Hepatic Mitochondrial Respiratory Activation, Depolarization and Recovery After Acute Ethanol
急性乙醇后肝线粒体呼吸激活、去极化和恢复
基本信息
- 批准号:10607356
- 负责人:
- 金额:$ 4.42万
- 依托单位:
- 依托单位国家:美国
- 项目类别:
- 财政年份:2022
- 资助国家:美国
- 起止时间:2022-09-01 至 2025-08-31
- 项目状态:未结题
- 来源:
- 关键词:AccidentsAcetaldehydeAcetatesAcuteAddressAdenosineAlcohol abuseAlcohol consumptionAlcohol dependenceAlcoholic Liver DiseasesBehavior DisordersBiogenesisCardiovascular DiseasesCause of DeathCessation of lifeChronicCirrhosisConfocal MicroscopyDangerousnessDevelopmentDiseaseDsRedDyesEthanolEthanol MetabolismEventFibrosisFluorescenceGenus HippocampusHealthHepaticHepatocyteHourIn VitroIndividualKnowledgeLabelLaboratoriesLeadLiverLiver FailureLiver FibrosisLiver MitochondriaLiver diseasesMalignant NeoplasmsMedicalMembrane PotentialsMental disordersMentorsMetabolicMitochondriaMitochondrial DNAMitochondrial Proton-Translocating ATPasesMolecularMusNucleotidesOxygen ConsumptionOxygen saturation measurementPatternPermeabilityPersonsProcessProductivityProteinsProtonsReactive Oxygen SpeciesRecoveryResearchRiskRoleSerumSteatohepatitisTimeUCP2 proteinUnited StatesWorkplacealcohol responsecostcost estimateextracellularfluorophorein vivoinhibitorliver inflammationmitochondrial dysfunctionmitochondrial membranemitochondrial metabolismmitochondrial permeability transition poremultiphoton microscopynovel therapeuticsoxidationpreventrespiratoryresponse
项目摘要
ABSTRACT
Liver failure is a major cause of death worldwide. Hepatic mitochondrial depolarization (mtDepo) is one of
the earliest responses to ethanol and likely is the first in a chain of events leading to subsequent liver disease.
Initially, mtDepo in response to ethanol facilitates more rapid two-step oxidation of ethanol and its toxic metabolite
acetaldehyde (AcAld) to acetate. I hypothesize that mtDepo underlies a swift increase in alcohol
metabolism (SIAM) after acute ethanol administration that is an adaptive response to eliminate ethanol
more rapidly. Chronically, the response becomes maladaptive leading to disordered mitophagy and hepatic
inflammation and fibrosis. Furthermore, I hypothesize that mtDepo after EtOH is brought about via mitochondrial
uncoupling due to proton leaks through either the adenosine nucleotide translocator (ANT1/2), the mitochondrial
F1FO-ATP synthase, uncoupling proteins (UCPs) or mitochondrial permeability transition (PT) pores. Since
mtDepo leads to elimination of mitochondria by mitophagy, I also hypothesize that recovery from mtDepo
after ethanol involves mitochondrial biogenesis. In two Specific Aims, I will: 1) Characterize hepatocyte
mitochondrial oxygen consumption rate (OCR) in relation to mitochondrial depolarization and
repolarization after acute ethanol. I will assess time-dependent changes in OCR and mitochondrial membrane
potential (ΔΨ) of hepatocytes freshly isolated from ethanol-treated and untreated mice using Seahorse
extracellular flux analysis, Hansatech oximetry, and confocal microscopy of fluorescent ΔΨ indicators. By
injecting in vivo prior to hepatocyte isolation MitoTracker dyes, fluorophores that label only polarized
mitochondria, I will identify hepatocytes having undergone mtDepo in vivo in relation to mtDepo and
repolarization in vitro. To address my mechanistic hypotheses, I will assess how specific inhibitors of ANT, ATP
synthase, UCPs, and PT pores reverse/prevent ethanol-induced mtDepo and increased OCR. 2) Assess the
role of mitochondrial biogenesis and mitophagy in recovery from mtDepo after acute ethanol. As mice
metabolically eliminate ethanol, hepatocytes recover from mtDepo. In Mitotimer mice, newly synthesized DsRed-
E5 fluoresces green but over time (12-24 h) shifts irreversibly to red fluorescence. If repolarization of preexisting
mitochondria occurs after ethanol, then Mitotimer fluorescence should be predominantly red in mitochondria
recovering from mtDepo. If repolarization results from mitochondrial biogenesis, then recovering mitochondria
will fluoresce green. I expect that repolarization of preexisting mitochondria and biogenesis of new mitochondria
will both contribute to recovery from mtDepo after acute ethanol. Together, Aims 1 and 2 will characterize key
mechanisms in the hepatic mitochondrial response to ethanol, as well as identify the basis for recovery. This
fundamental knowledge could lead to development of new therapeutics to treat and prevent alcoholic liver
disease.
摘要
肝衰竭是世界范围内的主要死亡原因。肝线粒体去极化(mtDepo)是
对乙醇的最早反应,可能是导致随后肝脏疾病的一连串事件中的第一个。
最初,mtDepo响应于乙醇促进乙醇及其有毒代谢物的更快速的两步氧化
乙醛(AcAld)转化为乙酸盐。我假设mtDepo是酒精迅速增加的基础
急性乙醇给药后的SIAM代谢(SIAM)是消除乙醇的适应性反应
更快。慢性时,这种反应变得适应不良,导致线粒体自噬紊乱和肝细胞凋亡。
炎症和纤维化。此外,我假设EtOH后的mtDepo是通过线粒体
解偶联是由于质子通过腺苷酸转运蛋白(ANT 1/2)、线粒体膜蛋白(线粒体膜蛋白)
F1 FO-ATP合酶、解偶联蛋白(UCP)或线粒体通透性转换(PT)孔。以来
mtDepo通过线粒体自噬导致线粒体消除,我还假设从mtDepo中恢复
因为乙醇涉及线粒体生物合成。在两个具体目标中,我将:1)表征肝细胞
线粒体耗氧率(OCR)与线粒体去极化和
急性乙醇后复极。我将评估OCR和线粒体膜的时间依赖性变化,
使用Seahorse从乙醇处理和未处理小鼠中新鲜分离的肝细胞的电位(Δ λ)
细胞外通量分析、Hansatech血氧测定法和荧光Δ λ指示剂的共聚焦显微镜。通过
在肝细胞分离之前体内注射MitoTracker染料,仅标记极化的荧光团
线粒体,我将鉴定与mtDepo相关的体内经历mtDepo的肝细胞,
体外复极化。为了解决我的机制假设,我将评估ANT的特异性抑制剂ATP
合成酶、UCP和PT孔逆转/防止乙醇诱导的mtDepo和增加的OCR。2)评估
线粒体生物发生和线粒体自噬在急性乙醇后从mtDepo恢复中的作用。诸如鼠标
代谢消除乙醇,肝细胞从mtDepo中恢复。在线粒体小鼠中,新合成的DsRed-
E5发出绿色荧光,但随着时间的推移(12-24小时)不可逆地转变为红色荧光。如果先前存在的复极化
线粒体在乙醇后发生,那么线粒体中Mitotimer荧光应该主要是红色的
从mtDepo恢复过来如果复极化是线粒体生物发生的结果,
会发出绿色荧光。我认为先存线粒体的复极化和新线粒体的生物合成
都将有助于急性乙醇后从mtDepo中恢复。目标1和目标2将共同构成
肝脏线粒体对乙醇的反应机制,以及确定恢复的基础。这
基础知识可能导致开发新的治疗方法来治疗和预防酒精性肝
疾病
项目成果
期刊论文数量(0)
专著数量(0)
科研奖励数量(0)
会议论文数量(0)
专利数量(0)
数据更新时间:{{ journalArticles.updateTime }}
{{
item.title }}
{{ item.translation_title }}
- DOI:
{{ item.doi }} - 发表时间:
{{ item.publish_year }} - 期刊:
- 影响因子:{{ item.factor }}
- 作者:
{{ item.authors }} - 通讯作者:
{{ item.author }}
数据更新时间:{{ journalArticles.updateTime }}
{{ item.title }}
- 作者:
{{ item.author }}
数据更新时间:{{ monograph.updateTime }}
{{ item.title }}
- 作者:
{{ item.author }}
数据更新时间:{{ sciAawards.updateTime }}
{{ item.title }}
- 作者:
{{ item.author }}
数据更新时间:{{ conferencePapers.updateTime }}
{{ item.title }}
- 作者:
{{ item.author }}
数据更新时间:{{ patent.updateTime }}
Matthew Savoca其他文献
Matthew Savoca的其他文献
{{
item.title }}
{{ item.translation_title }}
- DOI:
{{ item.doi }} - 发表时间:
{{ item.publish_year }} - 期刊:
- 影响因子:{{ item.factor }}
- 作者:
{{ item.authors }} - 通讯作者:
{{ item.author }}
{{ truncateString('Matthew Savoca', 18)}}的其他基金
Hepatic Mitochondrial Respiratory Activation, Depolarization and Recovery After Acute Ethanol
急性乙醇后肝线粒体呼吸激活、去极化和恢复
- 批准号:
10748288 - 财政年份:2022
- 资助金额:
$ 4.42万 - 项目类别:
相似海外基金
An enzyme-based assay for the detection of acetaldehyde-protein adducts
用于检测乙醛-蛋白质加合物的酶测定法
- 批准号:
10760201 - 财政年份:2023
- 资助金额:
$ 4.42万 - 项目类别:
Dissecting the role of acetaldehyde in oral carcinogenesis
剖析乙醛在口腔癌发生中的作用
- 批准号:
10345780 - 财政年份:2022
- 资助金额:
$ 4.42万 - 项目类别:
Dissecting the role of acetaldehyde in oral carcinogenesis
剖析乙醛在口腔癌发生中的作用
- 批准号:
10706454 - 财政年份:2022
- 资助金额:
$ 4.42万 - 项目类别:
Identification of mutation patterns induced by acetaldehyde exposure
乙醛暴露引起的突变模式的鉴定
- 批准号:
20K17047 - 财政年份:2020
- 资助金额:
$ 4.42万 - 项目类别:
Grant-in-Aid for Early-Career Scientists
RUI: Collaborative: Cycling of ethanol and acetaldehyde in coastal waters
RUI:合作:沿海水域乙醇和乙醛的循环
- 批准号:
2022184 - 财政年份:2020
- 资助金额:
$ 4.42万 - 项目类别:
Standard Grant
Reconsideration of the drinking habit of alcoholic liver disease patients from the viewpoint of acetaldehyde-derived advanced glycation end products
从乙醛衍生晚期糖基化终末产物角度重新思考酒精性肝病患者饮酒习惯
- 批准号:
19K11803 - 财政年份:2019
- 资助金额:
$ 4.42万 - 项目类别:
Grant-in-Aid for Scientific Research (C)
Pathogenic Role of Malondialdehyde-Acetaldehyde Adducts in Rheumatoid Arthritis
丙二醛-乙醛加合物在类风湿性关节炎中的致病作用
- 批准号:
10421254 - 财政年份:2019
- 资助金额:
$ 4.42万 - 项目类别:
Pathogenic Role of Malondialdehyde-Acetaldehyde Adducts in Rheumatoid Arthritis
丙二醛-乙醛加合物在类风湿性关节炎中的致病作用
- 批准号:
10045500 - 财政年份:2019
- 资助金额:
$ 4.42万 - 项目类别:
Pathogenic Role of Malondialdehyde-Acetaldehyde Adducts in Rheumatoid Arthritis
丙二醛-乙醛加合物在类风湿性关节炎中的致病作用
- 批准号:
10516090 - 财政年份:2019
- 资助金额:
$ 4.42万 - 项目类别:
Analysis of the molecular mechanism to repair acetaldehyde-induced DNA damage
修复乙醛所致DNA损伤的分子机制分析
- 批准号:
17K17846 - 财政年份:2017
- 资助金额:
$ 4.42万 - 项目类别:
Grant-in-Aid for Young Scientists (B)