褪黑素对牛卵巢颗粒细胞发育相关基因表达的作用

Role of melatonin on expression of genes related to ovarian granulosa cell development

目的:探究褪黑素对卵巢颗粒细胞发育相关基因表达的影响。方法:不同浓度的褪黑素(0、10^(-3)、10^(-5)、10^(-7)、10^(-9)、10^(-11)mol/L)分别处理颗粒细胞24、48、72 h后提取细胞总RNA,采用Real-time PCR检测抗氧化(GPX4、SOD1)、发育(EGFR、DNMT1a、FSHR)和TGFβ家族(BMP6)相关基因的表达。结果:不同浓度褪黑素处理24、48、72 h对抗氧化(GPX4、SOD1)、发育(EGFR、DNMT1a、FSHR)和TGFβ家族(BMP6)相关基因的表达均有促进或抑制作用(P<0.05),高浓度褪黑素(10^(-3)mol/L)在48 h对GPX4、SOD1的促进效果最显著(P<0.05);高浓度褪黑素(10^(-3)mol/L)不影响EGFR、DNMT1a的表达,其他浓度褪黑素均促进EGFR、DNMT1a的表达(P<0.05);低浓度褪黑素(10^(-11)mol/L)显著抑制FSHR的表达(P<0.05);褪黑素对于BMP6的表达在24 h为促进作用(P<0.05),48 h时抑制其表达(P<0.05),72 h时高浓度褪黑素(10^(-3)mol/L)和低浓度褪黑素(10^(-11)mol/L)无显著作用,其他浓度为促进作用(P<0.05)。结论:褪黑素可以通过促进抗氧化相关基因(GPX4、SOD1)、发育相关基因(EGFR、DNMT1a)和TGFβ家族(BMP6)相关基因的表达,抑制FSHR的表达来保护颗粒细胞,抑制其凋亡。

Objective:To investigate the effect of melatonin on the expression of development-related genes in ovarian granulosa cells.Methods:Total cellular RNA was extracted after treating granulosa cells with different concentrations of melatonin(0,10^(-3),10^(-5),10^(-7),10^(-9),and10^(-11)mol/L)for 24,48,72 h,respectively,and the expression of genes related to antioxidant(GPX 4,SOD 1),developmental(EGFR,DNMT 1 a,and FSHR),and TGFβfamily(BMP 6)was detected by real-time PCR.Results:Different concentrations of melatonin promoted or inhibited the expression of antioxidant(GPX 4,SOD 1),developmental(EGFR,DNMT 1 a,FSHR),and TGFβfamily(BMP 6)-related genes at 24,48,72 h(P<0.05),and a high concentration of melatonin(10^(-3)mol/L)had the most significant(P<0.05)promotional effect on GPX 4,SOD 1,and SOD 1 at 48 h.The high concentration of melatonin(10^(-3)mol/L)had the most significant(P<0.05)promotional effect on GPX 4 and SOD 1 at 48 h(P<0.05).SOD 1 at 48 h.High concentration melatonin(10^(-3)mol/L)had the most significant effect on GPX 4 and SOD 1(P<0.05).High concentration melatonin(10^(-3)mol/L)did not affect the expression of EGFR and DNMT 1 a,and all other concentrations of melatonin promoted the expression of EGFR and DNMT 1 a(P<0.05),and low concentration melatonin(10^(-11) mol/L)significantly inhibited the expression of FSHR(P<0.05),and the effect of melatonin on BMP 6 expression at 24 h was promotional(P<0.05).Expression of BMP 6 at 24 h(P<0.05)and inhibited its expression at 48 h(P<0.05).At 72 h,the high concentration of melatonin(10^(-3)mol/L)and the low concentration of melatonin(10^(-11) mol/L)did not have any significant effect,and the other concentrations had a promotional effect(P<0.05).Conclusion:Melatonin can protect granulosa cells and inhibit their apoptosis by promoting the expression of antioxidant-related genes(GPX 4,SOD 1),development-related genes(EGFR,DNMT 1 a),and TGFβfamily(BMP 6)-related genes,and inhibiting the expression of FSHR.