Berberine-Loaded Composite Phospholipid Ethosome Hydrogels: A Therapeutic for Mastitis via Regulating the NF-κB and PI3K/Akt Signaling Pathway

Simple SummaryMastitis is a common disease in dairy cows, with major pathogens includingStaphylococcus aureusandEscherichia coli. Although berberine has antibacterial and anti-inflammatory properties, it is poorly absorbed after oral administration. We developed a novel berberine-loaded hydrogel that can be applied to the udder, delivering the drug directly to the mammary tissue. Experimental results showed that this hydrogel is stable, safe, and capable of sustained drug release. Tests in a mouse model demonstrated that it effectively reduces bacteria-induced inflammation, protects the mammary barrier, and minimizes cell damage. Its mechanism of action is related to the regulation of intracellular NF-κB and PI3K/Akt signaling pathways. This study suggests that the hydrogel offers a new topical strategy for treating mastitis, solving the problem of poor oral absorption through local application. It holds promise for providing dairy cows with a more convenient and effective treatment option in the future.Mastitis is a common disease in dairy cows, mainly caused by Staphylococcus aureus and Escherichia coli. Berberine (BBR) has antibacterial and anti-inflammatory potential, but its application is limited due to poor oral absorption and difficulty in reaching mammary tissue. To address this, this study developed a BBR-loaded composite ethosome hydrogel (BBR-CEH) to achieve targeted mammary delivery through local transdermal administration. The experimental results showed that BBR-CEH has good chemical stability and biosafety. Subsequently, a mouse mastitis model was established by intraductal injection of 50 µL of bacterial mixture (E. coli:S. aureus= 1:1, each at 1 × 107CFU/mL). The results showed that after BBR-CEH treatment, the mRNA expression ofTNF-α(tumor necrosis factor-alpha),IL-6(interleukin-6), andIL-1β(interleukin-1 beta) was significantly decreased, the mRNA expression ofZO-1(zonula occludens-1),Occludin, andClaudin-4was significantly increased, andBax/Bcl-2(Bcl-2-associated X protein/B-cell lymphoma 2) was significantly reduced (p< 0.01), indicating alleviation of mastitis by reducing inflammation, improving tight junctions, and inhibiting apoptosis. Finally, network pharmacology and in vivo experiments confirmed that its mechanism involves the NF-κB (nuclear factor kappa-B) and PI3K/Akt (phosphoinositide 3-kinase/protein kinase B) pathways. Thus, topical BBR-CEH may represent a promising new strategy for mastitis treatment.