褪黑素对牦牛黄体细胞生长及功能影响的研究

本试验旨在探究褪黑素(melatonin,MT)对牦牛黄体细胞生长、抗氧化性及功能的影响.本试验以健康牦牛二代黄体细胞为研究对象,比较了不同浓度MT(0(Control组)、25、125、250、500 pg·mL-1)对牦牛黄体细胞生长和功能的影响.通过CCK-8法检测黄体细胞的生长情况,并采用qRT-PCR检测增殖...

Research on the Effect of Melatonin on the Growth and Function of Yak Luteal Cells

This experiment was intended to research the effect of melatonin(MT)on the growth,antioxidant activity and function in yak luteal cells.The 2nd-passage luteal cells from healthy yak were used as research object in this experiment,and the effect of different concentrations(0(Control),25,125,250,and 500 pg·mL^-1)of melatonin on the growth and function in yak luteal cells were compared.Cell proliferation and viability were evaluated by CCK-8 assay.qRT-PCR was conducted to analyse the mRNA expression levels of several genes,including proliferation-related gene PCNA,apoptosis-related genes BCL-2,BAX,FAS,antioxidant-related genes SOD1,SOD2,GPX1 and CAT,as well as progesterone synthesis-related genes HSD3β,STAR and CYP11A1.The levels of ROS and progesterone were determined by ELISA.The results showed that the supplementation of different concentrations of MT could promote luteal cells proliferation and accelerate them into the plateau phase.MT could promote the mRNA expression of cell-proliferated gene PCNA and anti-apoptosis-related gene BCL-2,and repress the expression of apoptosis-related genes BAX,FAS.MT could reduce the ROS level and promote progesterone secretion significantly in luteal cells,moreover,the above effects were most obvious when the MT concentration was 250 pg·mL^-1.At this concentration,the mRNA expression levels of antioxidant-related genes SOD2,GPX1,CAT,and progesterone synthesis-related genes STAR and CYP11A1 significantly up-regulated.Compared with MT treatment alone,the co-treatment with MT and MT receptor inhibitor Luzindole could significantly increase the ROS level and decrease the expression level of SOD2,GPX1 and CAT.The level of progesterone and the expression of STAR and CYP11A1 decreased significantly.In conclusion,MT promotes yak luteal cells proliferation by up-regulating the mRNA expression level of PCNA and BCL-2,and repressing the expression level of BAX and FAS.MT may regulate the ROS level and progesterone level through its receptor.This study provides theoretical basis for the application of MT in the treatment of reproductive diseases caused by corpus luteum abnormality and the improvement in the reproductive performance of yak.