Analysis of the main chemical components of supercritical extract residues of propolis and the evaluation of its effect on the prevention and treatment of dairy cow mastitis
In order to explore the chemical components of supercritical extract residues of propolis and supercritical ethanol extract residues of propolis(alcohol-extraction supercritical residues of propolis),and the effect of the two extract residues on the prevention and treatment of dairy cow mastitis,in the test,referring to the national standard,the content of basic nutrients and mineral elements in the two extract residues were determined.The contents of total phenols and total flavonoids were determined by the flulinol method and the sodium nitrite-aluminum nitrate method[NaNO2-Al(NO3)3]method,respectively.At the same time,six major polyphenol components were identified by high-performance liquid chromatography.Furthermore,lipopolysaccharide was used to induce inflammation in dairy cow mammary epithelial cells(MAC-T).The CCK-8 method was used to detect the effects of supercritical extract residues of propolis and alcohol-extraction supercritical residues of propolis on the viability of MAC-T cells.Real-time quantitative PCR was used to evaluate the effect of supercritical extract residues of propolis and alcohol-extraction supercritical residues of propolis on the mRNA relative expression of MAC-T inflammatory cytokines,such as tumour-necrosis factor-α(TNF-α),interlukin-1β(IL-1β)and interlukin-6(IL-6).The results showed that there was no significant difference in the nutrient contents of the two extract residues,but the fat content of supercritical extract residues of propolis was significantly lower than that of alcohol-extraction supercritical residues of propolis(P<0.05).The contents of total phenols and total flavonoids in supercritical extract residues of propolis were 178.21,514.60 mg/g,respectively,which were significantly higher than those of alcohol-extraction supercritical residues of propolis(P<0.05).The total contents of the six polyphenols were higher than those of alcohol-extraction supercritical residues of propolis.The contents of brevisin,chrysin and galangin in supercritical extract residues of propolis were significantly higher than those in alcohol-extraction supercritical residues of propolis(P<0.05),while there was no significant difference in the contents of caffeic acid and pinein(P>0.05).The contents of calcium,magnesium and potassium in supercritical extract residues of propolis were significantly lower than those in alcohol-extraction supercritical residues of propolis(P<0.05).The contents of sodium,aluminum and manganese were significantly higher than those of alcohol-extraction supercritical residues of propolis(P<0.05),and there was no significant difference in the iron content between the two extract residues(P>0.05).25-200 μg/mL supercritical extract residues of propolis and alcohol-extraction supercritical residues of propolis had no significant cytotoxicity to MAC-T cells.In addition,supercritical extract residues of propolis and alcohol-extraction supercritical residues of propolis significantly reduced the mRNA relative expression of inflammatory cytokines TNF-α,IL-1β and IL-6 in MAC-T cells(P<0.01).The results indicated that supercritical extract residues of propolis and alcohol-extraction supercritical residues of propolis still contained some polyphenolic compounds such as flavonoids and phenolic acids,and could prevent and treat mastitis by reducing the level of proinflammatory factors inflammation in dairy cows.
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