| To explore the total flavonoids of humulus scandens effects on mammary gland epithelial cells related genes expression.The cow mammary gland epithelial cells as cell model,The living rate of the cells and growth detected by the MTT method,flavonoid solution with different concentration of(0,100,200,400,600,800 mg/L)to stimulate the cow mammary gland epithelial cells 12 h,again with flavonoids solution of the same concentration(400 mg / L)on the cow mammary gland epithelial cells at different times(0 h,1 h,4 h,9 h,12 h,16 h,24 h),and respectively extracted the total RNA of cells,using the fluorescent quantitative PCR method to determine the expression of PRL,GHR,beta-CN,IL-6,IL-8,TNF-alpha and IFN-gamma that are lactation-related genes and to study.Coming to the following results:1)At 4 d,the number of the surviving cells was maximum,after 4 d,the number of the surviving cells presented a downward trend,conforming to the general law of cell growth determined by MTT method.2)Flavonoid solution of the different concentration to stimulate the cow mammary gland epithelial cells,after 12 h,the flavonoids concentration for 100-800mg/L decreased the expression of IL-6 and the difference was significant,but the expression quantity in 800 mg/L was minimum;The expression of IL-8 quantity in800 mg/L felled too,but the difference was not significant;The flavonoids concentration of 100-600 mg/L,the expression quantity of TNF-alpha were lower,but the expression quantity in 200 mg/L was minimum;The expression of IFN-gamma quantity in flavonoids concentration for 200-400 mg/L were both lower,and the expression quantity in the 200 mg/L was minimum.3)Flavonoid solution of the different concentration to stimulate the cow mammary gland epithelial cells,after 12 h,the concentration of flavonoids in 200mg/L,it promoted the expression of GHR,but no significant difference;In 100-800mg/L,the expression quantity of beta-CN had the effect of increase,the flavonoid concentration for 200-600 mg/L significantly increased the expression of beta-CN,but in the concentration of flavonoids of 400 mg/L,the expression quantity of beta –CN was the highest.4)Flavonoid solution of the same concentration(400 mg/L)to stimulate mammary gland epithelial cells in different times,acting on cells for 4 to 9 h,the expression quantity of IL-6 decreased,but acting on cells for 9 h,the expression quantity significantly reduced;For 1 h,the expression quantity of IL-8 decreased,the difference was significant;For 1-24 h,the expression quantity of TNF-alpha were lower,but at the time of 12 h,its expression quantity decreased obviously;For1-24 h,the expression quantity of IFN-gamma were lower,but for 12 h,the expression quantity significantly reduced.5)Flavonoid solution of the same concentration(400 mg/L)to stimulate mammary gland epithelial cells in different times,acting on cells 4 to 9 h,it played a role in promoting the expression quantity of GHR,but for 4 h,the expression quantity significantly increased;Acting on cells 4 to 12 h,it also played a role in promoting the expression quantity of beta-CN and the difference was significant,but at the time of 12 h,the expression quantity was highest.This research gets the following conclusion:1)To cultivate the cow mammary gland epithelial cells,at the time of 4 d,the living rate of the cells was strongest.2)Flavonoids of humulus scandens has something to do with mammary gland epithelial cells related to the expression of inflammation factor ?its dose and action time.The concentration of flavonoids for 800 mg/L,acting on the mammary gland epithelial cells for 9 h,the expression quantity of inflammation factor IL-6significantly reduced;The concentration of flavonoids for 200 mg/L,acting on mammary gland epithelial cells for 12 h,the expression quantity of TNF-alpha and IFN-gamma significantly reduced.It maybe play a role on the inhibition of the mammary gland cells related to the expression of inflammation genes.3)To cultivate the cow mammary gland epithelial cells,at the time of 4 d,the living rate of the cells was strongest.That is,at the moment,the cells had one of the most research value.Flavonoids of humulus scandens has something to do with mammary gland epithelial cells related to the expression of lactation gene ?its dose and action time,The concentration of flavonoids for 400 mg/L,acting on themammary gland epithelial cells for 12 h,it had a positive effect on the expression quantity of beta-CN;The concentration of flavonoids for 400 mg/L,acting on mammary gland epithelial cells for 4 h,it promoted the expression of GHR.These show that it plays an important role on growth and development of mammary gland cells. |
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