苜蓿黄酮对体外培养的奶牛乳腺上皮细胞增殖与抗氧化的影响

本试验旨在研究不同浓度苜蓿黄酮对体外培养奶牛乳腺上皮细胞增殖与抗氧化的影响。试验利用含不同浓度0μg/m L(对照组)、25μg/m L(试验Ⅰ)、50μg/m L(试验Ⅱ)、75μg/m L(试验Ⅲ)、100μg/m L(试验Ⅳ)苜蓿黄酮的培养基进行细胞培养。结果表明,1)细胞培养第3天,各组细胞增殖无显著差异,但第5天试验Ⅱ~Ⅳ组细胞增殖能力极显著高于对照组和试验I组(P0.01)。2)试验Ⅱ~Ⅳ组一氧化氮(NO)水平明显高于对照组与试验Ⅰ组,且差异极显著(P0.01)。试验Ⅰ组、Ⅱ组、Ⅳ组的乳酸脱氢酶(LDH)活性显著低于对照组与试验Ⅲ组,且差异极显著(P0.01),但对照组与试验Ⅲ组差异不显著。3)试验Ⅳ组细胞内过氧化氢酶(CAT)含量极显著高于其他各组(P0.01),而试验Ⅲ组丙二醛(MDA)含量极显著低于其他各组(P0.01);试验Ⅱ和Ⅲ组的谷胱甘肽过氧化物酶(GSH-PX)活性极显著高于其他各组(P0.01),而各处理组细胞的超氧化物歧化酶(SOD)含量差异不显著。4)试验Ⅱ和Ⅲ组细胞p53、Caspase-3、SOCS3和STAT1基因的相对表达量极显著低于对照组(P0.01)。因此,苜蓿黄酮能够促进体外培养奶牛乳腺上皮细胞的增殖,提高细胞抗氧化的能力,抑制细胞凋亡,其中添加浓度为75μg/m L组效果最好。

Proliferation stimulus and antioxidant effect of alfalfa flavonoids on dairy cow mam-mary epithelial cells cultured in vitro

This experiment studied the proliferation stimulus and antioxidant effects of different concentrations of alfalfa flavonoids on cow mammary epithelial cells cultured in vitro .Media with five different concentrations of alfalfa flavonoids were prepared to culture dairy cow mammary epithelial cells in vitro .Concentrations tested were:0 μg/mL (control group),25 μg/mL (trial group Ⅰ),50 μg/mL (trial group Ⅱ),75 μg/mL (trial group Ⅲ),100 μg/mL (trial group Ⅳ).The results showed:1)There was no significant difference in cell pro-liferation between the five concentrations during the first three days of cell culture,but the cell proliferation of trial groups Ⅱ-Ⅳ was significantly higher than for the control group and trial group Ⅰ (P <0.01 )at day5.2)The NO levels of trial groups Ⅱ-Ⅳ were significantly higher than the control group and trial group Ⅰ(P <0.01).Also,LDH activity of groups Ⅰ,Ⅱ,and Ⅳ was significantly lower than the control group and than test group Ⅲ (P <0.01),and the difference between the control group and test group Ⅲ was not signifi-cant.3)Intracellular CAT content of trial group Ⅳ was significantly higher than the other groups (P <0.01). The MAD content of group Ⅲ was significantly lower than the other groups (P <0.01 ).GSH-PX activity of groups Ⅱ and Ⅲ was significantly higher than the other groups (P <0.01).In addition,there was no signifi-cant difference between the five groups in the cell SOD content.4 )The relative expression levels of p53, Caspase-3,SOCS3 and STAT1 genes in groups Ⅱ and Ⅲ were significantly lower than control group (P <0.01).In summary,alfalfa flavonoids can promote the proliferation of bovine mammary epithelial cells cul-tured in vitro ,improve cell antioxidant capacity and inhibit cellular apoptosis.The best effect occurred at a dosage of 75 μg/mL of alfalfa flavonoids.