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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

Effects of CP treatment on proinflammatory responses following mastitis pathogen challenges in MAC-T cells. Quantitative PCR analysis of inflammatory cytokine genes, IL-6 (a), IL-8 (b), and TNF-α (c), showing gene expressions after 6 h incubation of MAC-T cells with each different mastitis pathogen, including proinflammatory cytokine (TNF-α 25 ng/mL), bacterial cell wall components (LPS, 1 μg/mL; LTA, 10 μg/mL), and heat-killed mastitis strains (E. coli and S. aureus, 107 particles/mL). Ct values of target genes were normalized to the value of β-actin and relative gene expressions in TNF-α control group were arbitrarily set to one. The data are shown as mean ± SD from three independent experiments and were analyzed by one-way ANOVA with the Student-Newman-Keuls method. The means with different superscripts are significantly different (P < 0.05).
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fig2: Effects of CP treatment on proinflammatory responses following mastitis pathogen challenges in MAC-T cells. Quantitative PCR analysis of inflammatory cytokine genes, IL-6 (a), IL-8 (b), and TNF-α (c), showing gene expressions after 6 h incubation of MAC-T cells with each different mastitis pathogen, including proinflammatory cytokine (TNF-α 25 ng/mL), bacterial cell wall components (LPS, 1 μg/mL; LTA, 10 μg/mL), and heat-killed mastitis strains (E. coli and S. aureus, 107 particles/mL). Ct values of target genes were normalized to the value of β-actin and relative gene expressions in TNF-α control group were arbitrarily set to one. The data are shown as mean ± SD from three independent experiments and were analyzed by one-way ANOVA with the Student-Newman-Keuls method. The means with different superscripts are significantly different (P < 0.05).

Mentions: As shown in Figure 2, all of these mastitis pathogens, except for LTA, lead to significant increases of IL-6 and IL-8 (P < 0.05). LTA and heat-killed S. aureus stimulation also failed to induce expression of TNF-α compared with uninfected cells (P > 0.05). On the contrary, heat-killed E. coli lead to the strongest inductive effects compared to other mastitis stimuli. CP-pretreated MAC-T cells have much slighter changes of IL-6 and TNF-α mRNA compared to their corresponding stimuli controls. Surprisingly, CP lead to ~8-fold changes of IL-8, significantly higher than TNF-α, LPS, and heat-killed E. coli stimuli (P < 0.05).


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)

Effects of CP treatment on proinflammatory responses following mastitis pathogen challenges in MAC-T cells. Quantitative PCR analysis of inflammatory cytokine genes, IL-6 (a), IL-8 (b), and TNF-α (c), showing gene expressions after 6 h incubation of MAC-T cells with each different mastitis pathogen, including proinflammatory cytokine (TNF-α 25 ng/mL), bacterial cell wall components (LPS, 1 μg/mL; LTA, 10 μg/mL), and heat-killed mastitis strains (E. coli and S. aureus, 107 particles/mL). Ct values of target genes were normalized to the value of β-actin and relative gene expressions in TNF-α control group were arbitrarily set to one. The data are shown as mean ± SD from three independent experiments and were analyzed by one-way ANOVA with the Student-Newman-Keuls method. The means with different superscripts are significantly different (P < 0.05).
© Copyright Policy - open-access
Related In: Results  -  Collection

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getmorefigures.php?uid=PMC4940570&req=5

fig2: Effects of CP treatment on proinflammatory responses following mastitis pathogen challenges in MAC-T cells. Quantitative PCR analysis of inflammatory cytokine genes, IL-6 (a), IL-8 (b), and TNF-α (c), showing gene expressions after 6 h incubation of MAC-T cells with each different mastitis pathogen, including proinflammatory cytokine (TNF-α 25 ng/mL), bacterial cell wall components (LPS, 1 μg/mL; LTA, 10 μg/mL), and heat-killed mastitis strains (E. coli and S. aureus, 107 particles/mL). Ct values of target genes were normalized to the value of β-actin and relative gene expressions in TNF-α control group were arbitrarily set to one. The data are shown as mean ± SD from three independent experiments and were analyzed by one-way ANOVA with the Student-Newman-Keuls method. The means with different superscripts are significantly different (P < 0.05).
Mentions: As shown in Figure 2, all of these mastitis pathogens, except for LTA, lead to significant increases of IL-6 and IL-8 (P < 0.05). LTA and heat-killed S. aureus stimulation also failed to induce expression of TNF-α compared with uninfected cells (P > 0.05). On the contrary, heat-killed E. coli lead to the strongest inductive effects compared to other mastitis stimuli. CP-pretreated MAC-T cells have much slighter changes of IL-6 and TNF-α mRNA compared to their corresponding stimuli controls. Surprisingly, CP lead to ~8-fold changes of IL-8, significantly higher than TNF-α, LPS, and heat-killed E. coli stimuli (P < 0.05).

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