Antioxidant and anti-aging activities of polysaccharides from Cordyceps cicadae.
Cordyceps cicadae is a traditional Chinese medicine with high nutritional value and biological activities. Previously, we reported on the antioxidant activity associated with the polysaccharides from Cordyceps cicadae (CP). To further explore which of the fraction of CP had the greatest potency, in here, the in vitro antioxidant and in vivo anti-aging activities of the fractions CP30-CP80 of CP were evaluated. The in vitro antioxidant activity results revealed that all the fractions (i.e. CP30-CP80) were potent with CP70 as the most potent. Notably, CP70 prolonged the lifespan of Drosophila (P < 0.05), increased the activities of catalase (CAT) and glutathione peroxidase (GSH-Px) (P < 0.01), and inhibited the formation of malondialdehyde (MDA) (P < 0.01). Additionally, CP70 upregulated the expression level of antioxidant-related genes CAT, SOD1 and MTH in Drosophila (P < 0.05). These results indicated that CP70 may prolong the lifespan of Drosophila through the up-regulation of the expression level of antioxidant-related genes CAT, SOD1 and MTH in Drosophila. Thus, polysaccharides from Cordyceps cicadae possess significant antioxidant and anti-aging activities, and could be explored as a new dietary supplement to slow down the aging process.
miR-129-5p: A key factor and therapeutic target in amyotrophic lateral sclerosis.
Amyotrophic lateral sclerosis (ALS) is a relentless and fatal neurological disease characterized by the selective degeneration of motor neurons. No effective therapy is available for this disease. Several lines of evidence indicate that alteration of RNA metabolism, including microRNA (miRNA) processing, is a relevant pathogenetic factor and a possible therapeutic target for ALS. Here, we showed that the abundance of components in the miRNA processing machinery is altered in a SOD1-linked cellular model, suggesting consequent dysregulation of miRNA biogenesis. Indeed, high-throughput sequencing of the small RNA fraction showed that among the altered miRNAs, miR-129-5p was increased in different models of SOD1-linked ALS and in peripheral blood cells of sporadic ALS patients. We demonstrated that miR-129-5p upregulation causes the downregulation of one of its targets: the RNA-binding protein ELAVL4/HuD. ELAVL4/HuD is predominantly expressed in neurons, where it controls several key neuronal mRNAs. Overexpression of pre-miR-129-1 inhibited neurite outgrowth and differentiation via HuD silencing in vitro, while its inhibition with an antagomir rescued the phenotype. Remarkably, we showed that administration of an antisense oligonucleotide (ASO) inhibitor of miR-129-5p to an ALS animal model, SOD1 (G93A) mice, result in a significant increase in survival and improved the neuromuscular phenotype in treated mice. These results identify miR-129-5p as a therapeutic target that is amenable to ASO modulation for the treatment of ALS patients.
PACAP Modulates the Autophagy Process in an In Vitro Model of Amyotrophic Lateral Sclerosis.
Amyotrophic lateral sclerosis (ALS) is a progressive neurodegenerative disease of complex etiology leading to motor neuron degeneration. Many gene alterations cause this pathology, including mutation in Cu, Zn superoxide dismutase (SOD1), which leads to its gain of function. Mutant SOD1 proteins are prone to aberrant misfolding and create aggregates that impair autophagy. The hypoxic stress is strictly linked to the disease progression since it induces uncontrolled autophagy activation and the consequent high rates of cell death. Previously, we showed that pituitary adenylate cyclase-activating polypeptide (PACAP) exerts neurotrophic activity in cultured mSOD1 motor neurons exposed to serum deprivation. To date, no studies have examined whether the protective effect of PACAP on mSOD1 cells exposed to hypoxic insult is mediated through the regulation of the autophagy process. In the present study, we used the neuroblastoma-spinal cord-34 (NSC-34) cell line, stably expressing human wild type or mutant SOD1 G93A, to represent a well characterized in vitro model of a familial form of ALS. These cells were exposed to 100-µM desferrioxamine mesylate salt for 24h, to mimic the hypoxic stress affecting motor neurons during the disease progression. Our results showed that PACAP treatment significantly reduced cell death and hypoxia-induced mSOD1 accumulation by modulating the autophagy process in G93A motor neurons, as revealed by the decreased LC3II and the increased p62 levels, two autophagy indicators. These results were also confirmed by evaluating the vacuole formation detected through light chain 3 (LC3) immunofluorescence. Furthermore, the PACAP effects on autophagy seem to be mediated through the activation of the MAPK/ERK signaling pathway. Overall, our data demonstrated that PACAP exerts an ameliorative effect on the mSOD1 motor neuron viability by modulating a hypoxia-induced autophagy process through activation of MAPK/ERK signaling cascade.
Reactive oxygen species triggers unconventional secretion of antioxidants and Acb1.
Nutrient deprivation triggers the release of signal-sequence-lacking Acb1 and the antioxidant superoxide dismutase 1 (SOD1). We now report that secreted SOD1 is functionally active and accompanied by export of other antioxidant enzymes such as thioredoxins (Trx1 and Trx2) and peroxiredoxin Ahp1 in a Grh1-dependent manner. Our data reveal that starvation leads to production of nontoxic levels of reactive oxygen species (ROS). Treatment of cells with N-acetylcysteine (NAC), which sequesters ROS, prevents antioxidants and Acb1 secretion. Starved cells lacking Grh1 are metabolically active, but defective in their ability to regrow upon return to growth conditions. Treatment with NAC restored the Grh1-dependent effect of starvation on cell growth. In sum, starvation triggers ROS production and cells respond by secreting antioxidants and the lipogenic signaling protein Acb1. We suggest that starvation-specific unconventional secretion of antioxidants and Acb1-like activities maintain cells in a form necessary for growth upon their eventual return to normal conditions.
Prx II reduces oxidative stress and cell senescence in chondrocytes by activating the p16-CDK4/6-pRb-E2F signaling pathway.
Osteoarthritis (OA) is a common clinical degenerative disease and has a high incidence in the elderly. The purpose of this study was to explore the anti-oxidative stress and anti-aging effects of Peroxiredoxin II (Prx II) on articular chondrocytes, as well as its molecular mechanism.Articular cartilage tissues and culture human articular chondrocytes were selected. By constructing Prx II overexpressing lentivirus, the effects of Prx II on oxidative stress and cell senescence in chondrocytes were studied. Besides, the p16 overexpression lentivirus was constructed to investigate the effect of Prx II on the p16-CDK4/6-pRb-E2F signaling pathway (p16 signaling pathway).Articular cartilage tissues in patients with OA and IL-1β-induced chondrocytes expressed lower Prx II and had higher p16 signaling pathway activity. The overexpression of Prx II significantly increased the expression of SOD1 and SOD2 and decreased the expression of β-gal and P53/P21, indicating that Prx II can reduce the oxidative stress and senescence level of chondrocytes. Moreover, the overexpression of Prx II increased the expression of p16 signaling pathway-related molecules and the activation of the p16 signaling pathway attenuated the anti-oxidative stress and anti-aging effects of Prx II.Prx II can inhibit the p16 signaling pathway in chondrocytes to reduce the level of aging in chondrocytes, thereby reducing the level of oxidative stress in chondrocytes, and ultimately inhibiting the progression of OA.
Quercetin supports bovine preimplantation embryo development under oxidative stress condition via activation of the Nrf2 signaling pathway.
Nrf2 is a master regulator for antioxidant machinery against oxidative stress in bovine preimplantation embryos. The endogenous or exogenous modulation of Nrf2-KEAP1 system in bovine embryos may contribute to the understanding of the mechanisms behind the response of embryos to stress conditions. Therefore, here we aimed to investigate the protective effect of quercetin on bovine preimplantation embryos exposed to higher atmospheric oxygen concentration. For that, blastocysts, which were developed from zygotes cultured in media supplemented with or without quercetin under high oxygen level (20%), were subjected intracellular ROS level and mitochondrial analysis, and determining blastocyst formation rate and total cell number. Moreover, mRNA and protein expression level of Nrf2 and selected downstream antioxidant genes were investigated in the resulting blastocysts. Quercetin supplementation in vitro culture did not affect cleavage and blastocyst rate until day 7. However, quercetin supplementation resulted in higher blastocyst total cell number and reduction of intracellular ROS level accompanied by increasing mitochondrial activity compared to control group in both day 7 and day 8 blastocysts. Moreover, quercetin supplementation induced mRNA and protein of Nrf2 with subsequent increase in the expression of downstream antioxidants namely: NQO1, PRDX1, CAT, and SOD1 antioxidants. In conclusion, quercetin protects preimplantation embryos against oxidative stress and improves embryo viability through modulation of the Nrf2 signaling pathway.
Receptor-mediated endocytosis 8 (RME-8)/DNAJC13 is a novel positive modulator of autophagy and stabilizes cellular protein homeostasis.
The cellular protein homeostasis (proteostasis) network responds effectively to insults. In a functional screen in C. elegans, we recently identified the gene receptor-mediated endocytosis 8 (rme-8; human ortholog: DNAJC13) as a component of the proteostasis network. Accumulation of aggregation-prone proteins, such as amyloid-β 42 (Aβ), α-synuclein, or mutant Cu/Zn-superoxide dismutase (SOD1), were aggravated upon the knockdown of rme-8/DNAJC13 in C. elegans.
