Research On Holstein-bovine Mammary Epithelial Cell Inflammation Reaction Of Flos Lonicerae And Fructus Forsythia

This paper used6extracts (Flos Lonicerae decoction, Chlorogenic acid, Flos Lonicerae flavonoids,Fructus Forsythiae decoction, Forsythin, Forsythia flavonoids) to act on inflammation response ofHolstein-bovine mammary epithelial cells to explore the mechanism of Flos Lonicerae and FructusForsythiae’s protection for mouse infected by Escherichia coli and picked out the most effectiveingredient.It used tissues explant to culture bovine mammary epithelial cells and purified the cells by trypsindigesting from time interval.80%of the plate would covered with cells in8-10days, then it could obtainpurified mammary epithelial cells by trypsin digesting for2-3times. It was identified that the tissuespecific expression of cytokeratin18genes was positive in mammary epithelial cells byimmunofluorescence cytochemical staining.The filtrate of Escherichia coli culture medium as inflammatory activator was made to produceinflammatory model of epithelial cells. The extracts was added on inflammatory model by each effectiveconcentration. It was to evaluate the protection for the cells of all the extracts by observing the cellmorphology and using MTT method to determine the absorbance of each group. The Flos Loniceraeflavonoids and Forsythia flavonoids both caused damage to cells, almost all of the epithelial cells werebroken to fragment even at the concentration of0.0025g/L.Flos Lonicerae decoction and Chlorogenic acid had the same effective concentration, they both hadsignificant protection for cells at the concentration of1g/L and0.5g/L, and Flos Lonicerae decoction’sprotection was better than Chlorogenic acid. Fructus Forsythiae decoction and Forsythin had the sameeffective concentration, they both had significant protection for cells at the concentration of0.25g/L and0.13g/L, and Fructus Forsythiae decoction was better than Forsythin, Flos Lonicerae decoction (0.5g/L)and Fructus Forsythiae decoction (0.13g/L) both could promote cells proliferation.Elisa was employed to determine the expression level of Interferon-γ(IFN-γ)、Interleukin-4(IL-4) andInterleukin-10(IL-10) in serum of mouse infected by Escherichia coli, than the ratio of IFN-γ/IL-4andIFN-γ/IL-10was compared to explore the mechanism of Flos Lonicerae decoction and Fructus Forsythiaedecoction’s protection for mouse. With the result that the infected mouse death rate was above93%, whilethe death rate descended to be about15%by the curing of Flos Lonicerae decoction and FructusForsythiae decoction.Compared with Normal group, Model group and Cure group of Flos Lonicerae and FructusForsythiae both experienced a remarkable rise in both expression and ratio of IFN-γ、IL-4(P<0.01)in serum;Expression of IFN-γ and ratio of IFN-γ/IL-4had a upward trend, IL-4expression saw a ratio downwardtrend; Expression of IFN-γ and IL-4and ratio of IFN-γ/IL-10in Fructus Forsythiae Cure groupexperienced a downward trend, otherwise ratio of IFN-γ/IL-4ascended. Secretion of Th1/Th2cytokines inrats of Model group increased, and balance of Th1/Th2feathers drifting to Th1.