白藜芦醇对LPS诱导HUVEC细胞线粒体跨膜电位影响研究

目的:探讨白藜芦醇对LPS刺激HUVEC细胞活性及线粒体跨膜电位影响。方法:体外培养HUVEC,随机分为LPS组、阴性组、白藜芦醇160、80、40、20μg·l-1剂量组。其中,LPS组与白藜芦醇各组分别以1μg·ml-1 LPS刺激细胞24h后,采用CCK-8法检测细胞活力,荧光显微镜下观察罗丹明染色后细胞线粒体跨膜电位。结果:白藜芦醇各组均能提高LPS诱导的HUVEC细胞存活率,提高细胞线粒体跨膜电位,与模型组比较有统计学差异(P0.05)。结论:白藜芦醇能提高LPS刺激HUVEC细胞活性,提高细胞线粒体跨膜电位。     

白藜芦醇缓解低氧/复氧诱导的TCMK1细胞线粒体自噬

目的 研究白藜芦醇(Res)对低氧/复氧(H/R)导致小鼠肾小管上皮细胞系TCMK1内线粒体功能及线粒体自噬的影响。方法 体外建立H/R细胞模型,经Res或(和)线粒体自噬抑制剂1 (Mdivi-1)预处理TMCK1细胞2 h,采用CCK8法检测TCMK1细胞存活率;试剂盒法检测钙离子超负荷情况;流式细胞测量术测定线粒体膜电位(MMP)变化和活性氧(ROS)含量;Western blot检测线粒体融合蛋白(OPA)、分裂蛋白(DRP)以及凋亡相关蛋白cleaved caspase-3,Bax, Bcl2表达;免疫荧光检测线粒体外膜转位酶(TOMM20)和自噬溶酶体相关膜蛋白2(LAMP2)共定位情况。结果 与对照组相比,H/R组TCMK1细胞存活率显著减低(P<0.05),细胞内钙离子、ROS及MMP含量显著减低(P<0.05),OPA表达显著减低(P<0.05),同时DRP表达显著增高(P<0.05),线粒体与LAMP2共定位的细胞数目显著减少;与H/R组相比,10μmol/L Res可以显著增加TMCK1细胞的存活率(P<0.05),抑制细胞内ROS的...     

基于网络药理学探讨白藜芦醇抗衰老的作用机制

目的:运用网络药理学方法探寻白藜芦醇抗衰老的关键靶点及分子作用机制.方法:通过PharmMapper数据库查找白藜芦醇的作用靶点;采用GeneCards数据库搜索衰老的相关蛋白.将白藜芦醇作用靶点和疾病靶点取交集,即为白藜芦醇抗衰老潜在靶点.采用STRING数据库找出靶点的作用关系,以DAVID数据库分析结果,Cyto...     

白藜芦醇干预运动性疲劳模型大鼠线粒体动力学的变化

背景：白藜芦醇是一种存在于葡萄、花生、藜芦等植物中的多酚化合物,是健康食物增补剂的一种。已有研究表明补充白藜芦醇能缓解运动性疲劳,但其具体机制仍不明确,此次研究通过线粒体动力学角度对其机制进行研究。目的：探讨补充白藜芦醇对运动性疲劳大鼠线粒体动力学的影响。方法：48只SD雄性大鼠随机分为空白对照组、白藜芦醇组、运动组和运动+白藜芦醇组,每组12只。运动+白藜芦醇组与运动组同时进行6周负重5%游泳训练,60 min/次,每周6 d。运动+白藜芦醇组在运动后1 h给予白藜芦醇50 mg/kg灌胃,白藜芦醇组只给予白藜芦醇50 mg/kg灌胃,空白对照组与运动组正常饲养,每天同体积溶剂灌胃。末次运动后24 h取材,测定血浆中尿素氮、丙二醛浓度和超氧化物歧化酶活性,骨骼肌中线粒体融合相关因子线粒体融合蛋白1、线粒体融合蛋白2、视神经萎缩蛋白1和分裂相关因子线粒体动力相关蛋白1、线粒体分裂蛋白1的基因表达。结果与结论：(1)与空白对照组相比,运动组血浆中尿素氮、丙二醛浓度明显升高(P <0.05,P <0.05),超氧化物歧化酶活性明显降低(P <0.05);与运动组相比,运动...     

白藜芦醇诱导大鼠肝癌细胞凋亡的作用

目的:探讨白藜芦醇(Res)对大鼠肝癌细胞CBRH7919的作用.方法:采用SRB细胞活性检测法检测白藜芦醇对CBRH7919细胞活性的影响;用流式细胞仪检测CBRH7919对细胞周期分布、凋亡、线粒体膜电位的影响.结果:白藜芦醇作用于CBRH7919细胞24、48和72h后,SRB检测细胞活性有时间和浓度依赖性,CBRH7919细胞凋亡率增加,细胞周期阻断于G0～G1期,细胞线粒体膜电位降低.结论:白黎芦醇对大鼠肝癌细胞CBRH7919有明显的抑制作用,可能是通过线粒体凋亡途径诱导细胞的凋亡.     

白藜芦醇诱导KG-1细胞凋亡作用机制的初步研究

通过观察白藜芦醇诱导人急性白血病细胞株KG-1凋亡的作用,检测bcl-2、caspase-3的表达,探讨白藜芦醇诱导白血病细胞凋亡的作用机制。采用噻唑蓝(MTT)比色法分析细胞生长状态;瑞-吉染色、透射电镜观察KG-1细胞凋亡的形态学变化;流式细胞术(FCM)测定细胞凋亡与周期分布;半定量RT-PCR检测bcl-2 mRNA、caspase-3 mRNA表达水平。结果显示白藜芦醇可明显抑制KG-1细胞增殖(P<0.01),与对照组相比,实验组使细胞发生S期阻滞(P<0.01),促进细胞凋亡(P<0.01),使bcl-2的表达下调,上调caspase-3的表达(P<0.05)。结论:白藜芦醇能诱导KG-1细胞凋亡,其机制可能与下调bcl-2、上调caspase-3表达水平有关。     

白藜芦醇对D-半乳糖诱导的亚急性衰老小鼠海马神经元的保护作用

目的 探讨白藜芦醇对亚急性衰老小鼠海马神经细胞的保护作用.方法 D-半乳糖颈背部皮下注射制备亚急性衰老小鼠模型,设正常对照组、对照组、高低剂量处理组和阳性对照组.第3周始,处理组分别给予白藜芦醇15、45mg/kg(低、高剂量)灌胃,对照组给予生理盐水或DMSO溶液灌胃.8周后,检测小鼠血清、脑组织中超氧化物歧化酶(SOD)活性和丙二醛(MDA)含量.制作石蜡切片,尼氏染色观察小鼠海马CA1区形态结构,免疫组化检测P53蛋白表达,TUNEL法检测细胞凋亡.结果 各组小鼠海马组织形态结构无明显变化;与对照组相比,白藜芦醇45 mg/kg体重组小鼠血清和脑组织中SOD活性明显升高,MDA含量明显下降;白藜芦醇45 mg/kg体重组小鼠海马CA1区P53蛋白表达阳性细胞较对照组小鼠减少,但各组均无明显的凋亡细胞.结论 白藜芦醇能增强亚急性衰老小鼠的抗氧化功能,降低亚急性衰老小鼠海马CA1区神经元P53蛋白的表达.     

白藜芦醇减轻脂多糖诱导的小鼠急性肺损伤

目的观察白藜芦醇在脂多糖(LPS)诱导的急性肺损伤(ALI)小鼠中的作用,以及白藜芦醇对小鼠肺组织NOD样受体蛋白3(NLRP3)表达的影响。方法将小鼠分为对照组,ALI模型组(经气管滴注5 mg/kg的LPS建立小鼠ALI模型),白藜芦醇干预组(经腹腔注射30 mg/kg白藜芦醇,2 h后,经气管滴注5 mg/kg的LPS)。检测小鼠呼吸功能;HE染色及病理评分观察小鼠肺组织形态的变化;检测支气管肺泡灌洗液(BALF)中总蛋白、总细胞数及中性粒细胞数目,并观察中性粒细胞的活化; ELISA检测BALF中IL-1β和IL-18的蛋白水平;real-time PCR检测小鼠肺组织IL-1β、IL-18、nlrp3、asc及pro-caspase-1 mRNA的表达;Western blot检测肺组织IκB的蛋白表达。结果白藜芦醇可改善ALI小鼠的呼吸功能;减轻LPS诱导的肺部病理损伤;降低ALI小鼠BALF中IL-1β和IL-18的蛋白水平(P0.05);减少ALI小鼠肺组织NLRP3、ASC、pro-caspase-1的表达,增加IκB的蛋白表达(P0.05)。结论白藜芦醇可能抑制NLPR3炎性反应小体活化后产物的表达,最终可减轻LPS诱导的小鼠ALI。     

白藜芦醇减轻脂多糖诱导的小鼠心脏损伤

目的 研究白藜芦醇(RES)可否减轻脂多糖(LPS)诱导的小鼠心脏损伤。方法 将小鼠随机分为对照组、LPS组(腹腔注射LPS 7.5 mg/kg)、RES低(20 mg/kg)和高剂量(60 mg/kg)干预组。超声检测心功能;心脏组织切片HE及TUNEL染色观察小鼠心脏组织结构及细胞凋亡;Western blot检测凋亡相关蛋白Bax和Bcl-2的表达;微板法检测血清中CK-MB和LDH水平;ELISA法检测血清中促炎细胞因子IL-1β、IL-6及TNF-α水平;用试剂盒检测心脏组织匀浆中SOD活力和MDA含量。结果 与对照组相比,LPS组小鼠心功能下降(P<0.05);组织结构改变(P<0.05);CK-MB、LDH、IL-1β、IL-6及TNF-α释放增加(P<0.05);SOD活性下降、MDA含量增加(P<0.05);同时心肌凋亡细胞数量增多及Bax与Bcl-2比值升高(P<0.05),RES干预可明显逆转这些改变(P<0.05),且呈剂量依赖性。结论 RES可通过抗炎、抗氧化及抗凋亡作用减轻LPS诱导的心脏损伤。     

白藜芦醇诱导人宫颈癌细胞凋亡的作用

目的观察白藜芦醇对人宫颈癌Hela细胞凋亡的影响。方法采用TUNEL法检测不同浓度的白藜芦醇诱导体外培养的Hela细胞凋亡情况。结果浓度大于50μmol／L的白藜芦醇处理Hela细胞后,细胞凋亡特征明显可见,并呈时间和剂量依赖关系。结论白藜芦醇对人宫颈癌Hela细胞有明显的促凋亡作用。     

Regulation of mitochondrial biogenesis by thyroid hormone

Thyroid hormone (T3) has a profound effect on mitochondrial biogenesis. T3-regulated gene expression is mediated by thyroid hormone receptor (TR) binding to thyroid hormone response elements (TREs). In concert with the action of various coactivators and corepressors this interaction leads to a modulation of the chromatin structure and subsequently to a modulation of gene expression of adjacent target genes. However, as numerous genes are endogenously regulated by T3, and a TRE appears to be absent in their regulatory elements, a TR-independent pathway of T3-mediated gene regulation is likely. In this review, we discuss the direct mechanisms of TR-dependent regulation of gene expression on the nuclear and mitochondrial genome by T3. We also summarise recent observations on an indirect mechanism of T3 action via intermediate factor(s). We discuss the regulation of nuclear respiratory factor 1 (NRF-1) and peroxisome proliferator-activated receptor gamma coactivator 1 alpha (PGC-1alpha) by T3, suggesting NRF-1 and PGC-1alpha as attractive candidates for an intermediate factor of T3 action in vivo.     

高脂血症小鼠脑缺血再灌注后脑组织线粒体功能降低

脑缺血再灌注(ischemia reperfusion,IR)损伤与细胞能量代谢障碍密切相关,细胞内线粒体酶是保证细胞产生ATP的重要因素。线粒体酶活性下降可直接干扰细胞呼吸链的电子传递,阻碍三羧酸循环的进程,影响细胞能量代谢。本实验观察了高脂血症小鼠脑IR后30 d内不同时期脑海马组织琥珀酸脱氢酶(SDH)和细胞色素氧化酶(CCO)活力以及线粒体ATP合成速率的变化,旨在探讨能量代谢障碍在脑IR损伤中所起的作用。1材料与方法1·1仪器和试剂:日本产UV-120-C2型分光光度计,美国产Waters-840型高效液相色谱仪。SDH、CCO活力测定试剂盒、ATP生成速…     

SIRT3 regulates mitochondrial biogenesis in aging-related diseases

Sirtuin 3 (SIRT3), the main family member of mitochondrial deacetylase, targets the majority of substrates controlling mitochondrial biogenesis via lysine deacetylation and modulates important cellular functions such as energy metabolism, reactive oxygen species production and clearance, oxidative stress, and aging. Deletion of SIRT3 has a deleterious effect on mitochondrial biogenesis, thus leading to the defect in mitochondrial function and insufficient ATP production. Imbalance of mitochondrial dynamics leads to excessive mitochondrial biogenesis, dampening mitochondrial function. Mitochondrial dysfunction plays an important role in several diseases related to aging, such as cardiovascular disease, cancer and neurodegenerative diseases. Peroxisome proliferator-activated receptor gamma coactivator 1-alpha (PGC1α) launches mitochondrial biogenesis through activating nuclear respiratory factors. These factors act on genes, transcribing and translating mitochondrial DNA to generate new mitochondria. PGC1α builds a bridge between SIRT3 and mitochondrial biogenesis. This review described the involvement of SIRT3 and mitochondrial dynamics, particularly mitochondrial biogenesis in aging-related diseases, and further illustrated the role of the signaling events between SIRT3 and mitochondrial biogenesis in the pathological process of aging-related diseases.     

Drug-induced increased mitochondrial biogenesis in a liver biopsy

Oncocytic changes seen in hepatocytes in patients receiving highly active antiretroviral therapy (HAART) are a result of mitochondrial damage. This is the first report that provides the electron microscopy illustration of mitochondrial proliferation as a result of the HAART drug Stavudine (Zerit) hepatotoxicity. The drug's effect on mitochondrial DNA replication leads to depleted mitochondrial-encoded proteins and configurational defects of the mitochondrial inner membrane leading to reduced and abnormal cristae, which house the electron transport chain and elementary bodies. This results in a decrease in the NAD/NADH ratio and reduces oxidative phosphorylation. The shift in the NAD/NADH ratio decreases the rate of fatty acid beta oxidation and oxidation of pyruvate by the Krebs cycle. Decreased oxidation of pyruvate drives it into an alternative pathway to form lactate leading to lactic acidosis. This mitochondrial dysfunction results in a compensatory increase in mitochondrial biogenesis, which results in oncocytic changes of the hepatocytes.     

The role of AMP-activated protein kinase in mitochondrial biogenesis

While it has been known for more than 75 years that physical activity is associated with increased mitochondrial content in muscle, the molecular mechanism for this adaptive process has only recently been elucidated. This brief review examines existing studies that have identified AMPK-activated protein kinase (AMPK) and several other key regulators of mitochondrial biogenesis, including peroxisome proliferator-activated receptor-γ coactivator-1α and -1β, calcium/calmodulin-dependent protein kinase IV, and nitric oxide. In addition, the potential role of mitochondrial dysfunction in the pathogenesis of insulin resistance associated with ageing and type 2 diabetes mellitus is also discussed.