Lysophosphatidic acid(LPA)is a small molecule glycerophospholipid,which regulates multiple downstream signalling pathways through G-protein-coupled receptors to achieve numerous functions on oocyte maturation and embr...Lysophosphatidic acid(LPA)is a small molecule glycerophospholipid,which regulates multiple downstream signalling pathways through G-protein-coupled receptors to achieve numerous functions on oocyte maturation and embryo development.In this study,sheep in vitro fertilized embryos were applied to investigate the effects of LPA on early embryos development and embryonic stem cell establishment.At first,the maturation medium containing estrus female sheep serum and synthetic oviduct fluid(SOF)were optimized for sheep IVF,and then the effects of LPA were investigated.From 0.1 to 10μmol L^(–1),LPA had no significant effect on the cleavage rate(P>0.05),but the maturation rate and blastocyst rate increased dependently with LPA concentration(P<0.05),and the blastocyst morphology was normal.When the LPA concentration was 15μmol L^(–1),the maturation rate,cleavage rate and blastocyst rate decreased significantly(P<0.05),and the blastocyst exhibited abnormal morphology and could not develop into highquality blastocyst.Besides,the exogenous LPA increases the expression of LPAR2,LPAR4,TE-related gene CDX-2and pluripotency-related gene OCT-4 in sheep early IVF embryos with the raise of LPA concentration from 0.1 to 10μmol L^(–1).The expression of LPAR2,LPAR4,CDX-2 and OCT-4 from the LPA-0.1μmol L^(–1)to LPA-10μmol L^(–1)groups in early embryos were extremely significant(P<0.05),while the expression of these genes significantly decreased in 15μmol L^(–1)LPA-treated embryos compared with LPA-10μmol L^(–1)group(P<0.05).The inner cell mass in 15μmol L^(–1)LPA-treated embryos was also disturbed,and the blastocysts formation was abnormal.Secondly,the sheep IVF blastocysts were applied to establish embryonic stem cells.The results showed that LPA made the blastocyst inoculated cells grow towards TSC-like cells.They enhanced the fluorescence intensity and mRNA abundance of OCT-4 and CDX-2 as the concentration increased from 0 to 10μmol L^(–1),while 15μmol L^(–1)LPA decreased OCT-4 and CDX-2 expression in the derived cells.The expression of CDX-2 and OCT-4 in the blastocyst inoculated cells of LPA-1μmol L^(–1)group and LPA-10μmol L^(–1)group extremely significantly increased(P<0.05),but there was significant decrease in LPA-15μmol L^(–1)group compared with LPA-10μmol L^(–1)group(P<0.05).Meanwhile,the protein expression of LPAR2 and LPAR4 remarkably increased after treatment of LPA at 10μmol L^(–1)concentration.This study references the IVF embryo production and embryonic stem cell research of domestic animals.展开更多
目的探讨溶血磷脂酸(LPA)、Yes相关蛋白(YAP)及含PDZ结合区的共转录激活因子(TAZ)在头颈鳞癌中的表达及其与临床病理特征的关系。方法选择性收集雅安市人民医院病理科2010年1月至2016年12月期间,手术切除的病理组织石蜡标本共计136例,...目的探讨溶血磷脂酸(LPA)、Yes相关蛋白(YAP)及含PDZ结合区的共转录激活因子(TAZ)在头颈鳞癌中的表达及其与临床病理特征的关系。方法选择性收集雅安市人民医院病理科2010年1月至2016年12月期间,手术切除的病理组织石蜡标本共计136例,其中包括头颈鳞癌癌旁正常组织37例、头颈良性瘤样组织35例、头颈鳞癌64例,所有标本术前均无放化疗史,采用免疫组化及实时定量PCR检测LPA、YAP及TAZ在各组织中的表达及临床相关参数的关系。结果 LPA m RNA在头颈鳞癌癌旁正常组织、良性瘤样组织和鳞癌组织中的表达相近(P>0.05);YAP m RNA在鳞癌组织中的表达要高于癌旁组织、瘤样组织中的表达(P<0.05);TAZ m RNA在头颈3种组织中呈低表达,但鳞癌组织的表达明显高于良性瘤样组织及癌旁正常组织(P<0.05);YAP及TAZ蛋白在头颈鳞癌组织中的表达显著高于在癌旁正常组织和良性瘤样组织中的表达(P<0.05);鳞癌患者YAP的表达在肿瘤>3.0 cm、淋巴结阳性显著高于对照组(P<0.05);TAZ的表达在肿瘤中低分化、肿瘤>3.0 cm、淋巴结阳性及Ⅲ~Ⅳ期高于对照组(P<0.05)。结论 YAP及TAZ在头颈鳞癌中的表达与肿瘤分期、分化程度、淋巴结转移密切相关,而LPA的表达无显著差异及相关性。展开更多
基金financially supported by the Science and Technology Major Project of the Inner Mongolia Autonomous Region of China(2020ZD0007)the Major Program of the Inner Mongolia Natural Science Foundation,China(2020ZD10)+3 种基金the National Natural Science Foundation of China(32160172)the Natural Science Foundation of Inner Mongolia Autonomous Region(2020BS03003 and 2020BS03022)the National Transgenic Project of China(2016ZX0801000-002 and 2016ZX08010005-001)the Science and Technology Major Project of the Inner Mongolia Autonomous Region of China(zdzx2018065)。
文摘Lysophosphatidic acid(LPA)is a small molecule glycerophospholipid,which regulates multiple downstream signalling pathways through G-protein-coupled receptors to achieve numerous functions on oocyte maturation and embryo development.In this study,sheep in vitro fertilized embryos were applied to investigate the effects of LPA on early embryos development and embryonic stem cell establishment.At first,the maturation medium containing estrus female sheep serum and synthetic oviduct fluid(SOF)were optimized for sheep IVF,and then the effects of LPA were investigated.From 0.1 to 10μmol L^(–1),LPA had no significant effect on the cleavage rate(P>0.05),but the maturation rate and blastocyst rate increased dependently with LPA concentration(P<0.05),and the blastocyst morphology was normal.When the LPA concentration was 15μmol L^(–1),the maturation rate,cleavage rate and blastocyst rate decreased significantly(P<0.05),and the blastocyst exhibited abnormal morphology and could not develop into highquality blastocyst.Besides,the exogenous LPA increases the expression of LPAR2,LPAR4,TE-related gene CDX-2and pluripotency-related gene OCT-4 in sheep early IVF embryos with the raise of LPA concentration from 0.1 to 10μmol L^(–1).The expression of LPAR2,LPAR4,CDX-2 and OCT-4 from the LPA-0.1μmol L^(–1)to LPA-10μmol L^(–1)groups in early embryos were extremely significant(P<0.05),while the expression of these genes significantly decreased in 15μmol L^(–1)LPA-treated embryos compared with LPA-10μmol L^(–1)group(P<0.05).The inner cell mass in 15μmol L^(–1)LPA-treated embryos was also disturbed,and the blastocysts formation was abnormal.Secondly,the sheep IVF blastocysts were applied to establish embryonic stem cells.The results showed that LPA made the blastocyst inoculated cells grow towards TSC-like cells.They enhanced the fluorescence intensity and mRNA abundance of OCT-4 and CDX-2 as the concentration increased from 0 to 10μmol L^(–1),while 15μmol L^(–1)LPA decreased OCT-4 and CDX-2 expression in the derived cells.The expression of CDX-2 and OCT-4 in the blastocyst inoculated cells of LPA-1μmol L^(–1)group and LPA-10μmol L^(–1)group extremely significantly increased(P<0.05),but there was significant decrease in LPA-15μmol L^(–1)group compared with LPA-10μmol L^(–1)group(P<0.05).Meanwhile,the protein expression of LPAR2 and LPAR4 remarkably increased after treatment of LPA at 10μmol L^(–1)concentration.This study references the IVF embryo production and embryonic stem cell research of domestic animals.
文摘目的探讨溶血磷脂酸(LPA)、Yes相关蛋白(YAP)及含PDZ结合区的共转录激活因子(TAZ)在头颈鳞癌中的表达及其与临床病理特征的关系。方法选择性收集雅安市人民医院病理科2010年1月至2016年12月期间,手术切除的病理组织石蜡标本共计136例,其中包括头颈鳞癌癌旁正常组织37例、头颈良性瘤样组织35例、头颈鳞癌64例,所有标本术前均无放化疗史,采用免疫组化及实时定量PCR检测LPA、YAP及TAZ在各组织中的表达及临床相关参数的关系。结果 LPA m RNA在头颈鳞癌癌旁正常组织、良性瘤样组织和鳞癌组织中的表达相近(P>0.05);YAP m RNA在鳞癌组织中的表达要高于癌旁组织、瘤样组织中的表达(P<0.05);TAZ m RNA在头颈3种组织中呈低表达,但鳞癌组织的表达明显高于良性瘤样组织及癌旁正常组织(P<0.05);YAP及TAZ蛋白在头颈鳞癌组织中的表达显著高于在癌旁正常组织和良性瘤样组织中的表达(P<0.05);鳞癌患者YAP的表达在肿瘤>3.0 cm、淋巴结阳性显著高于对照组(P<0.05);TAZ的表达在肿瘤中低分化、肿瘤>3.0 cm、淋巴结阳性及Ⅲ~Ⅳ期高于对照组(P<0.05)。结论 YAP及TAZ在头颈鳞癌中的表达与肿瘤分期、分化程度、淋巴结转移密切相关,而LPA的表达无显著差异及相关性。
