Caloric restriction confers persistent anti-oxidative, pro-angiogenic, and anti-inflammatory effects and promotes anti-aging miRNA expression profile in cerebromicrovascular endothelial cells of aged rats
Caloric restriction confers persistent anti-oxidative, pro-angiogenic, and anti-inflammatory effects and promotes anti-aging miRNA expression profile in cerebromicrovascular endothelial cells of aged rats
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DOI：
10.1152/ajpheart.00307.2014
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发表时间：
2014-08-01
影响因子：
4.8
通讯作者：
Ungvari, Zoltan
中科院分区：
文献类型：
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作者：
Csiszar, Anna;Gautam, Tripti;Ungvari, Zoltan
在啮齿动物中，无营养不良的适度热量限制(CR)具有显著的脑血管保护作用，改善了皮质微血管密度和内皮依赖性血管扩张，但其潜在的细胞机制仍不清楚。为探讨CR F344xBN对脑微血管内皮细胞(CMVECs)的维持作用，从自由喂养的青年(3月龄)和老年(24月龄)大鼠和老年CRF344xBN大鼠分离出原代CMVECs。我们发现，与年龄相关的细胞和线粒体氧化应激增加，这是由CR防止的。在老年的CMVECs中，Nrf2的表达和转录活性均显著降低，而CR可防止与年龄相关的Nrf2功能障碍。在老年CMVECs中miR-144的表达上调，而在幼年动物和老年CR大鼠来源的细胞中miR-144的过度表达显著降低Nrf2的表达。在衰老的CMVEC中过表达miR-144抑制因子显著降低miR-144的表达并上调Nrf2的表达。我们发现，CR可防止与年龄相关的血管生成过程的损伤，包括细胞增殖、与胶原的黏附和毛细血管样结构的形成，并抑制CMVECs的凋亡。CR还具有显著的抗炎作用，防止与年龄相关的核因子-kappaB转录活性的增加和与年龄相关的内皮分泌组的促炎转移。CR诱导的miRNA表达变化的特征表明，它们可能影响内皮细胞动态平衡的几个关键功能。预测的CR相关差异表达的miRNAs对老年CMVECs的调节作用与体内观察到的CR的抗衰老内皮效应是一致的。总而言之，我们发现CR具有持久的抗氧化、促血管生成和抗炎细胞作用，保留了大鼠脑微血管内皮细胞的年轻表型，提示CR可能通过这些作用改善脑血管功能和预防血管认知障碍。
In rodents, moderate caloric restriction (CR) without malnutrition exerts significant cerebrovascular protective effects, improving cortical microvascular density and endothelium-dependent vasodilation, but the underlying cellular mechanisms remain elusive. To elucidate the persisting effects of CR on cerebromicrovascular endothelial cells (CMVECs), primary CMVECs were isolated from young (3 mo old) and aged (24 mo old) ad libitum-fed and aged CR F344xBN rats. We found an age-related increase in cellular and mitochondrial oxidative stress, which is prevented by CR. Expression and transcriptional activity of Nrf2 are both significantly reduced in aged CMVECs, whereas CR prevents age-related Nrf2 dysfunction. Expression of miR-144 was upregulated in aged CMVECs, and overexpression of miR-144 significantly decreased expression of Nrf2 in cells derived from both young animals and aged CR rats. Overexpression of a miR-144 antagomir in aged CMVECs significantly decreases expression of miR-144 and upregulates Nrf2. We found that CR prevents age-related impairment of angiogenic processes, including cell proliferation, adhesion to collagen, and formation of capillary-like structures and inhibits apoptosis in CMVECs. CR also exerts significant anti-inflammatory effects, preventing age-related increases in the transcriptional activity of NF-kappa B and age-associated pro-inflammatory shift in the endothelial secretome. Characterization of CR-induced changes in miRNA expression suggests that they likely affect several critical functions in endothelial cell homeostasis. The predicted regulatory effects of CR-related differentially expressed miRNAs in aged CMVECs are consistent with the anti-aging endothelial effects of CR observed in vivo. Collectively, we find that CR confers persisting anti-oxidative, pro-angiogenic, and anti-inflammatory cellular effects, preserving a youthful phenotype in rat cerebromicrovascular endothelial cells, suggesting that through these effects CR may improve cerebrovascular function and prevent vascular cognitive impairment.