腺苷酸活化蛋白激酶激活剂的研究进展

腺苷酸活化蛋白激酶(AMPK)在真核细胞的能量感受与调节中起关键作用,被AMP等其他因素激活后具有加速分解代谢、降低合成代谢的功能,从而稳定人体糖代谢与脂代谢。AMPK激活剂在2型糖尿病等代谢异常的动物模型中体现出较好的治疗效果。对近年来AMPK激活剂的研究进展进行综述,期望对该类药物的研发提供参考。     

腺苷酸活化蛋白激酶在肺部疾病中的作用

腺苷酸活化蛋白激酶(AMP-activated protein kinase,AMPK)是一种广泛参与人体内能量代谢活动的蛋白激酶。新近的研究发现,AMPK可调控多种疾病发生、发展过程中的病理环节。近年来,AMPK与肺部疾病的关系愈发受到人们的重视,目前研究认为AMPK对肺癌、支气管哮喘及肺动脉高压等肺部疾病的治疗具有潜在的作用。该文就AMPK的生物学特性及其对多种肺部疾病的潜在治疗作用进展做一综述。     

腺苷酸活化蛋白激酶及脂肪酸合成酶在乳腺癌合并2型糖尿病患者中表达水平分析

目的研究腺苷酸活化蛋白激酶(AMPK)及脂肪酸合成酶(FASN)在乳腺癌合并2型糖尿病患者中表达水平。方法选取2012年5月至2016年5月入住我院确诊为乳腺癌的女性患者共160例,其中80例乳腺癌患者合并患有2型糖尿病,采用免疫组织化学方法对AMPK和FASN在患者乳腺中的表达水平进行测定和统计分析。结果腺苷酸活化蛋白激酶在乳腺癌患者组中的表达水平高于乳腺癌合并2型糖尿病患者组的表达水平,差异有统计学意义(P<0.05);脂肪酸合成酶在乳腺癌患者组中的表达水平低于乳腺癌合并2型糖尿病患者组的表达水平,差异有统计学意义(P<0.05)。结论 AMPK和FASN可能是肿瘤组织产生和进一步发展过程中的重要影响因子,在此过程中AMPK活性受到抑制,FASN活性得到增强。     

二甲双胍不依赖腺苷酸活化蛋白激酶途径的抗肿瘤机制研究进展

二甲双胍是临床上广泛用来治疗2型糖尿病的口服降糖药。流行病学研究发现,二甲双胍与癌症的预防和治疗有着密切的关系。二甲双胍的抗肿瘤分子机制已成为研究热点。大部分研究认为,该药理作用依赖腺苷酸活化蛋白激酶(AMPK)的激活。近几年,部分学者提出二甲双胍通过非AMPK依赖途径抗肿瘤的机制,可改善胰岛素敏感性,下调Ras相关GTP结合蛋白Rag活性和DNA损伤反应调节基因1的表达,抑制哺乳动物雷帕霉素靶蛋白(mTOR)信号通路,抑制Ras相关C3肉毒杆菌毒素底物活性、细胞分裂周期蛋白42和基质金属蛋白酶的表达等多种途径抑制肿瘤细胞的生长、侵袭和迁移。本综述以此为切入点,综述二甲双胍不依赖AMPK途径的抗肿瘤机制研究进展,为其抗肿瘤作用机制的探讨展示新视角。     

腺苷酸活化蛋白激酶调控相关通路与糖脂代谢异常相关疾病的发病机制研究进展

糖脂代谢异常会造成线粒体功能紊乱,从而导致相关疾病的发生发展.研究表明,腺苷酸活化蛋白激酶(AMPK)是体内重要的能量代谢感受器,激活后的AMPK通过调控多条相关通路参与机体糖脂代谢和能量调节活动,以减轻糖脂代谢紊乱对机体的损害或促进机体相关正常功能的恢复.本文对AMPK调控相关通路在糖脂代谢异常相关疾病中的研究进展进...     

脂联素对体外循环心肌胰岛素抵抗模型犬心肌腺苷酸活化蛋白激酶表达的影响

目的:探讨脂联素(APN)对体外循环(CPB)心肌胰岛素抵抗(IR)模型犬心肌腺苷酸活化蛋白激酶(AMPK)表达的影响。方法:将24只犬随机分为对照组、模型组、APN组(36μg/kg)、AMPK抑制剂组(APN 36μg/kg+AMPK抑制剂复合制剂C 0.5mg/kg),每组6只。所有犬行CPB术,除对照组不给药外,其余各组犬建立CPB心肌IR模型,并于主动脉夹闭后灌注不含药或含相应药物的St.Thomas心脏停搏液。分别在CPB转流前和心缺血60 min后再灌注15、90 min收集冠状静脉窦血、颈动脉血,取左心室心尖部组织,测定并计算心肌葡萄糖摄取率和胰岛素抵抗指数(IRI),监测心肌损伤指标[肌钙蛋白T(c TnT)浓度]和心功能指标[左室收缩压(LVSP)、左室内压最大上升速率(+dp/dt_(max))]变化,检测磷酸化AMPK(p-AMPK)水平。结果:转流前各组犬之间上述指标差异均无统计学意义(P>0.05)。与对照组比较,模型组犬再灌注15、90 min的心肌葡萄糖摄取率、LVSP、+dp/dt_(max)和pAMPK水平均明显降低(P<0.05),IRI和cTnT浓度均明显升高(P<0.05)。与模型组比较,APN组和AMPK抑制剂组犬再灌注15、90 min的心肌葡萄糖摄取率、LVSP、+dp/dt_(max)和p-AMPK水平均明显升高(P<0.05),IRI和cTnT浓度均明显降低(P<0.05),且APN组效果强于AMPK抑制剂组(P<0.05)。结论:APN可促进心肌对葡萄糖的摄取及代谢以助心功能恢复,其可能是通过增强AMPK活性来发挥效应的。     

牛蒡子苷对L6骨骼肌细胞中腺苷酸活化蛋白激酶信号通路的影响

目的：探讨牛蒡子苷在L6骨骼肌细胞中对葡萄糖消耗和腺苷酸活化蛋白激酶（ AMP?activated protein kinase, AMPK）信号通路的调节作用。方法用含2％胎牛血清的1640培养基诱导L6骨骼肌细胞分化,直至细胞生长出肌管,分化完成后的L6骨骼肌细胞用含有不同浓度牛蒡子苷的培养基培养24 h后,采用葡萄糖氧化酶法检测细胞葡萄糖消耗,western blot法检测AMPK的亚单位AMPKα和磷酸化腺苷酸活化蛋白激酶（ p?AMP?activated protein kinase,p?AMPKα）的蛋白表达水平;以real?time PCR法检测过氧化物酶体增生物激活受体γ共激活因子1α（ peroxisome?proliferator?activated receptorγcoactiva?tor?1α,PGC?1α） mRNA表达水平。结果牛蒡子苷浓度为1000 g／L 时,可增加 L6骨骼肌细胞的葡萄糖消耗,差异显著（ P＜0．01）;与对照组相比,牛蒡子苷呈浓度依赖性的增强L6骨骼肌细胞中p?AMPKα蛋白表达水平;并以相同的趋势增强PGC?1α mRNA表达水平。结论牛蒡子苷增加L6骨骼肌细胞的葡萄糖消耗,并激活AMPK／PGC?1α信号通路,具有潜在的改善胰岛素抵抗的作用。     

PPARα、固醇调节元件结合蛋白和腺苷酸活化蛋白激酶在酒精性脂肪肝发病中作用的研究进展

酒精性脂肪肝(AFLD)为酒精性肝损伤的最初症状,对其发生机制的深入研究有助于AFLD的预防和治疗。迄今研究发现,AFLD的发生发展主要与细胞色素P450 2E1、过氧化物酶体增殖物激活受体α(PPARα)、固醇调节元件结合蛋白(SREBP)、腺苷酸活化蛋白激酶(AMPK)、去乙酰化酶1、脂联素和胰岛素等通路相关。研究发现,酒精及代谢产物乙醛直接或间接抑制PPARα和AMPK并激活SREBP蛋白通路,导致肝脂质代谢紊乱、脂肪酸氧化能力降低和脂质堆积,最终形成AFLD。此外,本综述对这3个蛋白作为AFLD治疗靶点的治疗前景予以展望。     

肝宝胶囊对非酒精性脂肪性肝病大鼠的治疗效应及对腺苷酸活化蛋白激酶通路的影响

目的 探讨肝宝胶囊对非酒精性脂肪性肝病(NAFLD)大鼠的治疗效应及对腺苷酸活化蛋白激酶信号通路的影响。方法 40只SD雄性大鼠随机分为4组(n=10):正常组、模型组、药物治疗组(肝宝胶囊低剂量组、肝宝胶囊高剂量组)。正常组以普通饲料喂养,其余3组经高脂饲料喂养建立NAFLD模型,药物治疗组分别予0.54 g/(kg·d)、2.16 g/(kg·d)肝宝胶囊灌胃治疗4周,模型组以蒸馏水灌胃;HE染色评估病理学改变,检测各组血清TC、TG、ALT、AST及肝组织TC、TG含量;ELISA法检测肝组织IL-6、IL-10、FFA、TNF-α含量;Real-time PCR检测肝组织AMPK、SREBP-1c mRNA表达;结果 与模型组比较,药物治疗组均降低大鼠血清TC、TG、ALT、AST水平,降低肝组织FFA、TNF-α、IL-6表达,降低肝脏NAS积分;药物治疗组均升高肝组织IL-10、AMPK mRNA水平,降低SREBP-1c mRNA水平,高剂量组作用均更显著(P<0.05)。结论 肝宝胶囊可能通过激活AMPK通路并抑制SREBP-1c表达改善肝脏脂质代谢及减轻肝细胞炎...     

叉头框家族D3和腺苷酸活化蛋白激酶5与胃癌顺铂耐药机制的研究

目的 研究叉头框家族D3（FOXD3）和腺苷酸活化蛋白激酶5（ARK5）参与胃癌顺铂耐药的机制。方法 将细胞株分为6组：SGC7901组（1组）;SGC7901/DDP组（2组）、SGC7901/DDP-FOXD3组（3组）、SGC7901/DDP-FOXD3-NC组（4组）、SGC7901/DDP-shARK5组（5组）和SGC7901/DDP-shARK5-NC组（6组）。在第3组细胞株中转染过表达FOXD3质粒,第4组中转染FOXD3空白质粒作为对照,在第5组细胞株中转染敲降shARK5质粒,第6组细胞株转染敲降shARK5空白对照质粒。用Western blot法检测FOXD3和ARK5蛋白的表达水平,用噻唑蓝比色法测定顺铂半抑制浓度(IC₅₀)。结果 第1至6组中FOXD3蛋白相对表达量分别为27.15±7.53,22.04±3.92,45.39±5.17,23.39±2.56,21.99±4.74和24.27±5.39,ARK5蛋白相对表达量分别为25.33±5.11,27.56±3.45,27.82±5.40,27.21±5.22,10.33±0.8...     

Screening methods for AMP-activated protein kinase modulators: a patent review

Introduction: AMP-activated protein kinase (AMPK) functions as a cellular energy gauge that maintains cellular homeostasis and has been suggested to play important roles in tumorigenesis, lifespan and autophagy. Accordingly, AMPK is a potential target of drugs for controlling a growing number of human diseases ranging from metabolic disorders to cancer, highlighting the need for rational and robust screening systems for identifying compounds that modulate AMPK.

Areas covered: The relevant screening methods in the patent and scientific literature were analyzed, and key features of direct AMPK modulators are discussed in the context of their physiological relevance and the three-dimensional structure of the AMPK complex.

Expert opinion: The mechanism of action of modulators is important in designing drugs with enhanced efficacy, specificity and stability. Most patented assay formats for identifying AMPK modulators are based on classical enzyme assays that monitor AMPK activity or changes in AMPK-dependent cellular physiology. However, these systems do not provide information about underlying mechanisms. Two patented assay systems use a specific domain or the three-dimensional structure of AMPK to identify AMPK modulators. The recent identification of two AMPK modulators, A-769662 and C-2 (or its prodrug, C-13), suggests the promise of structure-based assays in discovering more potent and specific modulators of AMPK.     

Exploiting the anti-inflammatory effects of AMP-activated protein kinase activation

Introduction: AMP-activated protein kinase (AMPK) is the downstream component of a serine/threonine protein kinase cascade involved in the regulation of metabolism. Many studies have also revealed that AMPK activation can exert significant anti-inflammatory and immunosuppressive effects in a variety of cell types and models of inflammatory/autoimmune disease. Because metformin, an AMPK activator that is a favored first-line therapeutic option for type 2 diabetes, may confer benefits in chronic inflammatory diseases and cancers independent of its ability to normalize blood glucose, there is now considerable interest in identifying and exploiting AMPK's anti-inflammatory effects.

Areas covered: The authors provide a background to AMPK signaling and describe the pro-inflammatory signaling pathways and processes shown to be regulated by AMPK activation.

Expert opinion: Identification of AMPK subunits responsible for specific anti-inflammatory effects, and a molecular understanding of the mechanisms involved, will be necessary to exploit AMPK pathway activation in acute and chronic inflammatory disease settings while minimizing adverse reactions due to deregulation of AMPK's wide-ranging effects on metabolism.     

WT-1蛋白在卵巢颗粒细胞瘤和卵泡膜细胞瘤中的表达及意义

目的探讨WT-1蛋白在卵巢颗粒细胞瘤和卵泡膜细胞瘤中的表达及意义。方法用免疫组织化学方法研究WT-1蛋白在37例卵巢颗粒细胞瘤(GCT)和39例卵泡膜细胞瘤(TCT)及10份健康卵巢组织中的表达。结果在卵巢GCT和TCT组织中,WT-1蛋白的表达明显高于其在正常组织中的表达。且高、中分化组与低分化组之间,Ⅰ期、Ⅱ期与Ⅲ期之间的差异均有统计学意义(P<0.05),即随组织学分级与临床分期提高而降低。WT-1蛋白阳性表达的病例10年生存率高于阴性表达者,差异有统计学意义(P<0.05)。结论WT-1蛋白有望作为卵巢GCT和TCT的辅助标记物和影响预后的一个因素,能为病理诊断和判定预后提供一定的指导作用。     

A comprehensive insight into potential roles of Nigella sativa on diseases by targeting AMP-activated protein kinase: a review

ObjectivesNigella sativa (NS) is a known medicinal herb with numerous therapeutic effects such as antidiabetic, anti-proliferative, anti-inflammatory, and anti-cancer activities. It has been indicated that NS can regulate cellular metabolism by adjusting transduction signaling pathways. Adenosine monophosphate-activated protein kinase (AMPK) is one of the main physiological processes, such as energy hemostasis, cellular metabolism, and autophagy regulators. Herb-derived medicines have always been considered as one of the main AMPK activators, and surprisingly recent data has demonstrated that it can be a target for NS and its derivatives.Evidence acquisitionThe literature search was conducted in PubMed, SCOPUS, Embase, ProQuest, and Google Scholar electronic resources. Published articles up to September 2020 were considered, and those of which investigated Nigella sativa effects on the AMPK pathway after meeting the inclusion criteria were included.ResultsThe search was performed on several online databases such as PubMed, Scopus, Embase, ProQuest, and Google Scholar from inception until January 2020. Among the initial search, 245 studies were found. After removing duplicated data and meeting the inclusion criteria, only 14 studies were selected. They included the effects of NS and its bioactive compounds as anti-hyperglycemic (n = 5), on liver function (n = 4), cancers (n = 3), and on Neuroinflammation and Atherosclerosis (n = 2). Most of the included studies are animals or in-vitro investigations.ConclusionIn this review, we discuss the latest findings on the molecular mechanism of NS effecting the AMPK signaling pathway. We also focus on the therapeutic effects of NS, including the prevention and treatment of metabolic and pro-inflammatory disease by targeting the AMPK pathway.Graphical abstract     

Salidroside stimulated glucose uptake in skeletal muscle cells by activating AMP-activated protein kinase

In the present study, we reported the metabolic effects of salidroside, one of the active components of Rhodiola Rosea, on skeletal muscle cells. Salidroside dose-dependently stimulated glucose uptake in differentiated L6 rat myoblast cells. Inhibitor of AMP-activated protein kinase (AMPK) by pretreating the cells with compound C potently reduced salidroside-stimulated glucose uptake, while inhibition of phosphatidylinositol 3-kinase (PI3K) by wortmannin exhibited no significant inhibitory effect on salidroside-mediated glucose transport activation. Western blotting analyses revealed that salidroside increased the phosphorylation level of AMPK and acetyl-CoA carboxylase (ACC). In addition, salidroside enhanced insulin-mediated Akt activation and glucose uptake, and such enhancement can be specifically inhibited by compound C. In summary, AMPK activation was involved in the effects of salidroside on glucose transport activation and insulin sensitivity. Salidroside can be further developed as potential compound for the anti-diabetic therapy.     

Pokemon蛋白和p53蛋白在卵巢癌组织中的表达及其意义

目的探讨卵巢癌组织中Pokemon蛋白和p53蛋白表达,明确其与临床病理的关系。方法选取52例卵巢癌组织作为观察组,60例卵巢良性病变组织作为对照组。免疫组化测定两组组织中Pokemon蛋白和p53蛋白的表达。结果观察组和对照组中Pokemon蛋白阳性率分别为69．2％（36／52）和11．7％（7／60）,差异有统计学意义（χ2=12．74,P〈0．01）。观察组和对照组中p53蛋白阳性率分别为78．8％（41／52）和15．0％（9／60）,差异有统计学意义（χ2=22．71,P〈0．01）。Pokemon蛋白表达与卵巢癌患者的肿瘤浸润深度（χ2=9．16,P〈0．05）、临床分期（χ2=5．86,P〈0．05）、淋巴结是否转移（χ2=32．28,P〈0．01）和是否复发（χ2=27．08,P〈0．01）均有显著的关系,与肿瘤分化程度（χ2=0．82,P〉0．05）无明显关系。p53蛋白表达与卵巢癌患者的肿瘤浸润深度（χ2=8．24,P〈0．05）、临床分期（χ2=9．24,P〈0．05）、淋巴结是否转移（χ2=19．62,P〈0．01）和是否复发（χ2=21．16,P〈0．01）均有显著的关系,与肿瘤分化程度（χ2=0．77,P〉0．05）无明显关系。经相关性分析,Pokemon蛋白和p53蛋白阳性表达有明显正相关关系（r=0．297,P〈0．05）。结论Pokemon蛋白和p53蛋白与卵巢癌的诊断、病变程度和预后均有密切关系。     

Activation of AMP-activated protein kinase by tributyltin induces neuronal cell death

AMP-activated protein kinase (AMPK), a member of the metabolite-sensing protein kinase family, is activated by energy deficiency and is abundantly expressed in neurons. The environmental pollutant, tributyltin chloride (TBT), is a neurotoxin, and has been reported to decrease cellular ATP in some types of cells. Therefore, we investigated whether TBT activates AMPK, and whether its activation contributes to neuronal cell death, using primary cultures of cortical neurons. Cellular ATP levels were decreased 0.5 h after exposure to 500 nM TBT, and the reduction was time-dependent. It was confirmed that most neurons in our culture system express AMPK, and that TBT induced phosphorylation of AMPK. Compound C, an AMPK inhibitor, reduced the neurotoxicity of TBT, suggesting that AMPK is involved in TBT-induced cell death. Next, the downstream target of AMPK activation was investigated. Nitric oxide synthase, p38 phosphorylation and Akt dephosphorylation were not downstream of TBT-induced AMPK activation because these factors were not affected by compound C, but glutamate release was suggested to be controlled by AMPK. Our results suggest that activation of AMPK by TBT causes neuronal death through mediating glutamate release.     

Perivascular fat,AMP-activated protein kinase and vascular diseases

Perivascular adipose tissue (PVAT) is an active endocrine and paracrine organ that modulates vascular function, with implications for the pathophysiology of cardiovascular disease (CVD). Adipocytes and stromal cells contained within PVAT produce mediators (adipokines, cytokines, reactive oxygen species and gaseous compounds) with a range of paracrine effects modulating vascular smooth muscle cell contraction, proliferation and migration. However, the modulatory effect of PVAT on the vascular system in diseases, such as obesity, hypertension and atherosclerosis, remains poorly characterized. AMP-activated protein kinase (AMPK) regulates adipocyte metabolism, adipose biology and vascular function, and hence may be a potential therapeutic target for metabolic disorders such as type 2 diabetes mellitus (T2DM) and the vascular complications associated with obesity and T2DM. The role of AMPK in PVAT or the actions of PVAT have yet to be established, however. Activation of AMPK by pharmacological agents, such as metformin and thiazolidinediones, may modulate the activity of PVAT surrounding blood vessels and thereby contribute to their beneficial effect in cardiometabolic diseases. This review will provide a current perspective on how PVAT may influence vascular function via AMPK. We will also attempt to demonstrate how modulating AMPK activity using pharmacological agents could be exploited therapeutically to treat cardiometabolic diseases.     

Direct AMP-activated protein kinase activators: a review of evidence from the patent literature

Introduction: AMP-activated protein kinase (AMPK), a heterotrimeric protein complex with serine/threonine kinase activity has a central role in controlling cellular energy expenditure. Small molecule-based activation of AMPK represents an attractive therapeutic proposition because of AMPK's ability to regulate several anabolic and catabolic pathways that are critical to the development of metabolic disorders and cancer.

Areas covered: A comprehensive review of published patents that disclose direct AMPK activators is provided: 26 patents comprising 10 chemical classes, and supporting in vitro and in vivo data are discussed.

Expert opinion: AMPK activation holds promise as a possible pharmacological intervention in several disease states. The development of direct, highly specific AMPK activators is necessary to fully realize the opportunities linked to AMPK activation and appreciate the risks associated with it.     

Metformin-induced stimulation of AMP-activated protein kinase in beta-cells impairs their glucose responsiveness and can lead to apoptosis

Metformin is an anti-diabetic drug that increases glucose utilization in insulin-sensitive tissues. The effect is in part attributable to a stimulation of AMP-activated protein kinase (AMPK). The present study demonstrates that metformin (0.5-2mM) also dose-dependently activates AMPK in insulin-producing MIN6 cells and in primary rat beta-cells, leading to increased phosphorylation of acetyl coA carboxylase (ACC). The maximal effect was reached within 12h and sustained up to 48h. After 24h exposure to metformin (0.5-1mM), rat beta-cells exhibited a reduced secretory and synthetic responsiveness to 10mM glucose, which was also the case following 24h culture with the AMPK-activator 5-amino-imidazole-4-carboxamide riboside (AICAR; 1mM). Longer metformin exposure (>24h) resulted in a progressive increase in apoptotic beta-cells as was also reported for AICAR; metformin-induced apoptosis was reduced by compound C, an AMPK-inhibitor. As with AICAR, metformin activated c-Jun-N-terminal kinase (JNK) and caspase-3 prior to the appearance of apoptosis. It is concluded that metformin-induced AMPK-activation in beta-cells reduces their glucose responsiveness and may, following sustained exposure, result in apoptosis.