原花青素和茶多酚对兔精子获能的影响

Effects of proanthocyanidins and tea polyphenols on capacitation of rabbit sperm

为探究原花青素和茶多酚对新西兰大白兔精子获能效果及其获能相关基因在获能前后表达差异的影响,在获能液中加入不同浓度(分别为0、0.5、1.0、1.5、2.0mg·mL^-1)的原花青素或茶多酚,与精液悬浮培养,以孕酮诱导顶体反应,并采用实时荧光定量PCR技术检测获能相关基因--蛋白激酶A催化亚单位β(PRKACB)和酸性磷酸酶6(ACP6)在获能前后的表达差异。结果表明:兔精子获能液中原花青素的最适添加量为1-0mg·mL^-1,在此浓度下精子顶体反应率显著高于其他处理,以对照组获能后相关基因ACP6和PRKACB的表达量为1,则原花青素最佳处理组ACP6和PRKACE的相对表达量分别为1.41和3.12,显著高于对照组;茶多酚的最适添加量为1.0mg·mL^-1,在此浓度下精子顶体反应率显著高于其他处理组,以对照组获能后相关基因ACP6和PRKACB的表达量为1,则茶多酚最佳处理组ACP6和PRKACB的相对表达量分别为1.81和1.88,显著高于对照组。综合而言,兔精子获能液中原花青素或茶多酚的适宜添加量均为1.0mg·mL^-1时,在此浓度下,精子顶体反应率较高,ACP和PKA活性增强。

Different concentrations of proanthocyanidins (OPC)(0,0.5, 1.0, 1.5,2.0mg·mL^-1) or tea polyphenols (TPP)(0, 0.5, 1.0, 1.5, 2.0 mg·mL^-1) was added into rabbit sperm to explore the capacitation reaction and the differential expression of key genes related to acrosome reaction before and after capacitation. The capacitation fluid including different concentrations of OPC or TPP was cultured together with sperm suspension, and induced acrosome reaction w让h progesterone. The differential expression of protein kinase A catalyzed subunit β(PRKACB) and acid phosphatase 6 (ACP6) were evaluated before and after capacitation by real-time fluorescent quantitative PCR. The results manifested that the optimum addition of proanthocyanidins at the concentration of 1.0mg·mL^-1 had a significantly higher acrosome reaction in sperm than that in other treatment groups (P<0. 05). The relative expression of ACP6 and PRKACB was 1 in the blank group and the relative expression of ACP6 and PRKACB was 1. 41 and 3. 12 in the procyanidin-optimized treatment group respectively, significantly higher than the control group (P<0.05). The optimum addition of tea polyphenols at the concentration of 1.0 mg·mL^-1 had a significantly higher acrosome reaction in sperm than that in other treatment groups (P<0.05). The relative expression of ACP6 and PRKACB was 1 in the blank group and the relative expression of ACPQ and PRKACB was 1. 41 and 3. 12 in the tea polyphenol-optimized treatment group respectively, significantly higher than the control group (P<0. 05). This experiment indicated that when the concentration of procyani- dins or tea polyphenols was 1. 0 mg·mL^-1 in capacitation fluid of rabbit sperm, the incidence of sperm acrosome was higher, and the activity of ACP and PKA was enhanced.