Effect of chlorogenic acid (CGA) on nuclear factor-κB (NF-κB) activation.

The NF-κB pathway was analyzed in BMECs via Western blot. β-actin was used as a reference control. Dexamethasone (DEX) was used as a positive control. (A) The quantification histogram of p-IκBα protein expression normalized by β-actin; and (B) the quantification histogram of p-p65 protein expression normalized by β-actin. The values presented are the mean ± SE of three independent experiments. **p S1 Raw images NC: normal control; LTA: lipoteichoic acid; LTA+CGA: lipoteichoic acid and chlorogenic acid; DEX: Dexamethasone.</p

Quantitative real-time PCR primer information.

Mastitis is an inflammatory disease caused by microbial infection. Chlorogenic acid (CGA), one of the major phenolic acids in Taraxacum officinale, has natural antioxidant and anti-inflammatory properties in various cell types; however, the effects of CGA on Lipoteichoic acid (LTA)-induced bovine mammary epithelial cells (BMECs) have not been investigated. In this study, the CGA content in T. officinale was determined by High-performance liquid chromatography (HPLC). BMECs were infected with LTA to induce the mastitis model. Different concentrations of CGA were administered after establishing the LTA infection. The results showed that the T. officinale contained CGA 1.36 mg/g. CGA significantly reduced the pro-inflammatory gene and protein expression of TNF-α, IL-6, and IL-1β. In addition, CGA downregulated the NO, TLR2, and NF-κB signaling pathways in LTA-infected bovine mammary epithelial cells. Our results indicate that CGA reduced the expression of TNF-α, IL-6, IL-1β, and TLR2 by inhibiting the phosphorylation of proteins in the NF-κB signaling pathways in a dose-dependent manner. This finding suggests that CGA may be a potential agent for the treatment of mastitis in dairy cows.</div

S1 Raw images -

Mastitis is an inflammatory disease caused by microbial infection. Chlorogenic acid (CGA), one of the major phenolic acids in Taraxacum officinale, has natural antioxidant and anti-inflammatory properties in various cell types; however, the effects of CGA on Lipoteichoic acid (LTA)-induced bovine mammary epithelial cells (BMECs) have not been investigated. In this study, the CGA content in T. officinale was determined by High-performance liquid chromatography (HPLC). BMECs were infected with LTA to induce the mastitis model. Different concentrations of CGA were administered after establishing the LTA infection. The results showed that the T. officinale contained CGA 1.36 mg/g. CGA significantly reduced the pro-inflammatory gene and protein expression of TNF-α, IL-6, and IL-1β. In addition, CGA downregulated the NO, TLR2, and NF-κB signaling pathways in LTA-infected bovine mammary epithelial cells. Our results indicate that CGA reduced the expression of TNF-α, IL-6, IL-1β, and TLR2 by inhibiting the phosphorylation of proteins in the NF-κB signaling pathways in a dose-dependent manner. This finding suggests that CGA may be a potential agent for the treatment of mastitis in dairy cows.</div

The effect of chlorogenic acid (CGA) on the cell viability of bovine mammary epithelial cells (BMECs).

The values presented are the mean ± SE of three independent experiments. ## p < 0.01 vs. control group; * p < 0.05, ** p < 0.01 vs. LTA group. NC: normal control; LTA: lipoteichoic acid; LTA+CGA: lipoteichoic acid and chlorogenic acid.</p

Effect of chlorogenic acid (CGA) on lipoteichoic acid (LTA)-induced protein expression of inflammatory cytokines.

The values presented are the mean ± SE of three independent experiments. ## p < 0.01 vs. control group; * p < 0.05, ** p < 0.01 vs. LTA group. NC: normal control; LTA: lipoteichoic acid; LTA+CGA: lipoteichoic acid and chlorogenic acid.</p

Results of high-performance liquid chromatogram of the chlorogenic acid.

(A) HPLC of the chlorogenic acid standard, and (B) HPLC of the chlorogenic acid in dandelion sample.</p

Effect of chlorogenic acid (CGA) on toll-like receptor 2 (TLR2) expression.

The expression of TLR2 in BMECs was analyzed via Western blot. β-actin was used as a reference control. Dexamethasone (DEX) was used as a positive control. (A) The quantification histogram of TLR2 protein expression normalized by β-actin. The values presented are the mean ± SE of three independent experiments. ** p S1 Raw images. NC: normal control; LTA: lipoteichoic acid; LTA+CGA: lipoteichoic acid and chlorogenic acid; DEX: Dexamethasone.</p

Effect of chlorogenic acid (CGA) on Nitric oxide (NO) production.

The values presented are the mean ± SE of three independent experiments. # p < 0.05 vs. control group; ** p < 0.01 vs. LTA group. NC: normal control; LTA: lipoteichoic acid; LTA+CGA: lipoteichoic acid and chlorogenic acid.</p

Effect of chlorogenic acid (CGA) on lipoteichoic acid (LTA)-induced gene expression of inflammatory cytokines.

The values presented are the mean ± SE of three independent experiments. ## p < 0.01 vs. control group; * p < 0.05, ** p < 0.01 vs. LTA group. NC: normal control; LTA: lipoteichoic acid; LTA+CGA: lipoteichoic acid and chlorogenic acid.</p

Image_1_The Antimicrobial Peptide MPX Can Kill Staphylococcus aureus, Reduce Biofilm Formation, and Effectively Treat Bacterial Skin Infections in Mice.jpeg

Staphylococcus aureus is a common pathogen that can cause pneumonia and a variety of skin diseases. Skin injuries have a high risk of colonization by S. aureus, which increases morbidity and mortality. Due to the emergence of multidrug-resistant strains, antimicrobial peptides are considered to be among the best alternatives to antibiotics due to their unique mechanism of action and other characteristics. MPX is an antibacterial peptide extracted from wasp venom that has antibacterial activity against a variety of bacteria. This study revealed that MPX has good bactericidal activity against S. aureus and that its minimum inhibitory concentration (MIC) is 0.08 μM. MPX (4×MIC) can kill 99.9% of bacteria within 1 h, and MPX has good stability. The research on the bactericidal mechanism found that MPX could destroy the membrane integrity, increase the membrane permeability, change the membrane electromotive force, and cause cellular content leakage, resulting in bactericidal activity. Results from a mouse scratch model experiment results show that MPX can inhibit colonization by S. aureus, which reduces the wound size, decreases inflammation, and promotes wound healing. This study reports the activity of MPX against S. aureus and its mechanism and reveals the ability of MPX to treat S. aureus infection in mice, laying the foundation for the development of new drugs for bacterial infections.</p