目的:通过对C3H10T1/2细胞分化为成熟棕色脂肪细胞的培养、诱导分化及鉴定,深入探讨棕色脂肪细胞的生物学特性,为人类棕色脂肪细胞的相关研究提供实验参考与理论依据。方法:C3H10T1/2细胞经细胞接种、培养及诱导分化处理,利用光学显微...目的:通过对C3H10T1/2细胞分化为成熟棕色脂肪细胞的培养、诱导分化及鉴定,深入探讨棕色脂肪细胞的生物学特性,为人类棕色脂肪细胞的相关研究提供实验参考与理论依据。方法:C3H10T1/2细胞经细胞接种、培养及诱导分化处理,利用光学显微镜观察细胞形态变化,并通过油红O染色、免疫荧光法、线粒体探针法以及线粒体电镜技术对分化细胞进行鉴定分析。结果:未分化的C3H10T1/2细胞形态多样,具有突触伸展特征;分化后的细胞逐渐变为圆形或椭圆形,形成环形脂滴;未分化组细胞形态无明显变化。油红O染色显示,未分化组细胞基本无染色,而分化组中约90%的细胞红染,脂滴分布于细胞核周围,吸光度值显著升高(P<0.05)。分化组解偶联蛋白1(uncoupling protein 1,UCP1)、过氧化物酶体增殖激活受体γ(peroxisome proliferator-activated receptorγ,PPARγ)辅激活因子1α(PPARγco-activator-1α,PGC-1α)、PPARγ和转录因子PRDM16的相对荧光强度和蛋白表达量显著高于未分化组(P<0.05),且线粒体活性增强。结论:本研究成功诱导C3H10T1/2细胞分化为成熟棕色脂肪细胞,结合细胞形态观察、关键蛋白表达检测及线粒体功能分析进行综合评估,证实了细胞的有效分化及成熟棕色脂肪细胞的功能特性。展开更多
本文旨在探究富含天冬氨酸尾1的单通道膜蛋白(Single-pass Membrane Protein With Aspartate Rich Tail1,Smdt1)对C3H10T1/2细胞增殖和成脂分化的调控效应。本研究将Smdt1基因的过表达和干扰载体转染至C3H10T1/2细胞模型,采用qPCR方法...本文旨在探究富含天冬氨酸尾1的单通道膜蛋白(Single-pass Membrane Protein With Aspartate Rich Tail1,Smdt1)对C3H10T1/2细胞增殖和成脂分化的调控效应。本研究将Smdt1基因的过表达和干扰载体转染至C3H10T1/2细胞模型,采用qPCR方法量化了增殖和成脂分化关键基因的表达水平变化,利用EdU染色检测细胞增殖活力,油红O染色方法鉴定脂滴积累的状态;进一步通过String database、Bio GRID、Int Act、GeneMANIA、DAVID和Genecard数据库构建Smdt1蛋白互作网络图。结果显示,在C3H10T1/2细胞中过表达Smdt1,极显著提升了增殖标志基因Pcna、Ki67、Cdk1及Cdk4的表达,EdU阳性细胞比例反映了细胞增殖速率加快;Smdt1极显著促进成脂分化关键基因Pparγ、Fabp4、-Adipoq的表达量,显著促进了Cebpα、Cebpβ的表达量,脂滴数量变多。在C3H10T1/2细胞体系中,干扰Smdt1,与增殖紧密相关的标志基因,包括Ki67、Pcna及Cdk1,其表达水平极显著降低,Cdk4的表达也呈现显著降低的趋势,反映在EdU增殖检测中,阳性细胞数量明显减少,细胞增殖活性受到抑制。进一步干扰Smdt1后,成脂分化途径的关键调控基因Cebpα、Pparγ、Cebpβ、Fabp4、Adipoq的表达均极显著降低,细胞内脂滴的数量也显著减少,细胞成脂分化能力削弱。蛋白功能预测发现,Smdt1能够与Mcu相互作用,参与线粒体钙离子转运、摄取和稳态。本研究发现Smdt1可以促进C3H10T1/2细胞增殖和成脂分化,为脂肪沉积的研究提供了新的方向。展开更多
On January 30,2024,China announced the first human case of H10N5 influenza infection.Prior to this,human cases of H10N7 and H10N8 had been reported.It is now appropriate to re-examine the evolution and future epidemio...On January 30,2024,China announced the first human case of H10N5 influenza infection.Prior to this,human cases of H10N7 and H10N8 had been reported.It is now appropriate to re-examine the evolution and future epidemiological trends of the H10 and N5 subtypes of avian influenza viruses(AIVs).In this study,we analyzed the reassortment characteristics of the first human-derived H10N5 AIV(A/Zhejiang/ZJU01/2023),as well as the evolutionary dynamics of the wild bird-derived H10 and N5 subtypes of AIVs over the past decade.Our findings indicate that the human-derived H10N5 AIV exhibited low pathogenicity.A/bean_goose/Korea/KNU-10/2022(H10N7)and A/mallard/Novosibirsk_region/962k/2018(H12N5)were identified as the potential reassortment parents.The virus has existed since 2022 and several isolations have been reported in Bangladesh.Phylogenetic analysis showed that H10Ny and HxN5 AIVs in China are clustered differently based on the East Asian-Australian(eastern)and Central Asian-Indian(western)migratory flyways.The H10Ny and HxN5 AIV reassortant strains may cause human infections through accidental spillover.It is possible that another center of AIV evolution,mutation,and reassortment may be developing along the migratory flyways in northeastern Asia,distinct from Europe,the Americas,and China's Yangtze River Delta and Pearl River Delta,which should be closely monitored to ensure the safety of the public.展开更多
文摘目的:通过对C3H10T1/2细胞分化为成熟棕色脂肪细胞的培养、诱导分化及鉴定,深入探讨棕色脂肪细胞的生物学特性,为人类棕色脂肪细胞的相关研究提供实验参考与理论依据。方法:C3H10T1/2细胞经细胞接种、培养及诱导分化处理,利用光学显微镜观察细胞形态变化,并通过油红O染色、免疫荧光法、线粒体探针法以及线粒体电镜技术对分化细胞进行鉴定分析。结果:未分化的C3H10T1/2细胞形态多样,具有突触伸展特征;分化后的细胞逐渐变为圆形或椭圆形,形成环形脂滴;未分化组细胞形态无明显变化。油红O染色显示,未分化组细胞基本无染色,而分化组中约90%的细胞红染,脂滴分布于细胞核周围,吸光度值显著升高(P<0.05)。分化组解偶联蛋白1(uncoupling protein 1,UCP1)、过氧化物酶体增殖激活受体γ(peroxisome proliferator-activated receptorγ,PPARγ)辅激活因子1α(PPARγco-activator-1α,PGC-1α)、PPARγ和转录因子PRDM16的相对荧光强度和蛋白表达量显著高于未分化组(P<0.05),且线粒体活性增强。结论:本研究成功诱导C3H10T1/2细胞分化为成熟棕色脂肪细胞,结合细胞形态观察、关键蛋白表达检测及线粒体功能分析进行综合评估,证实了细胞的有效分化及成熟棕色脂肪细胞的功能特性。
文摘本文旨在探究富含天冬氨酸尾1的单通道膜蛋白(Single-pass Membrane Protein With Aspartate Rich Tail1,Smdt1)对C3H10T1/2细胞增殖和成脂分化的调控效应。本研究将Smdt1基因的过表达和干扰载体转染至C3H10T1/2细胞模型,采用qPCR方法量化了增殖和成脂分化关键基因的表达水平变化,利用EdU染色检测细胞增殖活力,油红O染色方法鉴定脂滴积累的状态;进一步通过String database、Bio GRID、Int Act、GeneMANIA、DAVID和Genecard数据库构建Smdt1蛋白互作网络图。结果显示,在C3H10T1/2细胞中过表达Smdt1,极显著提升了增殖标志基因Pcna、Ki67、Cdk1及Cdk4的表达,EdU阳性细胞比例反映了细胞增殖速率加快;Smdt1极显著促进成脂分化关键基因Pparγ、Fabp4、-Adipoq的表达量,显著促进了Cebpα、Cebpβ的表达量,脂滴数量变多。在C3H10T1/2细胞体系中,干扰Smdt1,与增殖紧密相关的标志基因,包括Ki67、Pcna及Cdk1,其表达水平极显著降低,Cdk4的表达也呈现显著降低的趋势,反映在EdU增殖检测中,阳性细胞数量明显减少,细胞增殖活性受到抑制。进一步干扰Smdt1后,成脂分化途径的关键调控基因Cebpα、Pparγ、Cebpβ、Fabp4、Adipoq的表达均极显著降低,细胞内脂滴的数量也显著减少,细胞成脂分化能力削弱。蛋白功能预测发现,Smdt1能够与Mcu相互作用,参与线粒体钙离子转运、摄取和稳态。本研究发现Smdt1可以促进C3H10T1/2细胞增殖和成脂分化,为脂肪沉积的研究提供了新的方向。
基金funded by the National Natural Science Foundation of China(81872673)the Shanghai New Three-year Action Plan for Public Health(GWVI-11.1-03).
文摘On January 30,2024,China announced the first human case of H10N5 influenza infection.Prior to this,human cases of H10N7 and H10N8 had been reported.It is now appropriate to re-examine the evolution and future epidemiological trends of the H10 and N5 subtypes of avian influenza viruses(AIVs).In this study,we analyzed the reassortment characteristics of the first human-derived H10N5 AIV(A/Zhejiang/ZJU01/2023),as well as the evolutionary dynamics of the wild bird-derived H10 and N5 subtypes of AIVs over the past decade.Our findings indicate that the human-derived H10N5 AIV exhibited low pathogenicity.A/bean_goose/Korea/KNU-10/2022(H10N7)and A/mallard/Novosibirsk_region/962k/2018(H12N5)were identified as the potential reassortment parents.The virus has existed since 2022 and several isolations have been reported in Bangladesh.Phylogenetic analysis showed that H10Ny and HxN5 AIVs in China are clustered differently based on the East Asian-Australian(eastern)and Central Asian-Indian(western)migratory flyways.The H10Ny and HxN5 AIV reassortant strains may cause human infections through accidental spillover.It is possible that another center of AIV evolution,mutation,and reassortment may be developing along the migratory flyways in northeastern Asia,distinct from Europe,the Americas,and China's Yangtze River Delta and Pearl River Delta,which should be closely monitored to ensure the safety of the public.
文摘2013年,从秦皇岛野鸟林鹬体内分离到一株H10N7亚型禽流感病毒(AIV),命名为A/Wood Sandpiper/Qinhuangdao/660-662/2013(H10N7)[简称WSP/QHD/660-662/2013(H10N7)]。本研究对该分离株的全基因序列进行测定,并对其进行致病性研究。基因组序列分析表明:该病毒的HA蛋白裂解位点为334PELMQGRGL343,属于低致病性AIV的分子特征,其HA基因与A/Duck/Hunan/S11205/2012(H10N3)的相似性达到97.90%,NA基因与A/Domestic Duck/Republic of Georgia/1/2010(H10N7)的相似性达到97.46%,内部基因与H9N2等多亚型AIV的相应基因节段具有较高的相似性,推测该分离株可能为一株多亚型流感病毒的重组株。对动物致病性试验结果显示:该病毒可以感染哺乳动物模型BALB/c小鼠,并且仅能够在鼠的肺脏和鼻甲骨粘膜上皮细胞中复制,表明该病毒分离株对小鼠也呈现低致病性。
