The incidence and mortality rate of lung cancer rank among the highest worldwide,severely endangering human health and life.Metformin,an anti-diabetes drug,has been shown to elicit anticancer activities in various tum...The incidence and mortality rate of lung cancer rank among the highest worldwide,severely endangering human health and life.Metformin,an anti-diabetes drug,has been shown to elicit anticancer activities in various tumors.However,its underlying mechanisms remain elusive.In this work,we explore the role of receptor-interacting protein 1(RIP1)which plays a crucial role in the process of cell death,in metformin-induced anticancer activities in lung cancer.Metformin inhibits lung cancer cell proliferation in a dose-dependent manner and promotes apoptotic cell death,as evidenced by metformin-induced PARP and caspase cleavage.Furthermore,the pan-caspase inhibitor z-VAD-fmk reverses metformin-induced cell death.Western blot and qPCR results suggest that metformin markedly downregulates RIP1 expression without affecting its mRNA and ubiquitination levels(0 vs 80 mmol/L,100%vs 20%,100%vs 15%).Additionally,co-immunoprecipitation and immunofluorescence results reveal that metformin may suppress RIP1 expression in an Hsp70-dependent manner,as metformin promotes Hsp70 degradation,and Hsp70 endogenously interacts with RIP1.Subsequent CCK-8,flow cytometry,and Western blot analyses suggest that metformin decreases Hsp70/RIP1 expression through AMPK/PKA/GSK-3βaxis.Consistently,results from a subcutaneous transplant tumor model indicate that metformin retards tumor growth without affecting mouse body weight.Collectively,these data highlight the part of RIP1 in metformin-induced anticancer activities in lung cancer in vitro and in vivo,providing novel strategy for lung cancer administration.展开更多
AIM:Regulatory T cells(Tregs)are a specialized subset of CD4^(+)T cells primarily involved in im⁃munosuppressive functions.AMP-activated protein kinase(AMPK)serves as a metabolic sensor that governs the differen⁃tiati...AIM:Regulatory T cells(Tregs)are a specialized subset of CD4^(+)T cells primarily involved in im⁃munosuppressive functions.AMP-activated protein kinase(AMPK)serves as a metabolic sensor that governs the differen⁃tiation,maturation,and immune functions of Tregs through metabolic reprogramming.However,the impact of AMPKα1(the catalytic subunit of AMPK)knockout specifically in Tregs on the host's immune microenvironment remains largely un⁃explored.METHODS:Histological changes in immune organs were assessed using HE staining.The types of immune cells and their relative population percentages in immune organs and blood were quantified through flow cytometry in both AMPKα1flox/flox(AMPKα1^(fl/fl))mice and Treg-specific AMPKα1 knockout mice(AMPKα1^(fl/fl)Foxp3^(cre)mice).RESULTS:Compared to AMPKα1^(fl/fl)mice,the percentage of eosinophils in the bone marrow of AMPKα1^(fl/fl)Foxp3^(cre)mice was significant⁃ly reduced.Additionally,while the thymus of AMPKα1^(fl/fl)Foxp3^(cre)mice exhibited normal structure,both its size and the ratio of thymus weight to body weight were significantly decreased.The knockout of AMPKα1 in Tregs led to a notable reduction in the total percentage of immature double-negative(DN)cells.Consequently,the percentage of CD4^(+)T cells derived from these DN cells also decreased,even though the percentages of DN1 and DN4 cells were higher in the thymus of AMPKα1^(fl/fl)Foxp3^(cre)mice compared to AMPKα1^(fl/fl)mice.Importantly,the proportion of Siglec-F+CD11b^(+)eosinophils in the thymus was significantly lower in AMPKα1^(fl/fl)Foxp3^(cre)mice.Knockout of AMPKα1 in Tregs resulted in a marked increase in the percentage of CD4^(+)T cells in peripheral blood,alongside a decrease in the proportion of mature CD8^(+)T cells.Similarly,the proportion of CD4^(+)T cells in the spleen of AMPKα1^(fl/fl)Foxp3^(cre)mice was elevated compared to AMPKα1^(fl/fl)mice.In contrast,the proportion of neutrophils significantly decreased,while mononuclear cell proportions increased in the spleen of AMPKα1^(fl/fl)Foxp3^(cre)mice.In lymph nodes,the medullary boundaries in AMPKα1^(fl/fl)Foxp3^(cre)mice were blurred,and the lymphoid follicles were missing,a feature not observed in AMPKα1^(fl/fl)mice.Furthermore,the knockout of AMPKα1 in Tregs reduced the CD3^(+)T cell population,particularly the CD8^(+)T cell population,in lymph nodes.Although the mature Treg cell population was significantly lower in AMPKα1^(fl/fl)Foxp3^(cre)mice,the percentage of CD4^(+)T cells was markedly in⁃creased.In contrast,there was no statistically significant difference in granulocyte populations between AMPKα1^(fl/fl)Foxp3^(cre)and AMPKα1^(fl/fl)mice.CONCLUSION:The populations of mature Tregs,CD8^(+)T cells and eosinophils in various im⁃mune organs were significantly altered in mice with Treg-specific AMPKα1 knockout,suggesting a potential remodeling of the host immune microenvironment in response to inflammatory stimuli.展开更多
多囊卵巢综合征(Polycystic ovary syndrome,PCOS)是一组生殖内分泌代谢紊乱的综合征,临床以稀发排卵、高雄激素体征、胰岛素抵抗为主要特征,其中育龄期发病率高,对女性生育力造成严重不良影响。PCOS的发生发展涉及多种信号通路,腺苷酸...多囊卵巢综合征(Polycystic ovary syndrome,PCOS)是一组生殖内分泌代谢紊乱的综合征,临床以稀发排卵、高雄激素体征、胰岛素抵抗为主要特征,其中育龄期发病率高,对女性生育力造成严重不良影响。PCOS的发生发展涉及多种信号通路,腺苷酸活化蛋白激酶(AMP-activated protein kinase,AMPK)及哺乳动物雷帕霉素靶蛋白(Mammalian target of rapamycin,mTOR)作为细胞能量感受器是其中两个关键靶点。二者在PCOS各个发病部位包括下丘脑-垂体-卵巢轴、子宫内膜、脂肪与骨骼肌中发挥重要的调节作用,通过影响细胞自噬、氧化应激、炎症、线粒体功能、葡萄糖摄取等,促进卵泡的发育和成熟,改善胰岛素抵抗。近年来,中医药因其成分多样、靶点众多等优势广泛应用于临床,研究人员已对PCOS的发病以及中药治疗及改善PCOS的机制进行了大量研究,结果提示AMPK与mTOR相关通路在其中发挥关键作用。通过总结中药干预AMPK与mTOR及其相关通路治疗PCOS的研究结果,为临床治疗及基础研究提供参考。展开更多
目的:研究祛湿化瘀方对高脂饮食诱导的大鼠脂肪肝AMPK蛋白活性及其相关脂肪代谢靶蛋白活性的影响,以探讨该方防治实验性脂肪肝的作用机制。方法:采用高脂饲料饮食诱导大鼠脂肪肝模型,造模大鼠给予高脂饮食4周后,按随机数字表随机分为模...目的:研究祛湿化瘀方对高脂饮食诱导的大鼠脂肪肝AMPK蛋白活性及其相关脂肪代谢靶蛋白活性的影响,以探讨该方防治实验性脂肪肝的作用机制。方法:采用高脂饲料饮食诱导大鼠脂肪肝模型,造模大鼠给予高脂饮食4周后,按随机数字表随机分为模型组及祛湿化瘀方组,分别灌胃给予饮用水及中药祛湿化瘀方4周。实验8周末取材后观察:1)肝组织三酰甘油(Triglyceride,TG)、游离脂肪酸(Free Fatty Acid,FFA)含量,2)肝组织病理变化(HE染色、油红染色),3)肝组织腺苷酸活化的蛋白激酶(AMP-Activated K inase,AMPK)及磷酸化AMPK、肝组织总蛋白及核蛋白固醇调节元件结合蛋白-1c(Sterol Regulatory Element Binding Protein-1c、SREBP-1c)、肝组织总蛋白及核蛋白碳水化合物反应元件结合蛋白(Carbohydrate Response Element Binding Protein、ChREBP)含量、肝组织乙酰辅酶A羧化酶(Acety1 Co A Carboxylase,ACCase)及磷酸化ACC蛋白含量,4)肝组织AMPKα1、AMPKα2、SREBP-1、ACCα、SREBP-1c及ChREBP基因表达水平。结果:1)模型组肝组织TG、FFA含量显著升高,肝组织出现明显大泡样脂肪变性;模型组肝组织AMPK蛋白磷酸化水平降低、核蛋白SREBP-1与ChREBP表达增加、ACC蛋白磷酸化水平降低蛋白活性升高。2)祛湿化瘀方组肝组织TG、FFA含量较模型组显著降低,肝组织炎症及脂肪变性程度减轻;祛湿化瘀方能显著升高肝组织AMPK、ACC蛋白磷酸化水平、降低核蛋白SREBP-1及ChREBP含量。结论:祛湿化瘀方通过调节AMPK活性及其相关靶蛋白活性改善高脂饮食诱导的大鼠脂肪肝肝脂肪代谢,这可能是该方有效防治实验性脂肪肝的重要作用机制之一。展开更多
二甲双胍是一种传统的口服降糖药,临床上普遍用于2型糖尿病的治疗。近年来大量流行病学研究报道二甲双胍能够降低2型糖尿病患者的肿瘤发病率,亦有研究发现二甲双胍能在代谢途径、细胞周期、氧化应激、肿瘤干细胞转化等方面通过激活腺苷...二甲双胍是一种传统的口服降糖药,临床上普遍用于2型糖尿病的治疗。近年来大量流行病学研究报道二甲双胍能够降低2型糖尿病患者的肿瘤发病率,亦有研究发现二甲双胍能在代谢途径、细胞周期、氧化应激、肿瘤干细胞转化等方面通过激活腺苷酸活化蛋白激酶(adenosin emonophosphate-activated protein kinase,AMPK)信号通路,从而抑制肿瘤细胞的生长、增殖以及转化。但二甲双胍通过激活AMPK的抗肿瘤机制仍存在着争议,其确切的作用机制有待进一步深入的研究,同时亟需大规模的临床试验来证实。展开更多
2型糖尿病是一种慢性代谢性疾病,其主要表现为高血糖和微血管并发症,是影响人类生活的重大疾病之一,目前尚无理想的治疗药物。腺苷酸活化蛋白激酶(AMP-activated protein kinase,AMPK)是一种高度保守的丝氨酸/苏氨酸蛋白激酶,在细胞和...2型糖尿病是一种慢性代谢性疾病,其主要表现为高血糖和微血管并发症,是影响人类生活的重大疾病之一,目前尚无理想的治疗药物。腺苷酸活化蛋白激酶(AMP-activated protein kinase,AMPK)是一种高度保守的丝氨酸/苏氨酸蛋白激酶,在细胞和机体能量代谢与平衡等方面发挥重要作用;小分子AMPK直接激动剂因在降低血糖方面的作用而有望成为新一代治疗2型糖尿病的药物。本文按结构分类,对近几年报道的小分子AMPK直接激动剂进行综述。展开更多
文摘The incidence and mortality rate of lung cancer rank among the highest worldwide,severely endangering human health and life.Metformin,an anti-diabetes drug,has been shown to elicit anticancer activities in various tumors.However,its underlying mechanisms remain elusive.In this work,we explore the role of receptor-interacting protein 1(RIP1)which plays a crucial role in the process of cell death,in metformin-induced anticancer activities in lung cancer.Metformin inhibits lung cancer cell proliferation in a dose-dependent manner and promotes apoptotic cell death,as evidenced by metformin-induced PARP and caspase cleavage.Furthermore,the pan-caspase inhibitor z-VAD-fmk reverses metformin-induced cell death.Western blot and qPCR results suggest that metformin markedly downregulates RIP1 expression without affecting its mRNA and ubiquitination levels(0 vs 80 mmol/L,100%vs 20%,100%vs 15%).Additionally,co-immunoprecipitation and immunofluorescence results reveal that metformin may suppress RIP1 expression in an Hsp70-dependent manner,as metformin promotes Hsp70 degradation,and Hsp70 endogenously interacts with RIP1.Subsequent CCK-8,flow cytometry,and Western blot analyses suggest that metformin decreases Hsp70/RIP1 expression through AMPK/PKA/GSK-3βaxis.Consistently,results from a subcutaneous transplant tumor model indicate that metformin retards tumor growth without affecting mouse body weight.Collectively,these data highlight the part of RIP1 in metformin-induced anticancer activities in lung cancer in vitro and in vivo,providing novel strategy for lung cancer administration.
基金Supported by the National Natural Science Foundation of China(No.81800423)the Guangdong Medical Science and Technology Research project(No.B2022102)。
文摘AIM:Regulatory T cells(Tregs)are a specialized subset of CD4^(+)T cells primarily involved in im⁃munosuppressive functions.AMP-activated protein kinase(AMPK)serves as a metabolic sensor that governs the differen⁃tiation,maturation,and immune functions of Tregs through metabolic reprogramming.However,the impact of AMPKα1(the catalytic subunit of AMPK)knockout specifically in Tregs on the host's immune microenvironment remains largely un⁃explored.METHODS:Histological changes in immune organs were assessed using HE staining.The types of immune cells and their relative population percentages in immune organs and blood were quantified through flow cytometry in both AMPKα1flox/flox(AMPKα1^(fl/fl))mice and Treg-specific AMPKα1 knockout mice(AMPKα1^(fl/fl)Foxp3^(cre)mice).RESULTS:Compared to AMPKα1^(fl/fl)mice,the percentage of eosinophils in the bone marrow of AMPKα1^(fl/fl)Foxp3^(cre)mice was significant⁃ly reduced.Additionally,while the thymus of AMPKα1^(fl/fl)Foxp3^(cre)mice exhibited normal structure,both its size and the ratio of thymus weight to body weight were significantly decreased.The knockout of AMPKα1 in Tregs led to a notable reduction in the total percentage of immature double-negative(DN)cells.Consequently,the percentage of CD4^(+)T cells derived from these DN cells also decreased,even though the percentages of DN1 and DN4 cells were higher in the thymus of AMPKα1^(fl/fl)Foxp3^(cre)mice compared to AMPKα1^(fl/fl)mice.Importantly,the proportion of Siglec-F+CD11b^(+)eosinophils in the thymus was significantly lower in AMPKα1^(fl/fl)Foxp3^(cre)mice.Knockout of AMPKα1 in Tregs resulted in a marked increase in the percentage of CD4^(+)T cells in peripheral blood,alongside a decrease in the proportion of mature CD8^(+)T cells.Similarly,the proportion of CD4^(+)T cells in the spleen of AMPKα1^(fl/fl)Foxp3^(cre)mice was elevated compared to AMPKα1^(fl/fl)mice.In contrast,the proportion of neutrophils significantly decreased,while mononuclear cell proportions increased in the spleen of AMPKα1^(fl/fl)Foxp3^(cre)mice.In lymph nodes,the medullary boundaries in AMPKα1^(fl/fl)Foxp3^(cre)mice were blurred,and the lymphoid follicles were missing,a feature not observed in AMPKα1^(fl/fl)mice.Furthermore,the knockout of AMPKα1 in Tregs reduced the CD3^(+)T cell population,particularly the CD8^(+)T cell population,in lymph nodes.Although the mature Treg cell population was significantly lower in AMPKα1^(fl/fl)Foxp3^(cre)mice,the percentage of CD4^(+)T cells was markedly in⁃creased.In contrast,there was no statistically significant difference in granulocyte populations between AMPKα1^(fl/fl)Foxp3^(cre)and AMPKα1^(fl/fl)mice.CONCLUSION:The populations of mature Tregs,CD8^(+)T cells and eosinophils in various im⁃mune organs were significantly altered in mice with Treg-specific AMPKα1 knockout,suggesting a potential remodeling of the host immune microenvironment in response to inflammatory stimuli.
文摘多囊卵巢综合征(Polycystic ovary syndrome,PCOS)是一组生殖内分泌代谢紊乱的综合征,临床以稀发排卵、高雄激素体征、胰岛素抵抗为主要特征,其中育龄期发病率高,对女性生育力造成严重不良影响。PCOS的发生发展涉及多种信号通路,腺苷酸活化蛋白激酶(AMP-activated protein kinase,AMPK)及哺乳动物雷帕霉素靶蛋白(Mammalian target of rapamycin,mTOR)作为细胞能量感受器是其中两个关键靶点。二者在PCOS各个发病部位包括下丘脑-垂体-卵巢轴、子宫内膜、脂肪与骨骼肌中发挥重要的调节作用,通过影响细胞自噬、氧化应激、炎症、线粒体功能、葡萄糖摄取等,促进卵泡的发育和成熟,改善胰岛素抵抗。近年来,中医药因其成分多样、靶点众多等优势广泛应用于临床,研究人员已对PCOS的发病以及中药治疗及改善PCOS的机制进行了大量研究,结果提示AMPK与mTOR相关通路在其中发挥关键作用。通过总结中药干预AMPK与mTOR及其相关通路治疗PCOS的研究结果,为临床治疗及基础研究提供参考。
文摘目的:研究祛湿化瘀方对高脂饮食诱导的大鼠脂肪肝AMPK蛋白活性及其相关脂肪代谢靶蛋白活性的影响,以探讨该方防治实验性脂肪肝的作用机制。方法:采用高脂饲料饮食诱导大鼠脂肪肝模型,造模大鼠给予高脂饮食4周后,按随机数字表随机分为模型组及祛湿化瘀方组,分别灌胃给予饮用水及中药祛湿化瘀方4周。实验8周末取材后观察:1)肝组织三酰甘油(Triglyceride,TG)、游离脂肪酸(Free Fatty Acid,FFA)含量,2)肝组织病理变化(HE染色、油红染色),3)肝组织腺苷酸活化的蛋白激酶(AMP-Activated K inase,AMPK)及磷酸化AMPK、肝组织总蛋白及核蛋白固醇调节元件结合蛋白-1c(Sterol Regulatory Element Binding Protein-1c、SREBP-1c)、肝组织总蛋白及核蛋白碳水化合物反应元件结合蛋白(Carbohydrate Response Element Binding Protein、ChREBP)含量、肝组织乙酰辅酶A羧化酶(Acety1 Co A Carboxylase,ACCase)及磷酸化ACC蛋白含量,4)肝组织AMPKα1、AMPKα2、SREBP-1、ACCα、SREBP-1c及ChREBP基因表达水平。结果:1)模型组肝组织TG、FFA含量显著升高,肝组织出现明显大泡样脂肪变性;模型组肝组织AMPK蛋白磷酸化水平降低、核蛋白SREBP-1与ChREBP表达增加、ACC蛋白磷酸化水平降低蛋白活性升高。2)祛湿化瘀方组肝组织TG、FFA含量较模型组显著降低,肝组织炎症及脂肪变性程度减轻;祛湿化瘀方能显著升高肝组织AMPK、ACC蛋白磷酸化水平、降低核蛋白SREBP-1及ChREBP含量。结论:祛湿化瘀方通过调节AMPK活性及其相关靶蛋白活性改善高脂饮食诱导的大鼠脂肪肝肝脂肪代谢,这可能是该方有效防治实验性脂肪肝的重要作用机制之一。
文摘二甲双胍是一种传统的口服降糖药,临床上普遍用于2型糖尿病的治疗。近年来大量流行病学研究报道二甲双胍能够降低2型糖尿病患者的肿瘤发病率,亦有研究发现二甲双胍能在代谢途径、细胞周期、氧化应激、肿瘤干细胞转化等方面通过激活腺苷酸活化蛋白激酶(adenosin emonophosphate-activated protein kinase,AMPK)信号通路,从而抑制肿瘤细胞的生长、增殖以及转化。但二甲双胍通过激活AMPK的抗肿瘤机制仍存在着争议,其确切的作用机制有待进一步深入的研究,同时亟需大规模的临床试验来证实。
文摘2型糖尿病是一种慢性代谢性疾病,其主要表现为高血糖和微血管并发症,是影响人类生活的重大疾病之一,目前尚无理想的治疗药物。腺苷酸活化蛋白激酶(AMP-activated protein kinase,AMPK)是一种高度保守的丝氨酸/苏氨酸蛋白激酶,在细胞和机体能量代谢与平衡等方面发挥重要作用;小分子AMPK直接激动剂因在降低血糖方面的作用而有望成为新一代治疗2型糖尿病的药物。本文按结构分类,对近几年报道的小分子AMPK直接激动剂进行综述。
