/mL AOPPs or native MSA for 30 min. PTL (10 μM) was used as a positive control (a, b, and c). Alternatively, podocytes were treated with 200 μg/mL AOPPs for 30 min, and then compound 1 (20 μM) was added for a preset time (d, e, f). The protein expression of p-IKKα/β, p-NF-κB p65, and IκBα was analyzed by Western blotting. The data are expressed as the means ± SEM of three independent experiments. ANOVA, ##P < 0.01, ###P < 0.001 versus CON; *P < 0.05, **P < 0.01 versus AOPP. CON, untreated cells; MSA, mouse serum albumin.
Mentions: To further analyze whether factors of the NF-κB activation cascade are influenced by SLs, we pretreated podocytes with different concentrations of MCL, compound 1, and compound 2 for 1 h before incubation with 200 μg/mL of AOPPs or native MSA for 30 min or 200 μg/mL AOPPs for 30 min, and we also added SLs for the indicated times. As shown in Figures 6(a), 6(b), and 6(c), Figures 7(a), 7(b), and 7(c), and Figures 8(a), 8(b), and 8(c), MCL, compound 1, and compound 2 prevented the protein expression of phospho-IKKα/β and phospho-NF-κB p65 and increased the protein expression of IκBα in a dose-dependent manner. The effect of MCL, compound 1, and compound 2 on inhibiting the activation of IKKβ, degradation of IκBα, and activation of NF-κB peaked at 10 μM, 20 μM, and 5 μM, respectively, and almost reached the effect of 10 μM of PTL. Figures 6(d), 6(e), and 6(f), Figures 7(d), 7(e), and 7(f), and Figures 8(d), 8(e), and 8(f) suggest that these three compounds prevented the phosphorylation of IKKβ and NF-κB p65 and the degradation of IκBα protein in a time-dependent manner.

Bottom Line: Sesquiterpene lactones (SLs), originally isolated from Tanacetum parthenium, have been reported to exhibit anti-inflammatory effects but few studies have examined their effects on DN.The IKK/NF-κB inhibitor parthenolide decreased AOPP-induced MCP-1 expression.Pretreatment with SLs inhibited MCP-1 mRNA and protein expression and suppressed IKKβ and NF-κB p65 phosphorylation and IκBα degradation.

View Article: PubMed Central - PubMed

Affiliation: Division of Nephrology, Zhujiang Hospital, Southern Medical University, Guangzhou, Guangdong 510280, China ; Department of Nephrology, Xiangyang Central Hospital, Hubei University of Arts and Science, Xiangyang, Hubei 441021, China.

ABSTRACT
Inflammation is a relevant factor in the pathogenesis of diabetes nephropathy (DN). Sesquiterpene lactones (SLs), originally isolated from Tanacetum parthenium, have been reported to exhibit anti-inflammatory effects but few studies have examined their effects on DN. To determine whether advanced oxidation protein products (AOPPs) can induce the expression of chemokine monocyte chemoattractant protein- (MCP-) 1 in cultured mouse podocytes and to explore the mechanisms of the potential renoprotection of SLs, we treated podocytes with AOPPs and SLs (parthenolide and its derivatives micheliolide, compound 1, and compound 2). MCP-1 mRNA and protein expression were tested using quantitative real-time PCR and ELISA, respectively, and the protein levels of IKKβ, phospho-IKKβ, IκBα, NF-κB p65, phospho-NF-κB p65, and tubulin were analyzed by Western blotting. AOPPs activated the expression of MCP-1 mRNA and protein in a dose- and time-dependent manner, activated IKKβ and NF-κB p65, and promoted IκBα degradation. The IKK/NF-κB inhibitor parthenolide decreased AOPP-induced MCP-1 expression. Pretreatment with SLs inhibited MCP-1 mRNA and protein expression and suppressed IKKβ and NF-κB p65 phosphorylation and IκBα degradation. Taken together, these findings provide a novel explanation for the anti-inflammatory effects of SLs that will ultimately benefit DN and potentially other inflammatory and immune renal diseases.

Show MeSH
Related in: MedlinePlus