BaP/BPDE suppressed endothelial cell angiogenesis to induce miscarriage by promoting MARCHF1/GPX4-mediated ferroptosis.

Environmental benzo(a)pyrene (BaP) and its ultimate metabolite BPDE (benzo(a)pyrene-7,8-dihydrodiol-9,10-epoxide) are universal and inevitable persistent organic pollutants and endocrine disrupting chemicals. Angiogenesis in placental decidua plays a pivotal role in healthy pregnancy. Ferroptosis is a newly identified and iron-dependent cell death mode. However, till now, BaP/BPDE exposure, ferroptosis, defective angiogenesis, and miscarriage have never been correlated; and their regulatory mechanisms have been rarely explored. In this study, we used assays with BPDE-exposed HUVECs (human umbilical vein endothelial cells), decidual tissues and serum samples collected from unexplained recurrent miscarriage and their matched healthy control groups, and placental tissues of BaP-exposed mouse miscarriage model. We found that BaP/BPDE exposure caused ferroptosis and then directly suppressed angiogenesis and eventually induced miscarriage. In mechanism, BaP/BPDE exposure up-regulated free Fe2+ level and promoted lipid peroxidation and also up-regulated MARCHF1 (a novel E3 ligase of GPX4) level to promote the ubiquitination degradation of GPX4, both of which resulted in HUVEC ferroptosis. Furthermore, we also found that GPX4 protein down-regulated the protein levels of VEGFA and ANG-1, two key proteins function for angiogenesis, and thus suppressed HUVEC angiogenesis. In turn, supplement with GPX4 could suppress ferroptosis, recover angiogenesis, and alleviate miscarriage. Moreover, the levels of free Fe2+ and VEGFA in serum might predict the risk of miscarriage. Overall, this study uncovered the crosstalk among BaP/BPDE exposure, ferroptosis, angiogenesis, and miscarriage, discovering novel toxicological effects of BaP/BPDE on human reproductive health. This study also warned the public to avoid exposure to polycyclic aromatic hydrocarbons during pregnancy to effectively prevent adverse pregnancy outcomes.

Comparative analysis of flavonoids in white and red table grape cultivars during ripening by widely targeted metabolome and transcript levels.

The flavonoid metabolites were compared between red 'Summer Black' (SB) and white 'Shine Muscat' (SM) table grapes during fruit development based on widely targeted metabolome. A total of 134 flavonoids were identified in two cultivars, including 37 flavones, 33 flavonols, and 11 anthocyanidins, and so on. From young to veraison, the composition and the content of most flavonoids were decreasing in both cultivars but increased at maturation in SB. In general, SB has higher flavonoid compositions and content than SM during the whole fruit development, especially the content of anthocyanin after veraison. While the SM had higher content of flavonols such as quercetin, kaempferol and their derivatives. The expression of anthocyanin-related genes such as UFGT, OMT, GST, MATE, MYBA1, and MYBA2 was remarkably higher in SB than those in SM, which may attribute to higher anthocyanin content, while the higher expression of F3H and FLS resulted higher level of flavonols in SM. These results improve our understanding of flavonoid profiles and molecular mechanism in table grape cultivars.

Methylation of MYBA1 is Associated with the Coloration in "Manicure Finger" Grape Skin.

The "Manicure Finger" grape is notable for its fingerlike berries with a bright red top and yellow base; however, the mechanism underlying this color difference remains unknown. This study showed that the anthocyanin concentration and the expression levels of anthocyanin-related genes in the top skin were notably higher than those in the basal skin. The expression levels of DFR, UFGT, and GST were significantly correlated with the anthocyanin content. The promoters of the two VvUFGT alleles can be activated by VvMYBA1, which was verified by the yeast one-hybrid assay, the dual-luciferase reporter gene assay, and the electrophoretic mobility shift assay. Moreover, the methylation level of the VvMYBA1 promoter (-1488 to -1083 bp) in the top skin was significantly lower than that in the basal skin and was positively correlated with the anthocyanin content. Our data suggest that methylation levels of the VvMYBA1 promoter play a crucial role in regulating grape skin coloration.

Melatonin application improves berry coloration, sucrose synthesis, and nutrient absorption in 'Summer Black' grape.

In this study, we investigated the effects of melatonin application on berry coloration, sugar accumulation, and nutrient absorption in 'Summer Black' grapes. Melatonin spraying at 100 µmol L-1 on grapes during veraison induced skin coloration earlier than that in controls, as well as higher transcript abundance of anthocyanin biosynthesis-related genes and transcription factors MYBA1 and MYBA2. Melatonin treatment increased the soluble sugar content, especially of sucrose, by promoting the activity of sucrose phosphate synthase, and also increased endogenous melatonin content and the concentrations of mineral nutrients N, K, Cu, Fe, and Zn in grape berries. Correlation analysis suggested that high sugar content promoted anthocyanin synthesis. These findings provide a sound theoretical basis for the development of techniques aimed to achieve optimum coloration of grapes in hot and rainy regions.

Sp1-mediated upregulation of Prdx6 expression prevents podocyte injury in diabetic nephropathy via mitigation of oxidative stress and ferroptosis.

Glomerular podocyte damage is considered to be one of the main mechanisms leading to Diabetic nephropathy (DN). However, the relevant mechanism of podocyte injury is not yet clear. This study aimed to investigate the effect of peroxiredoxin 6 (Prdx6) on the pathogenesis of podocyte injury induced by high glucose (HG). The mouse glomerular podocyte MPC5 was stimulated with 30 nM glucose, and the Prdx6 overexpression vector or specificity protein 1 (Sp1) overexpression vector was transfected into MPC5 cells before the high glucose stimulation. As results, HG treatment significantly reduced the expression of Prdx6 and Sp1 in MPC5 cells. Prdx6 overexpression increased cell viability, while inhibited podocyte death, inflammation and podocyte destruction in HG-induced MPC5 cells. Prdx6 overexpression inhibited HG-induced ROS and MDA production, while restored SOD and GSH activity in MPC5 cells. Prdx6 overexpression also eliminated ferroptosis caused by HG, which was reflected in the suppression of iron accumulation and the increase in SLC7A11 and GPX4 expression. The improvement effect of Prdx6 on HG-induced podocyte damage could be eliminated by erastin. Moreover, Sp1 could bind to the three Sp1 response elements in the Prdx6 promoter, thereby directly regulating the transcriptional activation of Prdx6 in podocytes. Silencing Sp1 could eliminate the effect of Prdx6 on HG-induced podocyte damage. Further, Prdx6 overexpression attenuated renal injuries in streptozotocin-induced DN mice. Sp1-mediated upregulation of Prdx6 expression prevents podocyte injury in diabetic nephropathy via mitigation of oxidative stress and ferroptosis, which may provide new insights for the study of the mechanism of DN.

Melatonin Accumulation in Sweet Cherry and Its Influence on Fruit Quality and Antioxidant Properties.

Although the effects of melatonin on plant abiotic and biotic stress resistance have been explored in recent decades, the accumulation of endogenous melatonin in plants and its influence on fruit quality remains unclear. In the present study, melatonin accumulation levels and the expression profiles of five synthesis genes were investigated during fruit and leaf development in sweet cherry (Prunus avium L.). Melatonin was strongly accumulated in young fruits and leaves, then decreased steadily with maturation. Transcript levels of PacTDC and PacSNAT were highly correlated with melatonin content in both fruit and leaves, indicating their importance in melatonin accumulation. Furthermore, application of 50 and 100 µmol·L-1 of melatonin to leaves had a greater influence on fruit quality than treatments applied to fruits, by significantly improving fruit weight, soluble solids content, and phenolic content including total phenols, flavanols, total anthocyanins, and ascorbic acid. Meanwhile, melatonin application promoted the antioxidant capacity of fruit assayed by 2,2-diphenyl-1-picrylhydrazyl (DPPH), 2,2'-azinobis (3-ethylben zothiazoline-6-sulfonic acid) (ABTS), and ferric reducing antioxidant power (FRAP). These results provide insights into the physiological and molecular mechanisms underlying melatonin metabolism of sweet cherry.

Hydrogen cyanamide induces grape bud endodormancy release through carbohydrate metabolism and plant hormone signaling.

BACKGROUND: Grape buds exhibit non-uniform, or delayed, break in early spring in subtropical regions because the accumulation of chilling is insufficient. Hydrogen cyanamide (H2CN2, HC) can partially replace chilling to effectively promote bud sprouting and is used widely in warm winter areas. However, the exact underlying mechanism of grape bud release from endodormancy induced by HC remains elusive. RESULTS: In this study, the transcriptome of grape winter buds under in vitro conditions following HC and water treatment (control) was analyzed using RNA-seq technology. A total of 6772 differentially expressed genes (DEGs) were identified. Furthermore, the gene ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) analysis revealed that starch and sucrose metabolism and plant hormone signaling transduction were most enriched out of the 50 total pathways. HC treatment induced the upregulated expression of sucrose synthase (SUS), sucrose phosphate synthase (SPS), α-amylase (AM), and ß-amylase (BM) and downregulated expression of sucrose invertase (INV), hexokinase (HK), fructokinase (FK), soluble starch synthase (SS), and granule-bound starch synthase (GBSS). Hence, the starch concentration in the HC-treated group was significantly lower than that in control, whereas soluble sugar content in the HC-treated group increased quickly and was higher than that in control between 0 and 8 d. The concentration of indoleacetic acid (IAA) and zeatin (ZT) increased, whereas that of abscisic acid (ABA) and gibberellin (GA) decreased in HC treated group, which coincided with the expression level of genes involved in above hormone signals. The content of hydrogen peroxide (H2O2) and enzyme activity of superoxide dismutase (SOD) and peroxidase (POD) were increased in grape buds with HC treatment, whereas catalase (CAT) activity was decreased. HC treatment increased the expression of POD, SOD, primary amine oxidase (PAO), polyamine oxidase (PAOX), and glutathione peroxidase (GSH-Px). CONCLUSION: Based on these results, it is possible to propose a mechanistic model that underlies the regulation of endodormancy release in grapevine buds by exogenous HC application.

Exogenous Melatonin Application Delays Senescence of Kiwifruit Leaves by Regulating the Antioxidant Capacity and Biosynthesis of Flavonoids.

Melatonin, a multiple signal molecule, plays important roles in delaying senescence during the development of plants. Because few species have been studied for the effect of exogenous melatonin on anti-aging, the plausible mechanism of melatonin of anti-aging effects on other plant species has remained largely unknown. In the present study, the effects of exogenous melatonin on leaf senescence in kiwifruit were examined during natural aging after melatonin (200 µM) or water (Control) pretreatment. The decreased membrane damage and lower hydrogen peroxide (H2O2) content due to the enhanced scavenging activity of antioxidant enzymes peroxidase (POD), superoxide dismutase (SOD), and catalase (CAT) demonstrated that melatonin effectively delayed the aging of kiwifruit leaves. Likewise, owing to up-regulated expression of chlorophyll a/b-binding protein (CAB) gene in the sampled leaves pretreated with melatonin, chlorophyll degradation decreased. Therefore, osmoregulatory substances in sampled leaves accumulated (e.g., soluble sugar and soluble protein) and seedling cell environment stability was maintained. Simultaneously, melatonin decreased H2O2 concentration owing to increased glutathione (GSH) and ascorbate (AsA) content, and the expression levels of glutathione reductase (GR), ascorbate peroxidase (APX), monodehydroascorbate reductase (MDAR), dehydroascorbate reductase (DHAR) were up-regulated by melatonin application, indicating that the increase of GSH and AsA was attributed to the expression of these genes. In addition, a large amount of flavonoids accumulated in seedlings pretreated with melatonin, and transcript levels of eight genes involved in flavonoid synthesis, including phenylalanine ammonia-lyase (PAL), cinnamate-4-hydroxymate (C4H), chalcone synthase (CHS), flavanone 3-hydroxylase (F3H), flavonol synthase (FNS), leucoanthocyanin reductase (LAR), anthocyanin reductase (ANR), flavonoid 3-O-glucosyltransferase (UFGT) were enhanced in response to melatonin application. These results indicated that melatonin delayed aging of kiwifruit leaves by activating the antioxidant capacity and enhancing flavonoid biosynthesis. All of these results can provide clear proof that melatonin plays a key roles in delaying leaf senescence.