Search

• A fluorescent probe, leonurine-P, was facilely prepared from coupling FITC and the small-molecule phytodrug leonurine. • Leonurine-P can be used to visualize spatiotemporal dynamics processes such as drug uptake and distribution in C. elegans and living cells. • Leonurine-P can be further explored in standard fluorescence imaging or flow cytometric platforms to allow quantitative analyses. Caenorhabditis elegans (C. elegans) has been recognized for being a useful model organism in small-molecule drug screens and drug efficacy investigation. However, there remain bottlenecks in evaluating such processes as drug uptake and distribution due to a lack of appropriate chemical tools. This study aims to prepare fluorescence-labeled leonurine as an example to monitor drug uptake and distribution of small molecule in C. elegans and living cells. FITC-conjugated leonurine (leonurine-P) was synthesized and characterized by LC/MS, NMR, UV absorption and fluorescence intensity. Leonurine-P was used to stain C. elegans and various mammalian cell lines. Different concentrations of leonurine were tested in conjunction with a competing parent molecule to determine whether leonurine-P and leonurine shared the same biological targets. Drug distribution was analyzed by imaging. Fluorometry in microplates and flow cytometry were performed for quantitative measurements of drug uptake. The UV absorption peak of leonurine-P was 490∼495 nm and emission peak was 520 nm. Leonurine-P specifically bound to endogenous protein targets in C. elegans and mammalian cells, which was competitively blocked by leonurine. The highest enrichment levels of leonurine-P were observed around 72 h following exposure in C. elegans. Leonurine-P can be used in a variety of cells to observe drug distribution dynamics. Flow cytometry of stained cells can be facilely carried out to quantitatively detect probe signals. The strategy of fluorescein-labeled drugs reported herein allows quantification of drug enrichment and visualization of drug distribution, thus illustrates a convenient approach to study phytodrugs in pharmacological contexts. [Display omitted] [ABSTRACT FROM AUTHOR]

• ZXT extend lifespan and stress resistance in C. elegans. • ZXT alleviates fat accumulation induced by high sugar by regulating fat synthesis and catabolism. • ZXT attenuated oxidative stress by preventing mitochondrial dysfunction via skn-1 and increasing the expression of sod-3 and daf-16. Obesity has become a global public health problem. Zexie decoction (ZXT) is a classic formula from Synopsis of the Golden Chamber. However, the long-term effect of ZXT in lipid accumulation remain unclear. This study aims to investigate the effect of ZXT on aging, lipid metabolism and oxidative stress. Different concentration of ZXT was administered to Caenorhabditis elegans (C. elegans) cultured in NGM or the high glucose nematode growth media (GNGM). The lifespan, heat stress resistance, lipid accumulation and related mRNA expression of the worms were examined. Oil Red staining and triglyceride were used to evaluated the lipid accumulation. Nhr-49, fat-5/fat-7, fat-5/fat-6 or skn-1 knockout mutants were used to clarify the effect on lipid metabolism of ZXT. GFP-binding mutants were used to observe the changes in protein expression. ZXT improved the survival rate of C. elegans in lifespan test and heat stress test. ZXT also reduced lipid accumulation in C. elegans and significantly changed the expression of fatty acid synthesis related genes and lipid metabolism related genes. In addition, ZXT-treated C. elegans showed a higher expression of anti-oxidative protein, and reduced the expression of oxidative stress and mitochondrial dysfunction marker. However, when skn-1 was knockdown, ZXT no longer had the effect of maintaining the mitochondria membrane potential and lipid lowering but still effectively decreased the O 2 ·− induced by high glucose. ZXT reduced fat accumulation by regulating lipid metabolism via multiple targets and enhanced stress resistance by its antioxidant effect in C. elegans. [Display omitted] [ABSTRACT FROM AUTHOR]

Cadmium (Cd) migration in the rhizosphere soil is easily affected by plants and microorganisms. Global warming significantly affects plant growth, and arbuscular mycorrhizal fungi (AMF) can chelate heavy metals by mycelium, cell wall components, and mycelial secretion. Here, we investigated the regulation of Glomus mosseae on Cd migration in the rhizosphere soil of alfalfa under elevated temperature (ET, + 3 °C). Elevated temperature significantly decreased G. mosseae colonization rate in the roots by 49.5% under Cd exposure. Under ET + G. mosseae + Cd relative to ET + Cd, the contents of free amino acids, total and easily extractable glomalin-related soil protein (GRSP), and root Cd increased significantly; however, the changes in DTPA-Cd in the rhizosphere soil and Cd in the shoots were insignificant. In addition, G. mosseae colonization enhanced the bioconcentration factor of Cd in the roots and the total removal rate of Cd in the rhizosphere soil by 63.4% and 16.3%, respectively, under ET + Cd. However, the changes in the expression of iron-regulated transport 1 (IRT1) and natural resistance-associated macrophage protein 1 genes were insignificant under ET + G. mosseae + Cd relative to ET + Cd. In summary, temperature and G. mosseae significantly affected Cd fate in the rhizosphere soil, and IRT1 gene and rhizosphere soil pH, N, and C/N ratio were significant factors influencing Cd migration. Additionally, G. mosseae improved the remediation efficiency of Cd-contaminated soils by alfalfa under ET. The results will help us understand the regulation of AMF on the phytoremediation of heavy metal-contaminated soils under global warming scenarios. • Elevated temperature (ET, + 3 °C) reduced Glomus mosseae colonization under Cd. • G. mosseae significantly improved GRSP content in the rhizosphere soil under ET + Cd. • G. mosseae had no significant effect on IRT1 and NRAMP 1 gene expression. • G. mosseae significantly enhanced Cd uptake by the roots of alfalfa under ET. • Rhizosphere soil pH, C, C/N ratio and IRT1 gene significantly affected Cd fate. [ABSTRACT FROM AUTHOR]

Arbuscular mycorrhizal fungi (AMF) are easily influenced by increasing atmospheric CO 2 concentration and heavy metals including cadmium (Cd), which can regulate antioxidant enzyme in host plants. Although the effect of AMF under individual conditions such as elevated CO 2 (ECO 2) and Cd on antioxidant enzyme in host plants has been reported widely, the effect of AMF under ECO 2 + Cd receives little attention. In this study, a pot experiment was conducted to study the effect of AMF community in roots on superoxide dismutase (SOD), catalase (CAT), and peroxidase (POD) activities in leaves of 135-d Robinia pseudoacacia L. seedlings under ECO 2 + Cd. The activities of SOD and CAT increased and POD activity and the richness and diversity of AMF community decreased under ECO 2 + Cd relative to Cd alone. The richness and diversity of AMF were negatively related to Cd content in roots and leaves. The richness and OTUs of AMF community positively and AMF gene abundance negatively affected POD activity under the combined treatments. Superoxide dismutase and POD activities were negatively and positively related to Archaeospora and Scutellospora , respectively, under ECO 2 + Cd. Cadmium in roots and leaves was negatively and significantly related to Glomus , Scutellospora , and Claroideoglomus abundance under ECO 2 + Cd. Overall, AMF diversity and Archaeospora and Scutellospora in roots significantly influenced SOD, POD, and CAT activities. The response of AM symbiosis to ECO 2 might regulate antioxidant capacity in host plants upon Cd exposure. Glomus , Scutellospora , and Claroideoglomus might be applied to phytoremediation of Cd-contaminated soils. [Display omitted] • Decrease in AMF richness stimulated POD activity under elevated CO 2 + Cd. • AMF community in roots significantly affected SOD, CAT, and POD activities. • POD activity was positively related (p < 0.05) to Archaeospora and Scutellospora. • SOD activity was negatively related (p < 0.05) to Archaeospora and Scutellospora. • Glomus , Scutellospora , and Claroideoglomus might be applied to phytoremediation. [ABSTRACT FROM AUTHOR]