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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 propolis on mastitis pathogen-induced cell viability decreases and cell apoptosis in MAC-T cells. (a) MAC-T cells were treated with propolis and/or various mastitis pathogens, 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 (Escherichia coli and Staphylococcus aureus, 107 particles/mL) with indicated concentrations of propolis for 24 h. The CCK-8 assay was performed to determine cell viability. ##P < 0.01 and ###P < 0.001 significantly different from untreated cells. ∗P < 0.05 and ∗∗P < 0.01 significantly different from mastitis pathogens-treated cells. (b) After being pretreated with or without Chinese propolis (15 μg/mL) for 1 h, MAC-T cells were challenged with various mastitis pathogens for 24 h. Cell apoptosis was analyzed by flow cytometry analysis using annexin V-FITC and PI staining. The data are expressed as the mean ± SD (n = 3). ∗P < 0.05 and ∗∗P < 0.01.
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fig1: Effects of propolis on mastitis pathogen-induced cell viability decreases and cell apoptosis in MAC-T cells. (a) MAC-T cells were treated with propolis and/or various mastitis pathogens, 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 (Escherichia coli and Staphylococcus aureus, 107 particles/mL) with indicated concentrations of propolis for 24 h. The CCK-8 assay was performed to determine cell viability. ##P < 0.01 and ###P < 0.001 significantly different from untreated cells. ∗P < 0.05 and ∗∗P < 0.01 significantly different from mastitis pathogens-treated cells. (b) After being pretreated with or without Chinese propolis (15 μg/mL) for 1 h, MAC-T cells were challenged with various mastitis pathogens for 24 h. Cell apoptosis was analyzed by flow cytometry analysis using annexin V-FITC and PI staining. The data are expressed as the mean ± SD (n = 3). ∗P < 0.05 and ∗∗P < 0.01.

Mentions: As shown in Figure 1(a), not all of these stimuli caused cell viability decreases in MAC-T cells. Only LPS and heat-killed E. coli and S. aureus, but not TNF-α and LTA stimulation, lead to significant cell viability losses (15% to 52%, P = 0.0009, 0.0041, and 0.001 for LPS, E. coli, and S. aureus, resp.). Heat-killed bacterial strains caused more serious cell viability losses than LPS.


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 propolis on mastitis pathogen-induced cell viability decreases and cell apoptosis in MAC-T cells. (a) MAC-T cells were treated with propolis and/or various mastitis pathogens, 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 (Escherichia coli and Staphylococcus aureus, 107 particles/mL) with indicated concentrations of propolis for 24 h. The CCK-8 assay was performed to determine cell viability. ##P < 0.01 and ###P < 0.001 significantly different from untreated cells. ∗P < 0.05 and ∗∗P < 0.01 significantly different from mastitis pathogens-treated cells. (b) After being pretreated with or without Chinese propolis (15 μg/mL) for 1 h, MAC-T cells were challenged with various mastitis pathogens for 24 h. Cell apoptosis was analyzed by flow cytometry analysis using annexin V-FITC and PI staining. The data are expressed as the mean ± SD (n = 3). ∗P < 0.05 and ∗∗P < 0.01.
© Copyright Policy - open-access
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC4940570&req=5

fig1: Effects of propolis on mastitis pathogen-induced cell viability decreases and cell apoptosis in MAC-T cells. (a) MAC-T cells were treated with propolis and/or various mastitis pathogens, 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 (Escherichia coli and Staphylococcus aureus, 107 particles/mL) with indicated concentrations of propolis for 24 h. The CCK-8 assay was performed to determine cell viability. ##P < 0.01 and ###P < 0.001 significantly different from untreated cells. ∗P < 0.05 and ∗∗P < 0.01 significantly different from mastitis pathogens-treated cells. (b) After being pretreated with or without Chinese propolis (15 μg/mL) for 1 h, MAC-T cells were challenged with various mastitis pathogens for 24 h. Cell apoptosis was analyzed by flow cytometry analysis using annexin V-FITC and PI staining. The data are expressed as the mean ± SD (n = 3). ∗P < 0.05 and ∗∗P < 0.01.
Mentions: As shown in Figure 1(a), not all of these stimuli caused cell viability decreases in MAC-T cells. Only LPS and heat-killed E. coli and S. aureus, but not TNF-α and LTA stimulation, lead to significant cell viability losses (15% to 52%, P = 0.0009, 0.0041, and 0.001 for LPS, E. coli, and S. aureus, resp.). Heat-killed bacterial strains caused more serious cell viability losses than LPS.

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