AMPK activation protects cells from oxidative stress‐induced senescence via autophagic flux restoration and intracellular NAD+ elevation

AMPK activation is beneficial for cellular homeostasis and senescence prevention. However, the molecular events involved in AMPK activation are not well defined. In this study, we addressed the mechanism underlying the protective effect of AMPK on oxidative stress‐induced senescence. The results showed that AMPK was inactivated in senescent cells. However, pharmacological activation of AMPK by metformin and berberine significantly prevented the development of senescence and, accordingly, inhibition of AMPK by Compound C was accelerated. Importantly, AMPK activation prevented hydrogen peroxide‐induced impairment of the autophagic flux in senescent cells, evidenced by the decreased p62 degradation, GFP‐RFP‐LC3 cancellation, and activity of lysosomal hydrolases. We also found that AMPK activation restored the NAD⁺ levels in the senescent cells via a mechanism involving mostly the salvage pathway for NAD⁺ synthesis. In addition, the mechanistic relationship of autophagic flux and NAD⁺ synthesis and the involvement of mTOR and Sirt1 activities were assessed. In summary, our results suggest that AMPK prevents oxidative stress‐induced senescence by improving autophagic flux and NAD⁺ homeostasis. This study provides a new insight for exploring the mechanisms of aging, autophagy and NAD⁺ homeostasis, and it is also valuable in the development of innovative strategies to combat aging.     

Functional TRPV2 and TRPV4 channels in human cardiac c‐kit+ progenitor cells

The cellular physiology and biology of human cardiac c‐kit⁺ progenitor cells has not been extensively characterized and remains an area of active research. This study investigates the functional expression of transient receptor potential vanilloid (TRPV) and possible roles for this ion channel in regulating proliferation and migration of human cardiac c‐kit⁺ progenitor cells. We found that genes coding for TRPV2 and TRPV4 channels and their proteins are significantly expressed in human c‐kit⁺ cardiac stem cells. Probenecid, an activator of TRPV2, induced an increase in intracellular Ca²⁺ (Ca²⁺_i), an effect that may be attenuated or abolished by the TRPV2 blocker ruthenium red. The TRPV4 channel activator 4α‐phorbol 12‐13‐dicaprinate induced Ca²⁺_i oscillations, which can be inhibited by the TRPV4 blocker RN‐1734. The alteration of Ca²⁺_i by probenecid or 4α‐phorbol 12‐13‐dicprinate was dramatically inhibited in cells infected with TRPV2 short hairpin RNA (shRNA) or TRPV4 shRNA. Silencing TRPV2, but not TRPV4, significantly reduced cell proliferation by arresting cells at the G0/G1 boundary of the cell cycle. Cell migration was reduced by silencing TRPV2 or TRPV4. Western blot revealed that silencing TRPV2 decreased expression of cyclin D1, cyclin E, pERK1/2 and pAkt, whereas silencing TRPV4 only reduced pAkt expression. Our results demonstrate for the first time that functional TRPV2 and TRPV4 channels are abundantly expressed in human cardiac c‐kit⁺ progenitor cells. TRPV2 channels, but not TRPV4 channels, participate in regulating cell cycle progression; moreover, both TRPV2 and TRPV4 are involved in migration of human cardiac c‐kit⁺ progenitor cells.     

A large‐scale phylogeny of the lycophyte genus Selaginella (Selaginellaceae: Lycopodiopsida) based on plastid and nuclear loci

The lycophyte genus Selaginella alone constitutes the family Selaginellaceae, the largest of the lycophyte families. The genus is estimated to contain 700–800 species distributed on all continents except Antarctica, with highest species diversity in tropical and subtropical regions. The monophyly of Selaginella in this broad sense has rarely been doubted, whereas its intrageneric classification has been notoriously contentious. Previous molecular studies were based on very sparse sampling of Selaginella (up to 62 species) and often used DNA sequence data from one genome. In the present study, DNA sequences of one plastid (rbcL) and one nuclear (ITS) locus from 394 accessions representing approximately 200 species of Selaginella worldwide were used to infer a phylogeny using maximum likelihood, Bayesian inference and maximum parsimony methods. The study identifies strongly supported major clades and well resolves relationships among them. Major results include: (i) six deep‐level clades are discovered representing the deep splits of Selaginella; and (ii) 20 major clades representing 20 major evolutionary lineages are identified, which differ from one another in molecular, macro‐morphological, ecological and spore features, and/or geographical distribution.     

红外光谱结合多元统计方法的不同产地红根草红外指纹图谱比较研究

红根草为唇形科鼠尾带根全草植物,是著名的广西道地药材和常用中药,对白血病细胞有很强的抑制作用,同时具有较强的抗菌活性和抗癌作用,主治菌莉、腹泻、肠炎、肺炎、急性咽喉炎、扁桃体炎、感冒等症。为快速鉴别和评价不同产地中药红根草主要化学成分的差异,该研究利用红外光谱对不同产地红根草进行检测,并结合主成分分析和聚类分析及载荷因子等方法对不同产地样本进行鉴别。结果表明：(1)在1800~600 cm-1范围内,不同产地红根草根系在1727、1635、1551、1513、1442、1373、1255、1154、1036、795、776、690 cm-1等处均有较强的振动吸收,表明不同产地红根草主要化学组分构成比较相似。(2)红外指纹图谱结合主成分和聚类分析结果表明,不同产地红根草化学成分的差异与地理位置有明显对应性,产地相近的地区红根草化学成分的较似,产地较远的区域红根草化学成分差异较大,但两种方法检测结果均有自己的特征。(3)通过PCA载荷因子分析,可以得出比原始图谱更多的化学成分信息,对主成分聚类贡献较大的吸收峰主要表现在1670、1630、1616、1579、1473、1411、1159、1129、1082、1042、1000、972、946、913、891、806 cm-1附近,进一步揭示出不同产地红根草化学成分差异主要是红根草内酯和甾醇类成分,以及主要有效成分红根草邻醌和丹参酮类成分的差异。该研究结果为红根草的引种栽培及良种选育研究提供了参考。     

非ST段抬高型急性冠脉综合征患者血浆S100A1水平与GRACE评分及短期预后相关性的研究

目的:探讨非ST段抬高型急性冠脉综合征(NST-ACS)患者血浆S100A1水平与全球急性冠状动脉事件注册(GRACE)评分之间的关系,以及S100A1水平对NST-ACS患者30天预后的判定价值。方法:共有162例NST-ACS患者符合入选标准,收集基本临床资料,进行GRACE评分,同时收集次日清晨空腹采集肘静脉血,检测血浆S100A1浓度,与患者的GRACE评分进行比较。根据S100A1的水平进行分组随访,KM生存分析不同组患者30天预后进行评价。结果:不同GRACE分组患者间S100A1水平具有显著性差异(P0.05);相关性分析显示,NST-ACS患者S100A1与GRACE评分呈显著正相关(r=0.49,P0.01);KM生存分析显示,S100A1水平3.41 ng/mL的患者30天内心血管事件发生率显著升高(P0.05)。结论:S100A1可作为预测NST-ACE患者病情的发生发展的生化指标;在NST-ACS患者中运用S100A1有助于对患者早期危险分层及评估预后有一定的临床价值。