The Protective Effect And Mechanisms Of Cepharanthine On LPS-induced Mastitis In Mice

Mastitis, defined as the inflammation of the mammary gland, is a major diseasethat affects the dairy industry. A number of organisms, including bacteria,mycoplasma, yeasts, and algae, are involved in the disease. Mastitis decreases milkproduction and reduces milk quality, which financially impacts dairy producers andaffects the quality of milk available to the consumer. Currently, conventional methodsare unable to prevent the mammary infection. Therefore, the development of noveltherapies for mastitis is urgently needed.Cepharanthine (CEP), a biscoclaurine alkaloid isolated from Stephaniacepharantha Hayata, has been reported to have a number of pharmacologicalproperties and is widely used to treat many acute and chronic diseases without seriousside effects. LPS is an important component of the outer membrane of gram-negativebacteria and has been reported to be an important virulence factor that can be used toestablish animal models of mastitis as tools to study the disease. In the present study,the purpose of this study was to assess the preventive effects of CEP on theLPS-induced mouse mastitis and to further elucidate its potential anti-inflammatorymechanism.Firstly, we observed the effects of cepharanthine on LPS-induced mastitis inmice. Five to seven days after giving birth, the lactating females were randomLydivided into six groups as follows: a blank control group, an LPS group, threetreatment groups, and a DEX (5mg/kg) group. The treatment groups wereadministered with2.5,5, and10mg/kg CEP, respectively, at1h before and12h afterthe LPS infusion, and the DEX group was considered to be the positive control. Theblank control group and the LPS group were treated with an equal volume of distilledwater i.p. The results showed that CEP significantly attenuated the infiltration ofneutrophils, suppresses myeloperoxidase activity, and reduced the levels of TNF-α,IL-1β, and IL-6in LPS-induced mouse mastitis. Furthermore, CEP inhibited thephosphorylation of NF-κB p65subunit and the degradation of its inhibitor IκBα. All the results suggest that CEP exerts potent anti-inflammatory effects on LPS-inducedmouse mastitis.Secondly, to further confirm the protective effect of CEP on LPS-induced mousemastitis, we isolated the mouse mammary epithelial cells from the mammary gland ofmice by primary culture to investigate the antiinflammatory effect of CEP. Thecytotoxicity of CEP on mouse mammary epithelial cells was determined by MTT kit,TNF-α and IL-6levels were measured by ELISA, and NF-κB and Akt signalpathways were analyzed by western blot. The results showed CEP had no cytotoxicityon mouse mammary epithelial cells, and CEP decreased the levels of TNF-α and IL-6.Moreover, CEP inhibited the activation of NF-κB and Akt pathways.In conlusion, we found that CEP protects mice from LPS-induced mastitis by itspotent anti-inflammatory property, and its potential mechanism is associated withPI3K/Akt/NF-κB pathway.