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Polymethoxyflavone Apigenin-Trimethylether Suppresses LPS-Induced Inflammatory Response in Nontransformed Porcine Intestinal Cell Line IPEC-J2.

Farkas O, Palócz O, Pászti-Gere E, Gálfi P - Oxid Med Cell Longev (2015)

Bottom Line: Treatment with both flavonoids caused significant reduction in the mRNA level of COX-2, but the anti-inflammatory effect of the methylated analogue was more effective than the unmethylated one.Furthermore, both flavonoids reduced significantly the level of extracellular H2O2 compared to the control cells.In conclusion, the methylated apigenin analogue could avoid LPS-induced intestinal inflammation and it could be applied in the future as an effective anti-inflammatory compound.

View Article: PubMed Central - PubMed

Affiliation: Department of Pharmacology and Toxicology, Faculty of Veterinary Science, Szent István University, István utca 2, Budapest 1078, Hungary.

ABSTRACT
The in vitro anti-inflammatory effect of apigenin and its trimethylated analogue (apigenin-trimethylether) has been investigated in order to evaluate whether these flavonoids could attenuate LPS-induced inflammation in IPEC-J2 non-transformed intestinal epithelial cells. Levels of IL-6, IL-8, TNF-α, and COX-2 mRNA were measured as a marker of inflammatory response. The extracellular H2O2 level in IPEC-J2 cells was also monitored by Amplex Red assay. Our data revealed that both compounds had significant lowering effect on the inflammatory response. Apigenin (at 25 μM) significantly decreased gene expression of IL-6 in LPS-treated cells, while apigenin-trimethylether in the same concentration did not influence IL-6 mRNA level. Both apigenin and apigenin-trimethylether reduced IL-8 gene expression significantly. TNF-α mRNA level was decreased by apigenin-trimethylether, which was not influenced by apigenin. Treatment with both flavonoids caused significant reduction in the mRNA level of COX-2, but the anti-inflammatory effect of the methylated analogue was more effective than the unmethylated one. Furthermore, both flavonoids reduced significantly the level of extracellular H2O2 compared to the control cells. In conclusion, the methylated apigenin analogue could avoid LPS-induced intestinal inflammation and it could be applied in the future as an effective anti-inflammatory compound.

No MeSH data available.


Related in: MedlinePlus

Level of extracellular H2O2 in IPEC-J2 cells exposed to flavonoid treatment (25 and 50 μM; 1 h). Effect of apigenin and apigenin-trimethylether on the relative extracellular H2O2 levels (n = 3-4/group; *P < 0.05). Fluorescence measurement was performed by Amplex Red method. Fluorescence was detected immediately (a) and 24 h after (b) flavonoid treatment. Data are shown as means + standard deviations. API = apigenin; API-TM = apigenin-trimethylether.
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fig7: Level of extracellular H2O2 in IPEC-J2 cells exposed to flavonoid treatment (25 and 50 μM; 1 h). Effect of apigenin and apigenin-trimethylether on the relative extracellular H2O2 levels (n = 3-4/group; *P < 0.05). Fluorescence measurement was performed by Amplex Red method. Fluorescence was detected immediately (a) and 24 h after (b) flavonoid treatment. Data are shown as means + standard deviations. API = apigenin; API-TM = apigenin-trimethylether.

Mentions: Figure 7 shows the relative extracellular H2O2 level in IPEC-J2 cells after flavonoid treatment. LPS only at higher concentration (at 50 μg/mL) increased the level of extracellular H2O2; however, IPEC-J2 cells are irreversibly damaged using LPS in this concentration (data not shown). Flavonoids showed different effects on the rate of H2O2 production, depending on the duration of incubation in DMEM after flavonoid treatment. In case of short time effect measurements (1 h incubation with flavonoids and detection immediately after the incubation period), neither apigenin nor its trimethylated analogue decreased mitochondrial H2O2 production rate effectively. Long time effects of flavonoid treatment (Amplex Red measurement was performed 24 h after the 1 h flavonoid incubation) were also studied. Extracellular H2O2 level was significantly decreased in case of both lower (25 μM) and higher (50 μM) concentration apigenin treatments. The same reducing effect was found, when 25 μM and 50 μM apigenin-trimethylether were applied.


Polymethoxyflavone Apigenin-Trimethylether Suppresses LPS-Induced Inflammatory Response in Nontransformed Porcine Intestinal Cell Line IPEC-J2.

Farkas O, Palócz O, Pászti-Gere E, Gálfi P - Oxid Med Cell Longev (2015)

Level of extracellular H2O2 in IPEC-J2 cells exposed to flavonoid treatment (25 and 50 μM; 1 h). Effect of apigenin and apigenin-trimethylether on the relative extracellular H2O2 levels (n = 3-4/group; *P < 0.05). Fluorescence measurement was performed by Amplex Red method. Fluorescence was detected immediately (a) and 24 h after (b) flavonoid treatment. Data are shown as means + standard deviations. API = apigenin; API-TM = apigenin-trimethylether.
© Copyright Policy - open-access
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC4477253&req=5

fig7: Level of extracellular H2O2 in IPEC-J2 cells exposed to flavonoid treatment (25 and 50 μM; 1 h). Effect of apigenin and apigenin-trimethylether on the relative extracellular H2O2 levels (n = 3-4/group; *P < 0.05). Fluorescence measurement was performed by Amplex Red method. Fluorescence was detected immediately (a) and 24 h after (b) flavonoid treatment. Data are shown as means + standard deviations. API = apigenin; API-TM = apigenin-trimethylether.
Mentions: Figure 7 shows the relative extracellular H2O2 level in IPEC-J2 cells after flavonoid treatment. LPS only at higher concentration (at 50 μg/mL) increased the level of extracellular H2O2; however, IPEC-J2 cells are irreversibly damaged using LPS in this concentration (data not shown). Flavonoids showed different effects on the rate of H2O2 production, depending on the duration of incubation in DMEM after flavonoid treatment. In case of short time effect measurements (1 h incubation with flavonoids and detection immediately after the incubation period), neither apigenin nor its trimethylated analogue decreased mitochondrial H2O2 production rate effectively. Long time effects of flavonoid treatment (Amplex Red measurement was performed 24 h after the 1 h flavonoid incubation) were also studied. Extracellular H2O2 level was significantly decreased in case of both lower (25 μM) and higher (50 μM) concentration apigenin treatments. The same reducing effect was found, when 25 μM and 50 μM apigenin-trimethylether were applied.

Bottom Line: Treatment with both flavonoids caused significant reduction in the mRNA level of COX-2, but the anti-inflammatory effect of the methylated analogue was more effective than the unmethylated one.Furthermore, both flavonoids reduced significantly the level of extracellular H2O2 compared to the control cells.In conclusion, the methylated apigenin analogue could avoid LPS-induced intestinal inflammation and it could be applied in the future as an effective anti-inflammatory compound.

View Article: PubMed Central - PubMed

Affiliation: Department of Pharmacology and Toxicology, Faculty of Veterinary Science, Szent István University, István utca 2, Budapest 1078, Hungary.

ABSTRACT
The in vitro anti-inflammatory effect of apigenin and its trimethylated analogue (apigenin-trimethylether) has been investigated in order to evaluate whether these flavonoids could attenuate LPS-induced inflammation in IPEC-J2 non-transformed intestinal epithelial cells. Levels of IL-6, IL-8, TNF-α, and COX-2 mRNA were measured as a marker of inflammatory response. The extracellular H2O2 level in IPEC-J2 cells was also monitored by Amplex Red assay. Our data revealed that both compounds had significant lowering effect on the inflammatory response. Apigenin (at 25 μM) significantly decreased gene expression of IL-6 in LPS-treated cells, while apigenin-trimethylether in the same concentration did not influence IL-6 mRNA level. Both apigenin and apigenin-trimethylether reduced IL-8 gene expression significantly. TNF-α mRNA level was decreased by apigenin-trimethylether, which was not influenced by apigenin. Treatment with both flavonoids caused significant reduction in the mRNA level of COX-2, but the anti-inflammatory effect of the methylated analogue was more effective than the unmethylated one. Furthermore, both flavonoids reduced significantly the level of extracellular H2O2 compared to the control cells. In conclusion, the methylated apigenin analogue could avoid LPS-induced intestinal inflammation and it could be applied in the future as an effective anti-inflammatory compound.

No MeSH data available.


Related in: MedlinePlus