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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

Chinese propolis and its major bioactive compounds suppressed mastitis pathogen-induced NF-κB activation and induces ARE transcriptional activity. (a) Effects of CP on mastitis pathogen-induced NF-κB promoter activation in HEK-293T cells. Cells were pretreated with 20 μg/mL CP and identified isolated active compounds from propolis (caffeic acid, 50 μM; CAPE, 5 μM; chrysin, 50 μM; ferulic acid, 50 μM; galangin, 25 μM; kaempferol, 50 μM; pinocembrin, 50 μM; and quercetin, 50 μM) for 1 h and then stimulated with TNF-α (25 ng/mL) for another 12 h. ###P < 0.001 compared to the vehicle control; ∗P < 0.05 and ∗∗∗P < 0.001 compared to TNF-α control. (b) Effects of propolis on mastitis pathogens-induced ARE promoter activation in HEK-293T cells. Cells were treated with CP or identified isolated active compounds. tBHQ (15 μM) treatment for 7 h was used as ARE positive control. ∗P < 0.05 and ∗∗∗P < 0.001 compared to the vehicle control. The data represent the mean ± SD of four independent experiments.
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fig5: Chinese propolis and its major bioactive compounds suppressed mastitis pathogen-induced NF-κB activation and induces ARE transcriptional activity. (a) Effects of CP on mastitis pathogen-induced NF-κB promoter activation in HEK-293T cells. Cells were pretreated with 20 μg/mL CP and identified isolated active compounds from propolis (caffeic acid, 50 μM; CAPE, 5 μM; chrysin, 50 μM; ferulic acid, 50 μM; galangin, 25 μM; kaempferol, 50 μM; pinocembrin, 50 μM; and quercetin, 50 μM) for 1 h and then stimulated with TNF-α (25 ng/mL) for another 12 h. ###P < 0.001 compared to the vehicle control; ∗P < 0.05 and ∗∗∗P < 0.001 compared to TNF-α control. (b) Effects of propolis on mastitis pathogens-induced ARE promoter activation in HEK-293T cells. Cells were treated with CP or identified isolated active compounds. tBHQ (15 μM) treatment for 7 h was used as ARE positive control. ∗P < 0.05 and ∗∗∗P < 0.001 compared to the vehicle control. The data represent the mean ± SD of four independent experiments.

Mentions: To determine the effects of CP as well as its purified compounds on NF-κB and ARE transcriptional activity, luciferase reporter assays were applied in HEK-293T cells. As shown in Figure 5, NF-κB activity was significantly decreased by approximately 70% (P < 0.001). The luciferase activity derived from the ARE promoter was consistently increased by approximately 4.7-fold after treating with 20 μg/mL CP (Figure 5(b), P < 0.001). Moreover, significant decreases in NF-κB activity were found in cells treated with CAPE (5 μM, 45%, P < 0.001), quercetin (50 μM, 73%, P < 0.001) caffeic acid (50 μM, 12%, P < 0.05), and ferulic acid (50 μM, 11%, P < 0.05) (Figure 5(a)). In parallel, the luciferase activity derived from the ARE promoter was consistently increased after treatment with CAPE (5 μM, 2.5-fold, P < 0.001), quercetin (50 μM, 5.1-fold, P < 0.01), kaempferol (50 μM, 3.6-fold, P < 0.001), and pinocembrin (50 μM, 2.9-fold, P < 0.01).


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)

Chinese propolis and its major bioactive compounds suppressed mastitis pathogen-induced NF-κB activation and induces ARE transcriptional activity. (a) Effects of CP on mastitis pathogen-induced NF-κB promoter activation in HEK-293T cells. Cells were pretreated with 20 μg/mL CP and identified isolated active compounds from propolis (caffeic acid, 50 μM; CAPE, 5 μM; chrysin, 50 μM; ferulic acid, 50 μM; galangin, 25 μM; kaempferol, 50 μM; pinocembrin, 50 μM; and quercetin, 50 μM) for 1 h and then stimulated with TNF-α (25 ng/mL) for another 12 h. ###P < 0.001 compared to the vehicle control; ∗P < 0.05 and ∗∗∗P < 0.001 compared to TNF-α control. (b) Effects of propolis on mastitis pathogens-induced ARE promoter activation in HEK-293T cells. Cells were treated with CP or identified isolated active compounds. tBHQ (15 μM) treatment for 7 h was used as ARE positive control. ∗P < 0.05 and ∗∗∗P < 0.001 compared to the vehicle control. The data represent the mean ± SD of four independent experiments.
© Copyright Policy - open-access
Related In: Results  -  Collection

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fig5: Chinese propolis and its major bioactive compounds suppressed mastitis pathogen-induced NF-κB activation and induces ARE transcriptional activity. (a) Effects of CP on mastitis pathogen-induced NF-κB promoter activation in HEK-293T cells. Cells were pretreated with 20 μg/mL CP and identified isolated active compounds from propolis (caffeic acid, 50 μM; CAPE, 5 μM; chrysin, 50 μM; ferulic acid, 50 μM; galangin, 25 μM; kaempferol, 50 μM; pinocembrin, 50 μM; and quercetin, 50 μM) for 1 h and then stimulated with TNF-α (25 ng/mL) for another 12 h. ###P < 0.001 compared to the vehicle control; ∗P < 0.05 and ∗∗∗P < 0.001 compared to TNF-α control. (b) Effects of propolis on mastitis pathogens-induced ARE promoter activation in HEK-293T cells. Cells were treated with CP or identified isolated active compounds. tBHQ (15 μM) treatment for 7 h was used as ARE positive control. ∗P < 0.05 and ∗∗∗P < 0.001 compared to the vehicle control. The data represent the mean ± SD of four independent experiments.
Mentions: To determine the effects of CP as well as its purified compounds on NF-κB and ARE transcriptional activity, luciferase reporter assays were applied in HEK-293T cells. As shown in Figure 5, NF-κB activity was significantly decreased by approximately 70% (P < 0.001). The luciferase activity derived from the ARE promoter was consistently increased by approximately 4.7-fold after treating with 20 μg/mL CP (Figure 5(b), P < 0.001). Moreover, significant decreases in NF-κB activity were found in cells treated with CAPE (5 μM, 45%, P < 0.001), quercetin (50 μM, 73%, P < 0.001) caffeic acid (50 μM, 12%, P < 0.05), and ferulic acid (50 μM, 11%, P < 0.05) (Figure 5(a)). In parallel, the luciferase activity derived from the ARE promoter was consistently increased after treatment with CAPE (5 μM, 2.5-fold, P < 0.001), quercetin (50 μM, 5.1-fold, P < 0.01), kaempferol (50 μM, 3.6-fold, P < 0.001), and pinocembrin (50 μM, 2.9-fold, P < 0.01).

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