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Effects of Chinese Propolis in Protecting Bovine Mammary Epithelial Cells against Mastitis Pathogens-Induced Cell Damage.

Wang K, Jin XL, Shen XG, Sun LP, Wu LM, Wei JQ, Marcucci MC, Hu FL, Liu JX - Mediators Inflamm. (2016)

Bottom Line: MAC-T cells treated with bacterial endotoxin (lipopolysaccharide, LPS), heat-inactivated Escherichia coli, and Staphylococcus aureus exhibited significant decreases in cell viability while TNF-α and lipoteichoic acid (LTA) did not.There were also corresponding decreases in expressions of proinflammatory IL-6 and TNF-α mRNA.CP and its polyphenolic active components (primarily caffeic acid phenethyl ester and quercetin) had strong inhibitive effects against NF-κB activation and increased the transcriptional activity of Nrf2-ARE.

View Article: PubMed Central - PubMed

Affiliation: College of Animal Sciences, Zhejiang University, Hangzhou 310058, China.

ABSTRACT
Chinese propolis (CP), an important hive product, can alleviate inflammatory responses. However, little is known regarding the potential of propolis treatment for mastitis control. To investigate the anti-inflammatory effects of CP on bovine mammary epithelial cells (MAC-T), we used a range of pathogens to induce cellular inflammatory damage. Cell viability was determined and expressions of inflammatory/antioxidant genes were measured. Using a cell-based reporter assay system, we evaluated CP and its primary constituents on the NF-κB and Nrf2-ARE transcription activation. MAC-T cells treated with bacterial endotoxin (lipopolysaccharide, LPS), heat-inactivated Escherichia coli, and Staphylococcus aureus exhibited significant decreases in cell viability while TNF-α and lipoteichoic acid (LTA) did not. Pretreatment with CP prevented losses in cell viability associated with the addition of killed bacteria or bacterial endotoxins. There were also corresponding decreases in expressions of proinflammatory IL-6 and TNF-α mRNA. Compared with the mastitis challenged cells, enhanced expressions of antioxidant genes HO-1, Txnrd-1, and GCLM were observed in CP-treated cells. CP and its polyphenolic active components (primarily caffeic acid phenethyl ester and quercetin) had strong inhibitive effects against NF-κB activation and increased the transcriptional activity of Nrf2-ARE. These findings suggest that propolis may be valuable in the control of bovine mastitis.

No MeSH data available.


Related in: MedlinePlus

Time and dose effects of CP treatment on inflammatory cytokine and cellular antioxidant defense gene expressions in TNF-α stimulated MAC-T cells. MAC-T cells were preincubated with different concentrations of propolis for 1 h and then stimulated with 25 ng/mL TNF-α for various time periods (time effects, right) or preincubated with various concentrations of propolis (dose effect, left) and then stimulated with 25 ng/mL TNF-α for 6 h. IL-6 (a), IL-8 (b), TNF-α (c), HO-1 (d), Txnrd-1 (e), and GCLM (f) mRNA expressions were quantified by real-time PCR. Ct values of target genes were normalized to the value of β-actin. Relative gene expression in TNF-α stimulation for 6 h was arbitrarily set to one. The data are shown as mean ± SD from three independent experiments. #P < 0.05 compared to the normal control and ∗significantly different from the TNF-α treated control (P < 0.05). ∗P < 0.05, ∗∗P < 0.01, and ∗∗∗P < 0.001.
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fig4: Time and dose effects of CP treatment on inflammatory cytokine and cellular antioxidant defense gene expressions in TNF-α stimulated MAC-T cells. MAC-T cells were preincubated with different concentrations of propolis for 1 h and then stimulated with 25 ng/mL TNF-α for various time periods (time effects, right) or preincubated with various concentrations of propolis (dose effect, left) and then stimulated with 25 ng/mL TNF-α for 6 h. IL-6 (a), IL-8 (b), TNF-α (c), HO-1 (d), Txnrd-1 (e), and GCLM (f) mRNA expressions were quantified by real-time PCR. Ct values of target genes were normalized to the value of β-actin. Relative gene expression in TNF-α stimulation for 6 h was arbitrarily set to one. The data are shown as mean ± SD from three independent experiments. #P < 0.05 compared to the normal control and ∗significantly different from the TNF-α treated control (P < 0.05). ∗P < 0.05, ∗∗P < 0.01, and ∗∗∗P < 0.001.

Mentions: As shown in Figure 4, the increased expressions of proinflammatory cytokines IL-6 (Figure 4(a)) and TNF-α (Figure 4(c)) were suppressed by CP in a time- and dose-dependent manner. As for another important chemokine, IL-8, its expression was significantly driven by TNF-α and immediately peaked at 3 h after TNF stimulus (Figure 4(b)). CP treatment strongly promoted IL-8 expression, which reached its peak after 9 h TNF stimulus. We also found that TNF-α had very limited effects on antioxidant gene expressions, HO-1, TXRND, and GCLM (Figures 4(d)–4(f)).


Effects of Chinese Propolis in Protecting Bovine Mammary Epithelial Cells against Mastitis Pathogens-Induced Cell Damage.

Wang K, Jin XL, Shen XG, Sun LP, Wu LM, Wei JQ, Marcucci MC, Hu FL, Liu JX - Mediators Inflamm. (2016)

Time and dose effects of CP treatment on inflammatory cytokine and cellular antioxidant defense gene expressions in TNF-α stimulated MAC-T cells. MAC-T cells were preincubated with different concentrations of propolis for 1 h and then stimulated with 25 ng/mL TNF-α for various time periods (time effects, right) or preincubated with various concentrations of propolis (dose effect, left) and then stimulated with 25 ng/mL TNF-α for 6 h. IL-6 (a), IL-8 (b), TNF-α (c), HO-1 (d), Txnrd-1 (e), and GCLM (f) mRNA expressions were quantified by real-time PCR. Ct values of target genes were normalized to the value of β-actin. Relative gene expression in TNF-α stimulation for 6 h was arbitrarily set to one. The data are shown as mean ± SD from three independent experiments. #P < 0.05 compared to the normal control and ∗significantly different from the TNF-α treated control (P < 0.05). ∗P < 0.05, ∗∗P < 0.01, and ∗∗∗P < 0.001.
© Copyright Policy - open-access
Related In: Results  -  Collection

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fig4: Time and dose effects of CP treatment on inflammatory cytokine and cellular antioxidant defense gene expressions in TNF-α stimulated MAC-T cells. MAC-T cells were preincubated with different concentrations of propolis for 1 h and then stimulated with 25 ng/mL TNF-α for various time periods (time effects, right) or preincubated with various concentrations of propolis (dose effect, left) and then stimulated with 25 ng/mL TNF-α for 6 h. IL-6 (a), IL-8 (b), TNF-α (c), HO-1 (d), Txnrd-1 (e), and GCLM (f) mRNA expressions were quantified by real-time PCR. Ct values of target genes were normalized to the value of β-actin. Relative gene expression in TNF-α stimulation for 6 h was arbitrarily set to one. The data are shown as mean ± SD from three independent experiments. #P < 0.05 compared to the normal control and ∗significantly different from the TNF-α treated control (P < 0.05). ∗P < 0.05, ∗∗P < 0.01, and ∗∗∗P < 0.001.
Mentions: As shown in Figure 4, the increased expressions of proinflammatory cytokines IL-6 (Figure 4(a)) and TNF-α (Figure 4(c)) were suppressed by CP in a time- and dose-dependent manner. As for another important chemokine, IL-8, its expression was significantly driven by TNF-α and immediately peaked at 3 h after TNF stimulus (Figure 4(b)). CP treatment strongly promoted IL-8 expression, which reached its peak after 9 h TNF stimulus. We also found that TNF-α had very limited effects on antioxidant gene expressions, HO-1, TXRND, and GCLM (Figures 4(d)–4(f)).

Bottom Line: MAC-T cells treated with bacterial endotoxin (lipopolysaccharide, LPS), heat-inactivated Escherichia coli, and Staphylococcus aureus exhibited significant decreases in cell viability while TNF-α and lipoteichoic acid (LTA) did not.There were also corresponding decreases in expressions of proinflammatory IL-6 and TNF-α mRNA.CP and its polyphenolic active components (primarily caffeic acid phenethyl ester and quercetin) had strong inhibitive effects against NF-κB activation and increased the transcriptional activity of Nrf2-ARE.

View Article: PubMed Central - PubMed

Affiliation: College of Animal Sciences, Zhejiang University, Hangzhou 310058, China.

ABSTRACT
Chinese propolis (CP), an important hive product, can alleviate inflammatory responses. However, little is known regarding the potential of propolis treatment for mastitis control. To investigate the anti-inflammatory effects of CP on bovine mammary epithelial cells (MAC-T), we used a range of pathogens to induce cellular inflammatory damage. Cell viability was determined and expressions of inflammatory/antioxidant genes were measured. Using a cell-based reporter assay system, we evaluated CP and its primary constituents on the NF-κB and Nrf2-ARE transcription activation. MAC-T cells treated with bacterial endotoxin (lipopolysaccharide, LPS), heat-inactivated Escherichia coli, and Staphylococcus aureus exhibited significant decreases in cell viability while TNF-α and lipoteichoic acid (LTA) did not. Pretreatment with CP prevented losses in cell viability associated with the addition of killed bacteria or bacterial endotoxins. There were also corresponding decreases in expressions of proinflammatory IL-6 and TNF-α mRNA. Compared with the mastitis challenged cells, enhanced expressions of antioxidant genes HO-1, Txnrd-1, and GCLM were observed in CP-treated cells. CP and its polyphenolic active components (primarily caffeic acid phenethyl ester and quercetin) had strong inhibitive effects against NF-κB activation and increased the transcriptional activity of Nrf2-ARE. These findings suggest that propolis may be valuable in the control of bovine mastitis.

No MeSH data available.


Related in: MedlinePlus