益气解毒方对糖尿病肾病大鼠肾小管上皮细胞线粒体自噬作用机制

目的:研究益气解毒方对糖尿病肾病(diabetic nephropathy,DN)大鼠肾小管上皮细胞线粒体自噬作用机制。方法:采用DN大鼠模型,观察益气解毒方对DN的治疗作用。采用苏木素伊红(HE)染色法观察给药后DN大鼠肾脏病理变化,透射电镜观察DN大鼠肾小管上皮细胞线粒体自噬体情况。采用蛋白免疫印迹法检测肾组织中微管相关蛋白1轻链3Ⅱ(microtubule-associatedprotein1lightchain 3Ⅱ,MAP1LC3-Ⅱ,LC3-Ⅱ),自噬标志物p62蛋白(sequestosome-1,p62),促凋亡蛋白Nix(Bcl-2/E1B 19 k Da-interacting protein 3-like,Nix)的表达,并应用实时荧光定量PCR测定Nix mRNA的表达。结果:与模型组比较,益气解毒方能够增加DN大鼠体重,改善DN大鼠血尿素氮(BUN),血肌酐(SCr),血糖(GLU),甘油三酯(TG)及总胆固醇(CHOL),白蛋白(Alb)等血、尿生化指标(P0.01),并且能抑制由DN导致的肾小管上皮细胞线粒体过度自噬,显著降低LC3-Ⅱ,p62,Nix蛋白的表达(P0.01),同时降低Nix mRNA的表达(P0.01)。结论:益气解毒方对DN有较好的治疗效果,这可能与其调节DN肾小管上皮细胞线粒体自噬紊乱有关。     

Targeting autophagy and mitophagy for mitochondrial diseases treatment

Introduction: Mitochondrial diseases are a group of rare genetic diseases with complex and heterogeneous origins which manifest a great variety of phenotypes. Disruption of the oxidative phosphorylation system is the main cause of pathogenicity in mitochondrial diseases since it causes accumulation of reactive oxygen species (ROS) and ATP depletion.

Areas covered: Current evidences support the main protective role of autophagy and mitophagy in mitochondrial diseases and other diseases associated with mitochondrial dysfunction.

Expert Opinion: The use of autophagy and/or mitophagy inducers may allow a novel strategy for improving mitochondrial function for both mitochondrial diseases and other diseases with altered mitochondrial metabolism. However, a deeper investigation of the molecular mechanisms behind mitophagy and mitochondrial biogenesis is needed in order to safely modulate these processes. In the coming years, we will also see an increase in awareness of mitochondrial dynamics modulation that will allow the therapeutic use of new drugs for improving mitochondrial function in a great variety of mitochondrial disorders.     

大黄-桃仁药对调控子宫腺肌病在位内膜线粒体稳态的机制研究

目的 探索大黄-桃仁药对对子宫腺肌病（adenomyosisi,AM）在位内膜线粒体稳态的调控机制。方法 同种异体垂体移植法构建AM小鼠模型,光镜及透射电镜（TEM）观察大黄-桃仁药对治疗前后子宫组织病理及超微结构改变;免疫荧光检测线粒体自噬通路PTEN诱导的推定激酶1(PTEN-induced putative kinase 1,PINK1)/帕金森蛋白（Parkinson protein,Parkin）蛋白表达。培养人永生化子宫内膜异位症在位内膜间质细胞hEM15A,观察大黄-桃仁药对处理前后细胞增殖能力、活性氧（reactive oxygen species,ROS）、抗氧化酶系、线粒体膜电位、线粒体自噬小体、线粒体自噬蛋白的变化。结果 病理切片显示,经大黄-桃仁药对治疗后,中、重度的浸润比例较模型组减少。TEM观察发现模型组子宫内膜上皮细胞及间质细胞线粒体肿胀、线粒体脊消失,部分形成线粒体自噬体;经大黄-桃仁药对治疗后子宫内膜上皮细胞及间质细胞线粒体数量增加,无明显肿胀、线粒体脊清晰可见。与对照组比较,造模后子宫内膜层PINK1、Parkin表达显著上调（P<0.001）...     

保肾通络方含药血清调节线粒体自噬对高糖诱导下足细胞氧化损伤的影响

目的 观察保肾通络方含药血清对高糖培养下足细胞氧化损伤的影响,并对保肾通络方的作用机制进行探讨。方法 以体外培养的肾小球足细胞为研究对象,给予高浓度葡萄糖刺激,采用CCK-8法分析足细胞活力以确定最佳模型及给药浓度,分为正常对照组、高糖组、保肾通络方组,采用鬼笔环肽进行细胞骨架染色,Western blotting法检测足细胞裂孔膜蛋白nephrin,自噬相关蛋白5(autophagy-related protein 5,ATG5)、ATG7、微管相关蛋白1轻链3(microtubule-associated protein 1 light chain 3,LC3)的表达,流式细胞术检测足细胞活性氧(reactive oxygen species,ROS)及线粒体膜电位,免疫荧光共标LC3B及线粒体,观察各组足细胞内线粒体自噬泡的生成情况。结果 30 mmol/L高糖干预48 h可显著降低足细胞活力(P＜0.05),保肾通络方含药血清可使高糖诱导的足细胞活力明显提升(P＜0.01);与正常对照组相比,高糖组足细胞骨架褶皱呈多边形,应力纤维束被破坏,ROS生成显著增多(P＜0.01),足细胞nephrin、ATG5、ATG7表达显著降低,LC3Ⅱ/LC3Ⅰ比值降低(P＜0.05),LC3B与线粒体共标减少,线粒体膜电位下降(P＜0.01);与高糖组相比,保肾通络方组足细胞骨架改善,ROS生成减少(P＜0.01),nephrin、ATG5、ATG7表达显著增加(P＜0.05),LC3Ⅱ/LC3Ⅰ比值上升(P＜0.05),LC3B与线粒体荧光共标增多,线粒体膜电位升高(P＜0.01)。结论 保肾通络方含药血清能够减轻高糖诱导的足细胞氧化损伤,其作用可能与改善足细胞线粒体自噬有关。     

Human Cell Organelles in SARS-CoV-2 Infection: An Up-to-Date Overview

Since the end of 2019, the whole world has been struggling with the life-threatening pandemic amongst all age groups and geographic areas caused by Severe Acute Respiratory Syndrome Coronavirus (SARS-CoV-2). The Coronavirus Disease 2019 (COVID-19) pandemic, which has led to more than 468 million cases and over 6 million deaths reported worldwide (as of 20 March 2022), is one of the greatest threats to human health in history. Meanwhile, the lack of specific and irresistible treatment modalities provoked concentrated efforts in scientists around the world. Various mechanisms of cell entry and cellular dysfunction were initially proclaimed. Especially, mitochondria and cell membrane are crucial for the course of infection. The SARS-CoV-2 invasion depends on angiotensin converting enzyme 2 (ACE2), transmembrane serine protease 2 (TMPRSS2), and cluster of differentiation 147 (CD147), expressed on host cells. Moreover, in this narrative review, we aim to discuss other cell organelles targeted by SARS-CoV-2. Lastly, we briefly summarize the studies on various drugs.     

基于线粒体质量控制的荭草苷抗心肌缺血/再灌注损伤的机制研究

目的探讨荭草苷对心肌缺血/再灌注损伤(MIRI)大鼠心肌细胞内线粒体质量的调控作用。方法 75只SD雄性大鼠随机分为假手术组、模型组、不同剂量(1.0、2.0、4.0 mg/kg)荭草苷预处理组。除假手术组外,其余各组大鼠均通过结扎冠状动脉左前降支致缺血30 min,再灌注120 min,制备MIRI模型。酶标仪检测线粒体膜电位(MMP)和线粒体通透性转换孔(mPTP)开放程度,分光光度法测定线粒体呼吸链酶复合物Ⅰ~Ⅳ活性,TUNEL法检测心肌细胞凋亡水平,Western blotting法检测心肌细胞内凋亡与线粒体自噬相关蛋白的表达水平,免疫共沉淀法检测心肌细胞内Parkin与p62结合程度。结果荭草苷可显著恢复MIRI诱导的MMP、m PTP开放阈值和粒体呼吸链酶复合物Ⅰ~Ⅳ活性降低,减少心肌细胞凋亡;并能抑制心肌细胞内凋亡及线粒体自噬相关蛋白表达水平,减弱Parkin与p62的结合程度。结论荭草苷对MIRI大鼠心肌细胞内线粒体具有明显保护作用,其机制可能通过Parkin依赖性和Parkin非依赖性信号通路抑制心肌细胞内线粒体自噬过度激活有关。     

丹皮酚对大鼠心肌梗死后心脏功能的改善及其对Pink1/Parkin的影响＊

目的 检测丹皮酚（paeonol）对大鼠心梗后心肌重构和心脏功能的改善作用并探究其作用机制。方法 采用SD大鼠和人心肌细胞H9C2分别建立体内心肌梗死动物模型和体外心肌细胞缺氧细胞模型。Western blot检测Pink1、Parkin、LC3Ⅰ、LC3Ⅱ、Beclin-1和Mfn2的表达水平。本研究通过ELISA试剂盒检测心肌损伤标志物乳酸脱氢酶（LDH）和肌酸激酶同工酶（CK-MB）在血浆中的含量。细胞凋亡率采用TUNEL检测。使用超声心动图和生物机能采集系统检测大鼠心脏功能和心室重构情况。结果 丹皮酚减少了心肌损伤标志物LDH和CK-MB的释放，并减弱了H9C2细胞凋亡。丹皮酚激活了Pink1/Parkin信号通路，并因此增强了线粒体自噬。进一步的研究表明，丹皮酚显著减轻了SD大鼠心梗后心室重构情况并改善了心脏功能。结论 丹皮酚通过激活Pink1/Parkin信号通路促进线粒体自噬，进而减轻了心肌细胞损伤，改善了大鼠心脏功能和心室重构情况。     

浅谈Mfn2对心力衰竭的影响

Mitofusin 2 (Mfn2) 是一种重要的多功能蛋白，在物理和病理条件下参与各种生物过程，包括线粒体融合、网状线粒体接触、线粒体质量控制和细胞凋亡。已发现 Mfn2 功能障碍会导致心血管疾病，例如心力衰竭。在这里，这篇综述主要关注关于 Mfn2 的结构和功能及其在心力衰竭中的作用。     

Melatonin suppresses platelet activation and function against cardiac ischemia/reperfusion injury via PPARγ/FUNDC1/mitophagy pathways

Platelet activation is a major (patho‐) physiological mechanism that underlies ischemia/reperfusion (I/R) injury. In this study, we explored the molecular signals for platelet hyperactivity and investigated the beneficial effects of melatonin on platelet reactivity in response to I/R injury. After reperfusion, peroxisome proliferator‐activated receptor γ (PPARγ) was progressively downregulated in patients with acute myocardial infarction undergoing coronary artery bypass grafting (CABG) surgery and in mice with I/R injury model. Loss of PPARγ was closely associated with FUN14 domain containing 1 (FUNDC1) dephosphorylation and mitophagy activation, leading to increased mitochondrial electron transport chain complex (ETC.) activity, enhanced mitochondrial respiratory function, and elevated ATP production. The improved mitochondrial function strongly contributed to platelet aggregation, spreading, expression of P‐selectin, and final formation of micro‐thromboses, eventually resulting in myocardial dysfunction and microvascular structural destruction. However, melatonin powerfully suppressed platelet activation via restoration of the PPARγ content in platelets, which subsequently blocked FUNDC1‐required mitophagy, mitochondrial energy production, platelet hyperactivity, and cardiac I/R injury. In contrast, genetic ablation of PPARγ in platelet abolished the beneficial effects of melatonin on mitophagy, mitochondrial ATP supply, and platelet activation. Our results lay the foundation for the molecular mechanism of platelet activation in response to I/R injury and highlight that the manipulation of the PPARγ/FUNDC1/mitophagy pathway by melatonin could be a novel strategy for cardioprotection in the setting of cardiac I/R injury.     

Mitochondrial biogenesis and dynamics in the developing and diseased heart

The mitochondrion is a complex organelle that serves essential roles in energy transduction, ATP production, and a myriad of cellular signaling events. A finely tuned regulatory network orchestrates the biogenesis, maintenance, and turnover of mitochondria. The high-capacity mitochondrial system in the heart is regulated in a dynamic way to generate and consume enormous amounts of ATP in order to support the constant pumping function in the context of changing energy demands. This review describes the regulatory circuitry and downstream events involved in mitochondrial biogenesis and its coordination with mitochondrial dynamics in developing and diseased hearts.