目的探讨β-羟基丁酸(BHB)对SD大鼠神经元细胞缺氧损伤的保护作用及其机制。方法原代培养SD大鼠神经元细胞,不同浓度(2 m M、5 m M、10 m M、20 m M和50 m M)BHB预处理24 h,三气培养箱糖氧剥夺培养2 h,线粒体超氧化物探针检测细胞线粒...目的探讨β-羟基丁酸(BHB)对SD大鼠神经元细胞缺氧损伤的保护作用及其机制。方法原代培养SD大鼠神经元细胞,不同浓度(2 m M、5 m M、10 m M、20 m M和50 m M)BHB预处理24 h,三气培养箱糖氧剥夺培养2 h,线粒体超氧化物探针检测细胞线粒体氧化应激状态,CCK8检测细胞活力变化,QPCR测Na+偶联单羧酸转运体1(SMCT1)、caspae3和Cytochrome C的m RNA表达,Western Blot测SMCT1、caspase3、细胞色素C(Cytochrome C)、ERK1/2和p-ERK1/2(T202/204)蛋白表达。结果 2 m M、5 m M和10 m M浓度BHB对神经元细胞活力无显著影响(P>0.05),20 m M和50m M浓度BHB可致神经元活力显著降低(P<0.05)。神经元缺氧培养后,活力降低(P<0.05),线粒体氧化应激反应增强(P<0.05),SMCT1的m RNA水平和蛋白水平显著降低(P<0.05),caspase3与Cytochrome C的m RNA水平和蛋白水平明显增强(P<0.05),ERK1/2的蛋白磷酸化水平降低(P<0.05)。BHB预处理缺氧神经元,低浓度对细胞无明显影响(P>0.05);浓度增至10 m M,相比单纯缺氧组,神经元活力显著改善(P<0.05),线粒体氧化应激状态降低(P<0.05),SMCT1的m RNA水平和蛋白水平显著增加(P<0.05),caspase3与Cytochrome C的m RNA水平和蛋白水平明显降低(P<0.05),ERK1/2的蛋白磷酸化水平升高(P<0.05)。结论高浓度(>20 m M)BHB对神经元细胞有毒性作用;低浓度(<5 m M)BHB对糖氧剥夺的神经元细胞无明显作用;10 m M浓度的BHB预处理可增加SMCT1转运体的表达,通过启动ERK1/2信号通路,降低神经元线粒体的氧化应激,减少凋亡,从而对糖氧剥夺的原代神经元细胞发挥保护作用。展开更多
Monocarboxylic acid transporter 2 （MCT2） transports pyruvate and lactate outside and inside of sperms, mainly as energy sources and plays roles in the regulation of spermatogenesis. We investigated the association a...Monocarboxylic acid transporter 2 （MCT2） transports pyruvate and lactate outside and inside of sperms, mainly as energy sources and plays roles in the regulation of spermatogenesis. We investigated the association among genetic variations in the MCT2 gene, male infertility and MCT2 expression levels in sperm. The functional and genetic significance of the intron 2 （＋28201A 〉 G, rs10506398） and 3＇ untranslated region （UTR） single nucleotide polymorphism （SNP） （＋2626G 〉 A, rs10506399） of MCT2 variants were investigated. Two MCT2 polymorphisms were associated with male infertility （n = 471, P 〈 0.05）. In particular, the MCT2-3＇ UTR SNP （＋2626 G 〉 A） had a strong association with the oligoasthenoteratozoospermia （OAT） group. The ＋2626GG type had an almost 2.4-fold higher sperm count than that of the ＋2626AA type （＋2626GG; 66 x 106 vs ＋2626AA; 27 x 106, P 〈 0.0001）. The MCT2-3＇ UTR SNP may be important for expression, as it is located at the MCT2 3＇ UTR. The average MCT2 protein amount in sperm of the ＋2626GG type was about two times higher than that of the ＋2626AA type. The results suggest that genetic variation in MCT2 has functional and clinical relevance with male infertility.展开更多
文摘目的探讨β-羟基丁酸(BHB)对SD大鼠神经元细胞缺氧损伤的保护作用及其机制。方法原代培养SD大鼠神经元细胞,不同浓度(2 m M、5 m M、10 m M、20 m M和50 m M)BHB预处理24 h,三气培养箱糖氧剥夺培养2 h,线粒体超氧化物探针检测细胞线粒体氧化应激状态,CCK8检测细胞活力变化,QPCR测Na+偶联单羧酸转运体1(SMCT1)、caspae3和Cytochrome C的m RNA表达,Western Blot测SMCT1、caspase3、细胞色素C(Cytochrome C)、ERK1/2和p-ERK1/2(T202/204)蛋白表达。结果 2 m M、5 m M和10 m M浓度BHB对神经元细胞活力无显著影响(P>0.05),20 m M和50m M浓度BHB可致神经元活力显著降低(P<0.05)。神经元缺氧培养后,活力降低(P<0.05),线粒体氧化应激反应增强(P<0.05),SMCT1的m RNA水平和蛋白水平显著降低(P<0.05),caspase3与Cytochrome C的m RNA水平和蛋白水平明显增强(P<0.05),ERK1/2的蛋白磷酸化水平降低(P<0.05)。BHB预处理缺氧神经元,低浓度对细胞无明显影响(P>0.05);浓度增至10 m M,相比单纯缺氧组,神经元活力显著改善(P<0.05),线粒体氧化应激状态降低(P<0.05),SMCT1的m RNA水平和蛋白水平显著增加(P<0.05),caspase3与Cytochrome C的m RNA水平和蛋白水平明显降低(P<0.05),ERK1/2的蛋白磷酸化水平升高(P<0.05)。结论高浓度(>20 m M)BHB对神经元细胞有毒性作用;低浓度(<5 m M)BHB对糖氧剥夺的神经元细胞无明显作用;10 m M浓度的BHB预处理可增加SMCT1转运体的表达,通过启动ERK1/2信号通路,降低神经元线粒体的氧化应激,减少凋亡,从而对糖氧剥夺的原代神经元细胞发挥保护作用。
文摘Monocarboxylic acid transporter 2 （MCT2） transports pyruvate and lactate outside and inside of sperms, mainly as energy sources and plays roles in the regulation of spermatogenesis. We investigated the association among genetic variations in the MCT2 gene, male infertility and MCT2 expression levels in sperm. The functional and genetic significance of the intron 2 （＋28201A 〉 G, rs10506398） and 3＇ untranslated region （UTR） single nucleotide polymorphism （SNP） （＋2626G 〉 A, rs10506399） of MCT2 variants were investigated. Two MCT2 polymorphisms were associated with male infertility （n = 471, P 〈 0.05）. In particular, the MCT2-3＇ UTR SNP （＋2626 G 〉 A） had a strong association with the oligoasthenoteratozoospermia （OAT） group. The ＋2626GG type had an almost 2.4-fold higher sperm count than that of the ＋2626AA type （＋2626GG; 66 x 106 vs ＋2626AA; 27 x 106, P 〈 0.0001）. The MCT2-3＇ UTR SNP may be important for expression, as it is located at the MCT2 3＇ UTR. The average MCT2 protein amount in sperm of the ＋2626GG type was about two times higher than that of the ＋2626AA type. The results suggest that genetic variation in MCT2 has functional and clinical relevance with male infertility.
