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Regulatory Role of Retinoic Acid in Male Pregnancy of the seahorse

The Innovation 2020年第3期1卷 72-80页

作者： Chunyan Li Yongxin Li Geng Qin Zelin Chen Meng Qu Bo Zhang Xue Han Xin Wang Pei-yuan Qian Qiang Lin CAS Key Laboratory of Tropical Marine Bio-Resources and Ecology South China Sea Institute of OceanologyInnovation of South China Sea Ecology and Environmental EngineeringChinese Academy of Sciences510301 GuangzhouChina Southern Marine Science and Engineering Guangdong Laboratory(Guangzhou) 511458 GuangzhouChina Department of Chemistry The University of Hong KongPokfulamHong KongHong Kong SARChina Department of Ocean Science and Hong Kong Branch of Southern Marine Science and Engineering Guangdong Laboratory(Guangzhou) Hong Kong University of Science and TechnologyKowloonHong KongChina

seahorses epitomize the exuberance of *** have the unique characteristic of male pregnancy,which includes the carrying of many embryos in a brood pouch that incubates and nourishes the embryos,similar to the mammalian ***,the regulatory networks underlying brood pouch formation and pregnancy remain largely *** this study,comparative transcriptomic and metabolomic profiling on the lined seahorse Hippocampus erectus,with unformed,newly formed,and pregnant brood pouches identified a total of 141 and 2,533 differentially expressed genes together with 73 and 121 significantly differential metabolites related to brood pouch formation and pregnancy,***,integrative omics analysis revealed that retinoic acid(RA)synthesis and signaling pathway played essential roles in the formation of the brood pouch and *** might function upstream of testosterone and progesterone,thereby directly influencing brood pouch formation by regulating the expression of fshr and *** results also revealed that RA regulates antioxidant defenses,particularly during male ***,pregnancy caused a consistent decrease in RA,canthaxanthin,astaxanthin,and glutathione synthetase,and an increase in susceptibility to oxidative stress,which may balance brood pouch development and reproduction in seahorses and pave the way to successful gestation.

关键词： seahorse MALE PREGNANCY BROOD POUCH INTEGRATIVE OMICS RETINOIC ACID ANTIOXIDANT DEFENSE

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Wnt8a is one of the candidate genes that play essential roles in the elongation of the seahorse prehensile tail

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Marine Life Science & Technology 2021年第4期3卷 416-426页

作者： Bo Zhang Geng Qin Lili Qu Yanhong Zhang Chunyan Li Chunlei Cang Qiang Lin CAS Key Laboratory of Tropical Marine Bio-Resources and Ecology South China Sea Institute of OceanologyChinese Academy of SciencesGuangzhou 510275China Institution of South China Sea Ecology and Environmental Engineering Chinese Academy of SciencesGuangzhou 510275China Southern Marine Science and Engineering Guangdong Laboratory(Guangzhou) Guangzhou 511458China School of Life Sciences University of Science and Technology of ChinaHefei 230026China University of Chinese Academy of Sciences Beijing 100049China

seahorses are a hallmark of specialized morphological features due to their elongated prehensile ***,the underlying genomic grounds of seahorse tail development remain ***,we evaluated the roles of essential genes from the Wnt gene family for the tail developmental process in the lined seahorse(Hippocampus erectus).Comparative genomic analysis revealed that the Wnt gene family is conserved in *** expression profiles and in situ hybridization suggested that Wnt5a,Wnt8a,and Wnt11 may participate in seahorse tail *** in other teleosts,Wnt5a and Wntll were found to regulate the development of the tail axial mesoderm and tail somitic mesoderm,***,a significantly extended expression period of Wnt8a during seahorse tail development was *** pathway analysis further showed that Wnt5a up-regulated the expression of the tail axial mesoderm gene(Shh),while interaction analysis indicated that Wnt8a could promote the expression of *** summary,our results indicate that the special extended expression period of Wnt8a might promote caudal tail axis formation,which contributes to the formation of the elongated tail of the seahorse.

关键词： Embryo seahorse Tail development Wnt8a Wnt gene family

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利用seahorse XF-96探讨不同细胞浓度和血清对线粒体功能的影响

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局解手术学杂志 2022年第4期31卷 284-291页

作者：李晓栩崔宇阳一栋周晓英矫力王辰元杨诚忠史诗黄缄陆军军医大学高原军事医学系/高原生理学与病理学教研室/极端环境医学教育部重点实验室/全军高原医学重点实验室重庆400038 重庆医药高等专科学校生理教研室重庆401331

目的利用seahorse XF-96探讨不同细胞浓度和血清对HUVEC细胞和U87细胞线粒体功能的影响。方法利用细胞线粒体应激试剂盒,在不同浓度的HUVEC细胞和U87细胞中加入抑制剂(寡霉素、解偶联剂、鱼藤酮和抗霉素A),根据细胞浓度分为2000组、400... 详细信息

目的利用seahorse XF-96探讨不同细胞浓度和血清对HUVEC细胞和U87细胞线粒体功能的影响。方法利用细胞线粒体应激试剂盒,在不同浓度的HUVEC细胞和U87细胞中加入抑制剂(寡霉素、解偶联剂、鱼藤酮和抗霉素A),根据细胞浓度分为2000组、4000组、6000组和8000组,根据有无血清和不同的解偶联剂浓度分为血清+低解偶联剂组、血清+高解偶联剂组、无血清+低解偶联剂组和无血清+高解偶联剂组。在seahorse XF-96中测定线粒体的耗氧率(OCR)和细胞外酸化率(ECAR),并比较HUVEC细胞和U87细胞各组线粒体的基础氧耗、质子漏、最大呼吸能力、非线粒体的氧耗、储备的呼吸能力、三磷酸腺苷(ATP)生成能力、线粒体呼吸潜力和偶联效率;比较HUVEC细胞和U87细胞之间上述指标的差异;观察血清和解偶联剂浓度对HUVEC细胞线粒体功能的影响。结果在HUVEC细胞和U87细胞中,2000组OCR显著低于4000组、6000组和8000组(P0.05)。U87细胞8000组的质子漏高于HUVEC细胞(P0.05)。血清+低解偶联剂组储备的呼吸能力和线粒体呼吸潜力高于无血清+低解偶联剂组(P0.05);血清+高解偶联剂组和无血清+高解偶联剂组的线粒体功能各指标差异均无统计学意义(P>0.05)。结论随着细胞浓度的不断增加,HUVEC细胞和U87细胞的OCR和ECAR不断升高,细胞浓度会影响两种细胞线粒体的OCR和ECAR,HUVEC细胞和U87细胞的质子漏和偶联效率存在显著差异,血清会影响HUVEC细胞线粒体储备的呼吸能力和线粒体呼吸潜力。

关键词：线粒体能量代谢氧耗 seahorse

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An examination of the population dynamics of syngnathid fishes within Tampa Bay,Florida,USA

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Current Zoology 2010年第1期56卷 118-133页

作者： Heather D.MASONJONES Emily ROSE Lori Benson McRAE Danielle L.DIXSON Biology Department University of Tampa Biology Department Texas A & M University School of Marine and Tropical Biology James Cook University

Seagrass ecosystems worldwide have been declining, leading to a decrease in associated fish populations, especially those with low mobility such as syngnathids （pipefish and seahorses）. This two-year pilot study investigated seasonal patterns in density, growth, site fidelity, and population dynamics of Tampa Bay （FL） syngnathid fishes at a site adjacent to two marinas under construction. Using a modified mark-recapture technique, fish were collected periodically from three closely located sites that varied in seagrass species （Thalassia spp., Syringodium spp., and mixed-grass sites） and their distance from open water, but had consistent physical/chemical environmental characteristics. Fish were marked, photographed for body size and gender measurements, and released the same day at the capture site. Of the 5695 individuals surveyed, 49 individuals were recaptured, indicating a large, flexible population. Population density peaks were observed in July of both years, with low densities in late winter and late summer. Spatially, syngnathid densities were highest closest to the mouth of the bay and lowest near the shoreline. Seven species of syngnathid fishes were observed, and species-specific patterns of seagrass use emerged during the study. However, only two species, Syngnathus scovelli and Hippoeampus zosterae, were observed at high frequencies. For these two species, body size decreased across the study period, but while S. scovelli＇s population density decreased, H. zosterae＇s increased. Across six of the seven species, population size declined over the course of this preliminary study; however, seasonal shifts were impossible to distinguish from potential anthropogenic effects of construction [Current Zoology 56 （1）： 118-133, 2010].

关键词： Syngnathids Habitat-use seahorse Pipefish Population density Mark-recapture

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Evolutionary traits and functional roles of chemokines and their receptors in the male pregnancy of the Syngnathidae

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Marine Life Science & Technology 2023年第4期5卷 500-510页

作者： Han Jiang Zhanwei Zhao Haiyan Yu Qiang Lin Yali Liu CAS Key Laboratory of Tropical Marine Bio-Resources and Ecology South China Sea Institute of OceanologyChinese Academy of SciencesGuangzhou 510301China Guangdong Provincial Key Laboratory of Applied Marine Biology South China Sea Institute of OceanologyChinese Academy of SciencesGuangzhou 510301China University of Chinese Academy of Sciences Beijing 101400China

Vertebrates have developed various modes of reproduction,some of which are found in *** 300 species of the Syngnathidae(seahorses,pipefishes and seadragons)exhibit male pregnancies;the males have specialized brood pouches that provide immune protection,nourishment,and oxygen *** play a vital role at the mammalian maternal–fetal interface;however,their functions in fish reproduction are *** study revealed the evolutionary traits and potential functions of chemokine genes in 22 oviparous,ovoviviparous,and viviparous fish species through comparative genomic *** results showed that chemokine gene copy numbers and evolutionary rates vary among species with different modes of *** lost cxcl13 and cxcr5,which are involved in key receptor–ligand pairs for lymphoid organ ***,Syngnathidae have site-specific mutations in cxcl12b and ccl44,suggesting immune function during ***,transcriptome analysis revealed that chemokine gene expression varies among Syngnathidae species with different types of brood pouches,suggesting adaptive variations in chemokine functions among seahorses and their ***,challenge experiments on seahorse brood pouches revealed a joint immune function of chemokine genes during male *** study provides insights into the evolutionary diversity of chemokine genes associated with different reproductive modes in fish.

关键词： Reproductive mode Immune response seahorse Brood pouch

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Pharmacological efficacy of the traditional Chinese medicinal formula Kun-Tai-1A in the treatment of letrozole-induced polycystic ovary syndrome

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Traditional Medicine Research 2022年第3期7卷 44-54页

作者： Leanne Lee Leung Jing Xie Ya-Cun Chen Kar-Ho Lam Hei Wan Su-Lan Yu Tzi-Bun Ng George Pak-Heng Leung Jin Yu Ren-Min Yao Shu-Jia Sun Sydney Chi-Wai Tang Hai-Yong Chen Jia Zhao Zhang-Jin Zhang Calvin Kai-Fai Lee Kalin Yan-Bo Zhang Li-Xing Lao Yun Feng Xiang Lin Wei Meng School of Chinese Medicine Li Ka Shing Faculty of MedicineThe University of Hong KongHong Kong 999077China School of Biomedical Sciences Faculty of MedicineThe Chinese University of Hong KongHong Kong 999077China Department of Pharmacology and Pharmacy Li Ka Shing Faculty of MedicineThe University of Hong KongHong Kong 999077China Workstation for Training and Research(Hong Kong Branch) Distinguished Professor Yu Jin Gynaecology of Chinese Medicine&Integrative MedicineHong Kong 999077China Shanghai Taikuntang Chinese Medicine Hospital Shanghai 200023China Department of Medicine Li Ka Shing Faculty of MedicineThe University of Hong KongHong Kong 999077China Department of Obstetrics and Gynaecology Li Ka Shing Faculty of MedicineThe University of Hong KongHong Kong 999077China Virginia University of Integrative Medicine Virginia 22031USA Department of Ophthalmology Peking University Third HospitalBeijing 100191China Workstation of Zhu Nansun National Master of Chinese MedicineHong Kong Branch of Zhu’s School of Gynaecology of Chinese Medicine from ShanghaiHong Kong 999077China

Background:Polycystic ovary syndrome(PCOS)is an endocrine disorder that occurs in women of child-bearing ***,PCOS patients have decreased pregnancy rates and clomiphene citrate *** traditional Chinese medicine formula Kun-Tai-1S(KT1S),consisting of the seahorse species hippocampus,has been reported to elicit therapeutic effects in patients with ***,given the limited resources and global demand for wild hippocampus,whether KT1S with or without hippocampus can elicit similar therapeutic effects has not been ***:KT1S and Kun-Tai-1A(KT1A,KT1S without dry hippocampus)were used to treat a letrozole-induced rat model of PCOS with an established *** serum levels of testosterone,luteinizing hormone,anti-Müllerian hormone,and estradiol were determined,the luteinizing hormone/follicle-stimulating hormone ratio was determined,and the ovarian pathology was ***:Similar to the therapeutic effects of cyproterone acetate,both the KT1S and KT1A treatments reduced the body weight and ovarian and uterine indices in the rats with *** serum levels of testosterone,anti-Müllerian hormone,and luteinizing hormone and the luteinizing hormone/follicle-stimulating hormone ratio were significantly lower in the KT1S and KT1A treatment groups compared to the model group(P<0.01 and P<0.05,respectively).Moreover,the histopathological assessment results suggested that both the KT1S and KT1A treatments significantly ameliorated the PCOS pathology in the rats with an established disease,with a reduced number of cystic and atretic follicles and an increased number of corpora lutea being observed in the ***,there was no obvious difference in the disease outcomes between the KT1S and KT1A-treated *** pharmacology analysis revealed that 4’,7-dihydroxyflavanone,sinpemine A,quercetin,8-isopentenyl-kaempferol,and luteolin in KT1A may promote estrogen signaling;furthermore,the nitric oxide regulation pathway is also cl

关键词： polycystic ovary syndrome traditional Chinese medicine ovulation Kun-Tai-1A seahorse endocrine disease

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基于能量代谢分析益气解毒方对缺氧损伤的PC12细胞线粒体保护作用的机制

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中国实验方剂学杂志 2022年第4期28卷 34-41页

作者：戴玉豪刘黎明刘陈吴文洁沈建英李韶菁中国中医科学院中药研究所北京100700 安徽中医药大学合肥230000

目的:采用seahorse细胞能量代谢分析仪,通过缺氧损伤的高分化大鼠肾上腺嗜铬瘤细胞(PC12细胞)建立一种线粒体能量代谢评价方法并探究益气解毒方的线粒体保护作用。方法:应用缺氧小室体外构建PC12细胞损伤模型,实验设置空白组,模型组(Mod... 详细信息

目的:采用seahorse细胞能量代谢分析仪,通过缺氧损伤的高分化大鼠肾上腺嗜铬瘤细胞(PC12细胞)建立一种线粒体能量代谢评价方法并探究益气解毒方的线粒体保护作用。方法:应用缺氧小室体外构建PC12细胞损伤模型,实验设置空白组,模型组(Model),益气解毒方高、中、低剂量组(25,5,1μmol·L^(-1);YQ高、中、低)及阳性药曲美他嗪组(TMZ),每组3复孔,实验重复3次。采用建立的能量代谢分析方法对细胞线粒体复合物活性进行检测,筛选最佳给药浓度;随后设置益气解毒方全方及各拆方组,采用seahorse细胞能量代谢分仪检测有氧呼吸和糖酵解功能;根据非线粒体呼吸值,质子漏值,基础呼吸值,最大呼吸值,ATP生成量,可提升呼吸能力检测值,通过主成份分析(PCA)和变量重要性指标(VIP),分析评价益气解毒方不同配伍组对缺氧损伤线粒体有氧呼吸作用效果;通过蛋白免疫印迹法(Western blot)检测细胞色素C(Cytochrome C),B细胞淋巴瘤-2(Bcl-2)和Bcl-2相关X蛋白(Bax)的表达。结果:与其他浓度组比较,FCCP的最佳给药浓度为2μmol·L^(-1);与模型组比较,5μmol·L^(-1)益气解毒方全方能提升线粒体复合物活性(P<0.05);与模型组比较,益气解毒方全方改善效果较好(P<0.01);在各拆方组中,人参皂苷和栀子苷配伍效果较好(P<0.01);VIP分析结果显示,对于线粒体呼吸功能改善,益气解毒方中栀子苷贡献度最大;与模型组比较,益气解毒方中、高剂量组线粒体细胞色素C的渗漏显著减少(P<0.01),Bax蛋白的表达显著降低(P<0.01),Bcl-2蛋白的表达明显升高(P<0.05,P<0.01)。结论:建立了一种细胞水平、高通量线粒体能量代谢多指标定量评价方法,证明了益气解毒方全方能够显著改善缺氧损伤后PC12细胞线粒体能量代谢障碍,而发挥这一作用的机制可能与线粒体凋亡保护相关。

关键词：益气解毒方配伍 seahorse 线粒体能量代谢缺氧模型

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缺氧对小鼠骨髓间充质干细胞体外培养及成骨分化影响的研究

缺氧对小鼠骨髓间充质干细胞体外培养及成骨分化影响的研究

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作者：王许巧兰州大学

学位级别：硕士

目的:观察1%O缺氧环境对小鼠骨髓间充质干细胞(Bone mesenchymal stem cells,BMSCs)体外培养、增殖、迁移及成骨分化的影响并探讨相关分子机制。方法:(1)小鼠BMSCs的体外培养:选择全骨髓贴壁法分离小鼠原代细胞,分别于1%O和21%O条件下... 详细信息

目的:观察1%O缺氧环境对小鼠骨髓间充质干细胞(Bone mesenchymal stem cells,BMSCs)体外培养、增殖、迁移及成骨分化的影响并探讨相关分子机制。方法:(1)小鼠BMSCs的体外培养:选择全骨髓贴壁法分离小鼠原代细胞,分别于1%O和21%O条件下纯化、培养细胞,在显微镜下观察、记录细胞的形态和增殖情况;(2)1%O和21%O条件下小鼠BMSCs分别向成骨样、成脂样和成神经样细胞方向诱导分化:将P3代的小鼠BMSCs用成骨样细胞诱导液、成脂样细胞诱导液以及成神经样细胞诱导液分别诱导培养14,21和3天,在显微镜下观察记录细胞形态和增殖分化状况,然后将成骨样细胞用茜素红染液染色、成脂样细胞用油红O染液染色、成神经样用细胞尼氏染色液染色鉴定;(3)流式细胞术鉴定1%O和21%O组小鼠BMSCs:选取P3代的1%O和21%O组小鼠BMSCs,待细胞长至80%-90%融合度时,制备成单细胞悬液,上机检测Sca-1、CD29、CD117、CD45表面分子标记物;(4)1%O和21%O组小鼠BMSCs的检测指标:CCK-8法检测各组小鼠BMSCs 1到7天的增殖情况,制作增殖曲线;细胞划痕实验检测1%O和21%O组小鼠BMSCs在0h,6h,12h,24h和48h的细胞迁移效率;Mito Tracker染色后荧光显微镜观察不同培养条件下线粒体结构和形态的变化,并用流式细胞术检测荧光强度;将1%O和21%O组细胞均用成骨诱导液连续诱导3,5,7,10和14天,检测上清液中碱性磷酸酶(Alkaline phosphatase,ALP)的活性,并在成骨诱导液诱导14天后茜素红染色,在显微镜下观察成骨效率,同时,用氯化十六烷基吡啶溶解钙结节进行半定量检测;seahorse细胞代谢能量分析仪检测各组细胞耗氧率(Oxygen consumption rate,OCR);Western blot检测成骨相关基因Run相关转录因子2(Runt-related transcription factor 2,Runx2)和骨桥蛋白的(Osteopontin,OPN)表达。结果:(1)选用全骨髓贴壁法获得的小鼠BMSCs,在1%O和21%O条件下培养均为纺锤形、多角形克隆样生长的细胞;(2)小鼠BMSCs经过成骨诱导液、成脂诱导液以及成神经诱导液的条件分化后,三组染色结果均为阳性,分别观察到明显的红色矿化结节、红色脂滴和深紫色尼氏小体;(3)流式细胞分析鉴定结果显示在1%O和21%O条件下培养获得的细胞,Sca-1、CD29表达均为阳性,而CD117、CD45表达均为阴性;(4)细胞增殖实验结果显示,在1%O和21%O条件下培养获得的细胞,其增殖效率第1、2天时两组之间无显著性差异,第3天至第7天时1%O组的细胞增殖效率明显高于21%O组;细胞划痕实验表明,与21%O组相比,1%O可明显促进细胞迁移效率;Mito Tracker染色后发现1%O组线粒体结构形态较21%O组成熟,呈短棒状,连接成网状,流式结果表明1%O组Mito Tracker染色荧光强度明显强于21%O组;茜素红染色发现1%O组钙结节明显多于21%O组;碱性磷酸酶检测表明第7天1%O组ALP活性表达明显高于21%O组;seahorse检测结果表明1%O组OCR高于21%O组;Western blot结果表明1%组BMSCs的成骨相关蛋白RUNX2和OPN表达明显高于21%O组,且以上结果均具有统计学差异(p<0.05)。结论:1%O和21%O条件下获得的小鼠原代细胞均为具有干细胞特征的骨髓间充质干细胞,然而,缺氧条件下获得的小鼠BMSCs具有更高的增殖效率和迁移效率,同时缺氧条件下培养的小鼠BMSCs具有更高的成骨分化效率,且分化效率与细胞线粒体结构形态及线粒体耗氧率结果保持一致。

关键词：小鼠骨髓间充质干细胞 1%O2 成骨分化线粒体 seahorse

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细胞能量代谢分析法评价对乙酰氨基酚对人肝细胞线粒体有氧呼吸的毒性作用

引用

中国药物警戒 2019年第4期16卷 193-198,207页

作者：李梅刘宁张凡孙华中国医学科学院北京协和医学院药物研究所北京100050

目的利用seahorse XF^e24海马细胞能量代谢分析系统分析对乙酰氨基酚(APAP)对人肝细胞线粒体有氧呼吸的整体影响,为APAP在能量代谢角度的毒性防控及毒性机制研究提供方法学的参考。方法体外培养人肝细胞系(HepG2),分别以1、2、4、8、16 ... 详细信息

目的利用seahorse XF^e24海马细胞能量代谢分析系统分析对乙酰氨基酚(APAP)对人肝细胞线粒体有氧呼吸的整体影响,为APAP在能量代谢角度的毒性防控及毒性机制研究提供方法学的参考。方法体外培养人肝细胞系(HepG2),分别以1、2、4、8、16 mM APAP处理细胞24 h,MTT法检测细胞增殖情况,倒置显微镜观察细胞形态;BCA法定蛋白含量。采用seahorse XF^e24海马细胞能量代谢分析系统建立HepG2细胞实验方法,并检测不同浓度APAP对人肝细胞线粒体有氧呼吸的影响和特点。结果 seahorse XF^e24海马细胞能量代谢分析系统检测HepG2细胞能量代谢的最佳条件为:24孔板细胞密度2×10~4/孔,FCCP浓度为2μM。MTT结果中对人肝细胞增殖有显著抑制作用的APAP(4、8、16 mM)能干扰人肝细胞的线粒体有氧呼吸,其中4、8 mM APAP有降低肝细胞基础呼吸和ATP生成的趋势,16 mM APAP能显著抑制肝细胞的基础呼吸、最大呼吸和ATP合成。结论高剂量APAP(16 mM)在人肝细胞活细胞体系中能够显著抑制细胞能量代谢,对活细胞线粒体有氧呼吸显示显著毒性作用。

关键词：对乙酰氨基酚 seahorse XFe24 线粒体有氧呼吸肝毒性

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