Effects Of Glycosides On Proliferation And Casein Gene Expression Of Gost Mammary Gland Epithelial Cells

Tissue culture was used in this study to separate goat mammary epithelial cells. Then curettage law and difference digest were used to purify cells. Effects of four glycosides(daidzin, genistin, icariin and total platycosides) on proliferation and expression of casein αs1and β gene of two or three generations of the cells were studied.1. Culture and identification of goat mammary epithelial cells(GMEC)Through tissue culture method, this test focused on primary cell culture, using curettage law. difference digest and adherent law to purify cells, with0.05percent of tr-ysin and0.04percent of EDTA to digest passage. The cells which could be showed from morphology observation and immunization section test were epithelial cells. The growth of cells in good condition could be got, and the morphology of cells was same.The viability of20th passage cells decreased. Cell apoptosis happeded in25passage cells, and30th passage cells died.2. Effects of Glycosides on proliferation of GMECThe Effects of daidzin, genistin, icariin and total platycosides on proliferation of goat mammary gland epithelial cells were measured by MTT. The results revealed that the ng level of concentration had more effects on cells than the μg level of concentration. And when cells were treated with the four materials for48h, the effects were greater than for24h and72h.The highest poteney was detected in daidzin (10ng/mL) and genistin (10ng/mL and100ng/mL) compared with the control group. All the concentration groups (1,5,10,100and200ng/mL) could significantly improve the proliferation of cells, and there was dose dependence relation.However two groups of total platycosides(1ng/mLand5ng/mL) would inhibit the proliferation of goat mammary gland epithelial cells.3. Effects of Glycosides on the expression of casein αs1and β gene in GMECTo study the effects of daidzin, genistin, icariin and total platycosides on the expression of casein αs1and β gene in GMEC, the second or third passage cells were selected as experimental subjects.The expression of αs1-ceasein and β-casein mRNA in GMEC’s lysate were detected by using Real Time PCR. The results showed that: Compared with the control group, the expression of αs1-casein of the group daidzin (1ng/mL and lOng/mL) were increased, but there were no significantly differences. The expression of αs1-casein of the group genistin (10ng/mL and100ng/mL), icariin (1ng/mL,10ng/mL and100ng/mL) and total platycosides (1ng/mL and100ng/mL) were most significantly inereased. The expression of β-casein of the group daidzin (10ng/mL) was increased, but there were no significantly differences. The expression of β-casein of the group genistin (1ng/mL and10ng/mL), icariin (1ng/mL,10ng/mL and100ng/mL) and total platycosides (1ng/mL and10ng/mL) were most signifieantly inereased. The best concentration of daidzin, genistin, icariin and total platycosides were10ng/mL,10ng/mL,100ng/mL and10ng/mL,100ng/mL, respectively. And when the concentrations of icariin were higher, The expression of αS1-casein and β-casein gene were more significantly inereased.