A recent study in the World Journal of Diabetes by Yang et al explored how Rheb1 signaling influenced pancreaticβcell fate and its potential as a therapeutic target.This invited commentary by a senior diabetes resear...A recent study in the World Journal of Diabetes by Yang et al explored how Rheb1 signaling influenced pancreaticβcell fate and its potential as a therapeutic target.This invited commentary by a senior diabetes researcher discussed the findings of Yang et al in the context of current knowledge onβcell biology,providing critical insight into the role of Rheb1 inβcell survival and function and the prospects for diabetes treatment.Key outcomes of the study were interpreted alongside established literature on Rheb1-mechanistic target of rapamycin signaling in islet cells.Rheb1 emerges as a pivotal regulator ofβcell growth and insulin secretory function,aligning with evidence thatβcell-specific Rheb1 deletion impairsβcell mass and glucose-stimulated insulin secretion.The commentary highlighted how modulating this pathway could preserve or restore theβcell population in diabetes while cautioning about potential off-target effects(e.g.inαcells).Targeting Rheb1 signaling represents a promising new frontier in diabetes therapy to enhanceβcell resilience;however,a balanced approach addressing both its benefits and risks is essential.This letter discussed the scientific implications and future research directions needed to translate Rheb1 modulation into clinical application for diabetes.展开更多
目的:利用前体脂肪细胞株3T3-L1细胞观察mTOR(mammalian target of rapamycin)信号通路中上游调控因子Rheb(Ras homolog enriched in brain)对其分化的影响。方法:利用高表达Rheb的基因重组质粒转染前体脂肪细胞株,3T3-L1。通过蛋白质...目的:利用前体脂肪细胞株3T3-L1细胞观察mTOR(mammalian target of rapamycin)信号通路中上游调控因子Rheb(Ras homolog enriched in brain)对其分化的影响。方法:利用高表达Rheb的基因重组质粒转染前体脂肪细胞株,3T3-L1。通过蛋白质免疫印迹实验鉴定质粒成功转染细胞后,诱导该细胞脂肪分化。予以分化第8天的3T3-L1细胞油红染色,并检测细胞内甘油三酯的含量。另外,我们用Western blot方法检测脂肪细胞特异性转录因子PPAR-γ(Peroxisome proliferator-activated receptor-γ)和C/EBP-α(CCAAT-enhancer-binding protein-α)的表达情况来研究Rheb在脂肪细胞分化过程中的作用。结果:我们成功构建了高表达Rheb的3T3-L1细胞株,发现高表达Rheb后可以促进脂滴的生成,油红O染色有显著区别,与对照组相比Rheb高表达组的三酰甘油含量明显升高(P<0.05);C/EBP-α和PPAR-γ等脂肪细胞特异性的转录因子蛋白表达量与对照组相比也均有升高(P<0.05)。结论:Rheb基因作为mTOR通路上游调控因子,可以促进脂肪细胞的分化。展开更多
【目的】研究旨在对努比亚山羊大脑富集Ras同源物(Ras homolog enriched in brain,Rheb)基因进行克隆和分析,并构建其真核表达载体,为进一步揭示Rheb对努比亚山羊骨骼肌调控的分子机理奠定基础。【方法】试验采用RT-PCR方法从努比亚山...【目的】研究旨在对努比亚山羊大脑富集Ras同源物(Ras homolog enriched in brain,Rheb)基因进行克隆和分析,并构建其真核表达载体,为进一步揭示Rheb对努比亚山羊骨骼肌调控的分子机理奠定基础。【方法】试验采用RT-PCR方法从努比亚山羊背最长肌组织中扩增Rheb基因,经琼脂糖凝胶电泳及测序验证正确后进行生物信息学分析,同时构建该基因真核表达载体,转染细胞后进行实时荧光定量PCR检测来验证所构建载体的正确性。【结果】努比亚山羊Rheb基因编码区序列长度为555 bp,编码184个氨基酸,Rheb蛋白分子式为C 910 H 1456 N 236 O 279 S 6,分子质量为20359.34 u,原子总数为2887,理论等电点为5.93。Rheb蛋白属于稳定的亲水性蛋白,不包含跨膜结构域。蛋白二级结构预测结果显示,努比亚山羊Rheb蛋白中α-螺旋、β-转角、延伸链和无规则卷曲分别占40.76%、7.07%、22.83%和29.35%。构建的真核表达载体pcDNA3.1-Rheb转染山羊骨骼肌细胞后,与空载体组相比,Rheb基因表达量极显著升高(P<0.01)。【结论】本试验成功克隆努比亚山羊Rheb基因编码区序列,并构建pcDNA3.1-Rheb真核表达载体,这为深入理解Rheb基因在努比亚山羊肌肉中的作用提供了理论支持。展开更多
Dysregulation of microRNAs(miRNAs)in adipocytes plays a critical role in the pathogenesis of obesity.However,the signaling mechanisms regulating miRNAs production in adipose tissue remain largely unclear.Here,we show ...Dysregulation of microRNAs(miRNAs)in adipocytes plays a critical role in the pathogenesis of obesity.However,the signaling mechanisms regulating miRNAs production in adipose tissue remain largely unclear.Here,we show that adipose tissue-specific knockout of Ras homolog enriched in brain(Rheb),a direct upstream activator of mTOR,increases miR-182-5p level in mouse subcutaneous white adipose tissues.Interestingly,the inhibition of mTOR signaling by rapamycin has no effect on miR-182-5p level in primary subcutaneous white adipocytes,suggesting the presence of a mTOR-independent mechanism regulating Rheb-mediated miR-182-5p expression.Consistent with this view,Rheb-ablation activates the cAMP/PPARγsignaling pathway.In addition,treatment of white adipocytes with pioglitazone,a PPARγagonist,dramatically upregulates miR-182-5p levels.Our study reveals a unique mechanism by which Rheb regulates miR-182-5p in adipocytes.Given that increasing miR-182-5p in adipose tissue promotes beige fat development,our study also suggests a unique mechanism by which Rheb promotes thermogenesis and energy expenditure.展开更多
文摘A recent study in the World Journal of Diabetes by Yang et al explored how Rheb1 signaling influenced pancreaticβcell fate and its potential as a therapeutic target.This invited commentary by a senior diabetes researcher discussed the findings of Yang et al in the context of current knowledge onβcell biology,providing critical insight into the role of Rheb1 inβcell survival and function and the prospects for diabetes treatment.Key outcomes of the study were interpreted alongside established literature on Rheb1-mechanistic target of rapamycin signaling in islet cells.Rheb1 emerges as a pivotal regulator ofβcell growth and insulin secretory function,aligning with evidence thatβcell-specific Rheb1 deletion impairsβcell mass and glucose-stimulated insulin secretion.The commentary highlighted how modulating this pathway could preserve or restore theβcell population in diabetes while cautioning about potential off-target effects(e.g.inαcells).Targeting Rheb1 signaling represents a promising new frontier in diabetes therapy to enhanceβcell resilience;however,a balanced approach addressing both its benefits and risks is essential.This letter discussed the scientific implications and future research directions needed to translate Rheb1 modulation into clinical application for diabetes.
文摘【目的】研究旨在对努比亚山羊大脑富集Ras同源物(Ras homolog enriched in brain,Rheb)基因进行克隆和分析,并构建其真核表达载体,为进一步揭示Rheb对努比亚山羊骨骼肌调控的分子机理奠定基础。【方法】试验采用RT-PCR方法从努比亚山羊背最长肌组织中扩增Rheb基因,经琼脂糖凝胶电泳及测序验证正确后进行生物信息学分析,同时构建该基因真核表达载体,转染细胞后进行实时荧光定量PCR检测来验证所构建载体的正确性。【结果】努比亚山羊Rheb基因编码区序列长度为555 bp,编码184个氨基酸,Rheb蛋白分子式为C 910 H 1456 N 236 O 279 S 6,分子质量为20359.34 u,原子总数为2887,理论等电点为5.93。Rheb蛋白属于稳定的亲水性蛋白,不包含跨膜结构域。蛋白二级结构预测结果显示,努比亚山羊Rheb蛋白中α-螺旋、β-转角、延伸链和无规则卷曲分别占40.76%、7.07%、22.83%和29.35%。构建的真核表达载体pcDNA3.1-Rheb转染山羊骨骼肌细胞后,与空载体组相比,Rheb基因表达量极显著升高(P<0.01)。【结论】本试验成功克隆努比亚山羊Rheb基因编码区序列,并构建pcDNA3.1-Rheb真核表达载体,这为深入理解Rheb基因在努比亚山羊肌肉中的作用提供了理论支持。
基金supported by grants 82170886 and 81800758 from the National Nature Science Foundation of Chinapartially supported by the Fundamental Research Funds for the Central Universities of Central South University(2021zzts1048)。
文摘Dysregulation of microRNAs(miRNAs)in adipocytes plays a critical role in the pathogenesis of obesity.However,the signaling mechanisms regulating miRNAs production in adipose tissue remain largely unclear.Here,we show that adipose tissue-specific knockout of Ras homolog enriched in brain(Rheb),a direct upstream activator of mTOR,increases miR-182-5p level in mouse subcutaneous white adipose tissues.Interestingly,the inhibition of mTOR signaling by rapamycin has no effect on miR-182-5p level in primary subcutaneous white adipocytes,suggesting the presence of a mTOR-independent mechanism regulating Rheb-mediated miR-182-5p expression.Consistent with this view,Rheb-ablation activates the cAMP/PPARγsignaling pathway.In addition,treatment of white adipocytes with pioglitazone,a PPARγagonist,dramatically upregulates miR-182-5p levels.Our study reveals a unique mechanism by which Rheb regulates miR-182-5p in adipocytes.Given that increasing miR-182-5p in adipose tissue promotes beige fat development,our study also suggests a unique mechanism by which Rheb promotes thermogenesis and energy expenditure.
