目的:探讨有氧运动对不同年龄小鼠海马哺乳动物雷帕霉素靶蛋白(mammalian target of rapamycin,mTOR)信号通路活性的影响及其与认知功能的关系。方法:将1月龄C57BL/6雄性小鼠90只,随机分为有氧运动(E)组(n=45)和安静对照(C)组(n=45),有...目的:探讨有氧运动对不同年龄小鼠海马哺乳动物雷帕霉素靶蛋白(mammalian target of rapamycin,mTOR)信号通路活性的影响及其与认知功能的关系。方法:将1月龄C57BL/6雄性小鼠90只,随机分为有氧运动(E)组(n=45)和安静对照(C)组(n=45),有氧运动组小鼠进行75%VO2max强度的有氧跑台运动干预;两组分别至小鼠3、12、20月龄时,行Morris水迷宫实验测试认知功能后,处死取材(n=15);采用Western blot法检测小鼠海马mTOR信号通路蛋白mTOR、mTOR丝氨酸2448位点磷酸化[p-mTOR(Ser2448)]、核糖体S6蛋白激酶1苏氨酸389位点磷酸化[p-S6K1(Thr389)]、真核生物翻译起始因子4E结合蛋白丝氨酸65位点磷酸化[p-4EBP1(Ser65)]及其上游脑源性神经营养因子(brainderived neurotrophic factor,BDNF)表达水平;采用免疫组织化学染色法检测小鼠海马BDNF蛋白阳性细胞并计算积分光密度。结果:水迷宫定位航行试验第3天开始,12月龄和20月龄小鼠逃避潜伏期显著长于3月龄小鼠(P<0.05,P<0.01),且20月龄较12月龄逃避潜伏期更长(P<0.05,P<0.01);目标象限航程和原平台位置穿越次数,12月龄小鼠显著低于3月龄小鼠(P<0.05),20月龄小鼠目标象限航程显著低于3、12月龄小鼠(P<0.01)。与同龄对照组相比,运动组逃避潜伏期显著缩短(P<0.05),目标象限航程显著增加(P<0.01)。20月龄小鼠p-mTOR(Ser2448)、p-S6K1(Thr389)、p-4EBP1(Ser65)及BDNF表达水平显著低于3月龄或12月龄小鼠(P<0.05,P<0.01);对于3月龄小鼠海马p-mTOR(Ser2448)、p-S6K1(Thr389)、p-4EBP1(Ser65)表达水平,运动组显著高于对照组(P<0.01);对于12和20月龄小鼠p-mTOR(Ser2448)、p-S6K1(Thr389)、p-4EBP1(Ser65)表达水平,运动组显著低于同龄对照组(P<0.05,P<0.01);与同龄对照组相比,运动组小鼠海马BDNF的表达水平显著提高(P<0.01)。结论:有氧运动促进幼年小鼠认知功能发育并延缓老年小鼠认知功能增龄性退变,这一作用效果可能通过激活幼年小鼠海马mTOR信号通路或抑制成年后该信号通路活性而实现;且提示mTOR信号通路活性对发育期小鼠海马记忆的必要性,但其过度活跃对老年小鼠认知可能产生不利影响。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
Objective:IgA nephropathy(IgAN)is the most common primary glomerular disease in China,but its pathogenesis remains unclear.This study aims to explore the regulatory role of the mammalian target of rapamycin(mTOR)signa...Objective:IgA nephropathy(IgAN)is the most common primary glomerular disease in China,but its pathogenesis remains unclear.This study aims to explore the regulatory role of the mammalian target of rapamycin(mTOR)signaling pathway in autophagy and mesangial proliferation during renal injury in IgA.Methods:The activity of mTOR and autophagy was evaluated in kidney samples from IgAN patients and in an IgAN mouse model induced by oral bovine serum albumin and carbon tetrachloride(CCl4)injection.mTOR inhibitors(rapamycin)and activators[bpV(phen)]were administered to the IgAN mouse model to observe the effects of mTOR on autophagy and renal lesions.In human mesangial cells treated with polymeric IgA1(p IgA1)and mTOR modulators,the expression and distribution of cell cycle proteins were assessed,along with the effects of mTOR on mesangial cell proliferation and autophagy.Results:Increased mTOR activity and decreased autophagy were observed in kidney tissues from IgAN patients and the mouse model,as evidenced by elevated phosphorylated mTOR(p-mTOR)levels and reduced LC3 expression.In the IgAN mouse model,rapamycin inhibited mTOR,restored autophagy,reduced mesangial IgA deposition,alleviated mesangial cell proliferation,and decreased proteinuria(all P<0.05).In contrast,bpV(phen)activated mTOR,further suppressed autophagy,exacerbated kidney damage,and increased proteinuria(all P<0.05).In vitro,p-IgA1 induced mesangial cell proliferation and inhibited autophagy,effects that were reversed by rapamycin and aggravated by bpV(phen)(all P<0.05).mTOR regulated mesangial cell proliferation by altering cell cycle distribution,with rapamycin inducing G1 phase arrest and bpV(phen)promoting cell cycle progression.Additionally,cyclinD1 expression in renal cortex was up-regulated in the IgAN mouse model,further increased by bpV(phen),and reduced by rapamycin(all P<0.05).Conclusion:Inhibition of the mTOR signaling pathway enhances renal autophagy,reduces mesangial cell proliferation,and improves renal injury in IgAN.展开更多
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。
基金supported by the Key Funding Project of Hunan Provincial Health Commission(A202303057091)the Nephrology Medical Development Research Fund Project(20220316ZN),China.
文摘Objective:IgA nephropathy(IgAN)is the most common primary glomerular disease in China,but its pathogenesis remains unclear.This study aims to explore the regulatory role of the mammalian target of rapamycin(mTOR)signaling pathway in autophagy and mesangial proliferation during renal injury in IgA.Methods:The activity of mTOR and autophagy was evaluated in kidney samples from IgAN patients and in an IgAN mouse model induced by oral bovine serum albumin and carbon tetrachloride(CCl4)injection.mTOR inhibitors(rapamycin)and activators[bpV(phen)]were administered to the IgAN mouse model to observe the effects of mTOR on autophagy and renal lesions.In human mesangial cells treated with polymeric IgA1(p IgA1)and mTOR modulators,the expression and distribution of cell cycle proteins were assessed,along with the effects of mTOR on mesangial cell proliferation and autophagy.Results:Increased mTOR activity and decreased autophagy were observed in kidney tissues from IgAN patients and the mouse model,as evidenced by elevated phosphorylated mTOR(p-mTOR)levels and reduced LC3 expression.In the IgAN mouse model,rapamycin inhibited mTOR,restored autophagy,reduced mesangial IgA deposition,alleviated mesangial cell proliferation,and decreased proteinuria(all P<0.05).In contrast,bpV(phen)activated mTOR,further suppressed autophagy,exacerbated kidney damage,and increased proteinuria(all P<0.05).In vitro,p-IgA1 induced mesangial cell proliferation and inhibited autophagy,effects that were reversed by rapamycin and aggravated by bpV(phen)(all P<0.05).mTOR regulated mesangial cell proliferation by altering cell cycle distribution,with rapamycin inducing G1 phase arrest and bpV(phen)promoting cell cycle progression.Additionally,cyclinD1 expression in renal cortex was up-regulated in the IgAN mouse model,further increased by bpV(phen),and reduced by rapamycin(all P<0.05).Conclusion:Inhibition of the mTOR signaling pathway enhances renal autophagy,reduces mesangial cell proliferation,and improves renal injury in IgAN.
