为揭示幽门螺杆菌(Helicobacter pylori)感染胃黏膜上皮细胞来源的外泌体(exosome)对巨噬细胞极化的影响,分析其蛋白质的组分和功能,收集感染与未感染幽门螺杆菌的胃黏膜上皮细胞GES-1上清液中的外泌体;利用蛋白质印迹法、流式细胞术观...为揭示幽门螺杆菌(Helicobacter pylori)感染胃黏膜上皮细胞来源的外泌体(exosome)对巨噬细胞极化的影响,分析其蛋白质的组分和功能,收集感染与未感染幽门螺杆菌的胃黏膜上皮细胞GES-1上清液中的外泌体;利用蛋白质印迹法、流式细胞术观察外泌体对巨噬细胞极化的影响;通过蛋白质组学方法分析鉴定出感染与未感染幽门螺杆菌的胃黏膜上皮细胞GES-1来源的外泌体中的差异蛋白质,并采用蛋白质印迹法验证正常胃黏膜上皮细胞(GES-1)和胃癌细胞(MKN-7、AGS)中差异蛋白质的表达;最后,运用免疫组织化学法分析两组临床组织标本的热休克蛋白家族D成员1(heat shock protein family D member 1,HSPD1)水平,以及M2型巨噬细胞标志物CD163的表达水平。结果显示,幽门螺杆菌感染胃黏膜上皮细胞所释放的外泌体促进M2型巨噬细胞的表面标志物CD163、CD206以及极化相关蛋白质MGL1/2和Arg-1的表达;蛋白质组学共筛选出50个差异表达的蛋白质,其中HSPD1在幽门螺杆菌感染后的GES-1细胞及其释放的外泌体中的表达水平显著高于未感染组。此外,幽门螺杆菌阳性胃癌组织的HSPD1表达水平显著高于幽门螺杆菌阴性胃癌组织;在幽门螺杆菌阳性胃癌组织中,M2型巨噬细胞标志物CD163的表达水平显著增加,且与HSPD1蛋白的表达水平呈明显正相关(r=0.472,P=0.01)。这些研究结果表明,幽门螺杆菌感染胃黏膜上皮细胞来源的外泌体促进巨噬细胞M2型极化。其中,外泌体携带的HSPD1作为关键调控因子,在驱动巨噬细胞表型转化进程中发挥着不可或缺的作用。展开更多
Background:Sperm quality evaluation is the logical first step in increasing field fertility.Spermatozoa contain cytoplasmic organelles and biomolecules known as sperm-intrinsic factors,which play key roles in sperm ma...Background:Sperm quality evaluation is the logical first step in increasing field fertility.Spermatozoa contain cytoplasmic organelles and biomolecules known as sperm-intrinsic factors,which play key roles in sperm maturation,sperm-oocyte fusion,and embryo development.In particular,sperm membrane proteins[e.g.,arginine vasopressin receptor 2,beta-actin,prohibitin,and heat shock protein family D member 1(HSPD1)]and RNA could be used as functional indicators of male fertility.We sought to clarify the effects of differential mRNA expression of selected genes on several fertilisation parameters,including sperm motility,motion kinematics,capacitation,and litter size,in a porcine model.Results:Our results demonstrated that HSPD1 expression was significantly correlated with male fertility,as measured by the litter size of inseminated sows.The expression of HSPD1 mRNA was linked to sperm motility and other motion kinematic characteristics.Furthermore,HSPD1 had a 66.7%overall accuracy in detecting male fertility,and the high-litter size group which was selected with the HSPD1 marker had a 1.34 greater litter size than the lowlitter size group.Conclusions:Our findings indicate that HSPD1 might be a helpful biomarker for superior boar selection for artificial insemination,which could boost field fertility.展开更多
文摘为揭示幽门螺杆菌(Helicobacter pylori)感染胃黏膜上皮细胞来源的外泌体(exosome)对巨噬细胞极化的影响,分析其蛋白质的组分和功能,收集感染与未感染幽门螺杆菌的胃黏膜上皮细胞GES-1上清液中的外泌体;利用蛋白质印迹法、流式细胞术观察外泌体对巨噬细胞极化的影响;通过蛋白质组学方法分析鉴定出感染与未感染幽门螺杆菌的胃黏膜上皮细胞GES-1来源的外泌体中的差异蛋白质,并采用蛋白质印迹法验证正常胃黏膜上皮细胞(GES-1)和胃癌细胞(MKN-7、AGS)中差异蛋白质的表达;最后,运用免疫组织化学法分析两组临床组织标本的热休克蛋白家族D成员1(heat shock protein family D member 1,HSPD1)水平,以及M2型巨噬细胞标志物CD163的表达水平。结果显示,幽门螺杆菌感染胃黏膜上皮细胞所释放的外泌体促进M2型巨噬细胞的表面标志物CD163、CD206以及极化相关蛋白质MGL1/2和Arg-1的表达;蛋白质组学共筛选出50个差异表达的蛋白质,其中HSPD1在幽门螺杆菌感染后的GES-1细胞及其释放的外泌体中的表达水平显著高于未感染组。此外,幽门螺杆菌阳性胃癌组织的HSPD1表达水平显著高于幽门螺杆菌阴性胃癌组织;在幽门螺杆菌阳性胃癌组织中,M2型巨噬细胞标志物CD163的表达水平显著增加,且与HSPD1蛋白的表达水平呈明显正相关(r=0.472,P=0.01)。这些研究结果表明,幽门螺杆菌感染胃黏膜上皮细胞来源的外泌体促进巨噬细胞M2型极化。其中,外泌体携带的HSPD1作为关键调控因子,在驱动巨噬细胞表型转化进程中发挥着不可或缺的作用。
基金supported by the Basic Science Research Program through the National Research Foundation of Korea(NRF),funded by the Ministry of Education(NRF-2018R1A6A1A03025159).
文摘Background:Sperm quality evaluation is the logical first step in increasing field fertility.Spermatozoa contain cytoplasmic organelles and biomolecules known as sperm-intrinsic factors,which play key roles in sperm maturation,sperm-oocyte fusion,and embryo development.In particular,sperm membrane proteins[e.g.,arginine vasopressin receptor 2,beta-actin,prohibitin,and heat shock protein family D member 1(HSPD1)]and RNA could be used as functional indicators of male fertility.We sought to clarify the effects of differential mRNA expression of selected genes on several fertilisation parameters,including sperm motility,motion kinematics,capacitation,and litter size,in a porcine model.Results:Our results demonstrated that HSPD1 expression was significantly correlated with male fertility,as measured by the litter size of inseminated sows.The expression of HSPD1 mRNA was linked to sperm motility and other motion kinematic characteristics.Furthermore,HSPD1 had a 66.7%overall accuracy in detecting male fertility,and the high-litter size group which was selected with the HSPD1 marker had a 1.34 greater litter size than the lowlitter size group.Conclusions:Our findings indicate that HSPD1 might be a helpful biomarker for superior boar selection for artificial insemination,which could boost field fertility.
