Astaxanthin protected against the adverse effects induced by diesel exhaust particulate matter via improving membrane stability and anti-oxidative property.

Diesel exhaust particulate matter (DPM), which has been clarified as a Group I carcinogenic agent, is still challenging in its detoxification due to the complex composition and toxic mechanisms. Astaxanthin (AST) is a pleiotropic small biological molecule widely used in medical and healthcare with surprising effects and applications. The present study aimed to investigate the protective effects of AST on DPM-induced injury and the underlying mechanism. Our results indicated that AST significantly suppressed the generation of phosphorylated histone H2AX (γ-H2AX, marker of DNA damage) and inflammation caused by DPM both in vitro and in vivo. Mechanistically, AST prevented the endocytosis and intracellular accumulation of DPM via regulating the stability and fluidity of plasma membranes. Moreover, the oxidative stress elicited by DPM in cells could also be effectively inhibited by AST, together with protecting the structure and function of mitochondria. These investigations provided clear evidence that AST notably reduced DPM invasion and intracellular accumulation by modulating the membrane-endocytotic pathway, which eventually reduced intracellular oxidative stress caused by DPM. Our data might provide a novel clue for curing and treating the harmful effects of particulate matter.

Pulmonary flora-modified diesel particulate matter induced lung injury via cGAS signaling pathway.

Diesel particulate matter (DPM) is a major component of Fine Particulate Matter (PM2.5), which has been recognized by the World Health Organization under the name "Class I Carcinogen". Lung microbial communities are present widely in the lung tissue of a variety of organisms and play a significant role in the development and progression of lung disease, while cGAS is a DNA receptor that senses the invasion of microbial pathogens and activates the innate immune response. However, the role of cGAS in pulmonary flora-mediated PM2.5-induced lung injury is still largely unknown. With constructed cGAS-/- C57BL/6J mice, we found that lung damage, inflammation, and genetic damage induced by DPM were significantly blocked. With antibiotic-treated C57BL/6J mice, we found that healthy lung microbes were able to attenuate DPM-induced lung damage, inflammation, and genetic damage. DPM modified the expression of the cGAS/STING signaling pathway through the lung flora. This study revealed that cGAS signaling pathway played an essential role in lung flora-mediated adverse effects of DPM, which provided new therapeutic targets for lung diseases.

Multi-locus deletion mutation induced by silver nanoparticles: Role of lysosomal-autophagy dysfunction.

Due to the rapid production growth and a wide range of applications, safety concerns are being raised about the genotoxic properties of silver nanoparticles (AgNPs). In this research, we found AgNPs induced a size-dependent genotoxicity via lysosomal-autophagy dysfunction in human-hamster hybrid (AL) cells. Compared with 25 nm and 75 nm particles, 5 nm AgNPs could accentuate the genotoxic responses, including DNA double-strand breaks (DSBs) and multi-locus deletion mutation, which could be significantly enhanced by autophagy inhibitors 3-methyl adenine (3-MA), Bafilomycin A1 (BFA), and cathepsin inhibitors, respectively. The autophagy dysfunction was closely related to the accumulation of 5 nm AgNPs in the lysosomes and the interruption of lysosome-autophagosome fusion. With lysosomal protective agent 3-O-Methylsphingomyelin (3-O-M) and endocytosis inhibitor wortmannin, the reactivation of lysosomal function and the recovery of autophagy significantly attenuated AgNP-induced genotoxicity. Our data provide clear evidence to illustrate the role of subcellular targets in the genotoxicity of AgNPs in mammalian cells, which laid the basis for better understanding the health risk of AgNPs and their related products.

The Differential Antagonistic Ability of Curcumin against Cytotoxicity and Genotoxicity Induced by Distinct Heavy Metals.

Widespread heavy metal pollution has aroused severe health risks worldwide. Curcumin has been reported to play a wide-spectrum protective role for various heavy metals. However, the specificity and difference in the antagonistic ability of curcumin against distinct types of heavy metals are still largely unknown. Here, using cadmium (Cd), arsenic (As), lead (Pb), and nickel (Ni) as the typical heavy metals, we systematically compared the detoxification efficiency of curcumin on the cytotoxicity and genotoxicity elicited by different heavy metals under the same experimental conditions. Curcumin was proved to have a significant discrepant antagonistic capacity when counteracting the adverse effect of different heavy metals. Stronger protective effects of curcumin emerged when antagonizing the toxicity of Cd and As, rather than Pb and Ni. Curcumin exhibits a better detoxification ability against heavy metal-induced genotoxicity than cytotoxicity. Mechanistically, inhibiting the oxidative stress elicited by heavy metals and reducing the bioaccumulation of metal ions both contributed to the detoxification of curcumin against all the tested heavy metals. Our results illustrated that curcumin shows prominent detoxification specificity against different types of heavy metals and toxic endpoints, which provides a new clue for the better and targeted application of curcumin in heavy metal detoxification.

EPR-based in situ enzymatic activity detection of endogenous caspase-3 in apoptosis cell lysates.

Caspase-3 plays a vital role in cell apoptosis and related diseases. The detection and characterization of endogenous active caspase-3 are of immense value not only for mechanism studies of apoptosis but also for the diagnosis and treatment of apoptosis-related diseases. Here, an electron paramagnetic resonance (EPR)-based enzymatic assay was developed for the detection of caspase-3 activity both in vitro and in apoptosis cells. This assay uses a sandwich-like probe composed of a caspase-3-specific peptide segment (DEVD) conjugated to an EPR-detectable nitroxide spin label and magnetic beads (MBs). Cleavage of the "Nitroxide-Peptide-MBs" sandwich probe caspase-3 will release the nitroxide, which is readily detected by EPR after magnetic separation, resulting in a distinct EPR "off/on" transition. This assay takes advantage of the specific cleavage of DEVD-containing peptides by caspase-3 for high specificity, magnetic beads for fast magnetic separation, and EPR spectroscopy for considerably high detection sensitivity (LODs for caspase-3 are 116 nM at 60 min and 58 nM at 120 min). Importantly, the assay was proven to be compatible with complex biological samples and can detect the endogenous active caspase-3, thereby providing potential applications in the screening of protease-targeted drugs and the diagnosis of protease-associated diseases.

Zinc oxide/graphene oxide nanocomposites efficiently inhibited cadmium-induced hepatotoxicity via releasing Zn ions and up-regulating MRP1 expression.

Environmental cadmium (Cd) pollution has been verified to associated with various hepatic diseases, as Cd has been classified as one of the TOP 20 Hazardous Substances and liver is the main target of Cd poisoning. However, to design efficient hepatic antidotes with excellent detoxification capacity and reveal their underlying mechanism(s) are still challenges in Cd detoxification. Herein, ZnO/GO nanocomposites with favorable biocompatibility was uncovered their advanced function against Cd-elicited liver damage at the in situ level in vivo by 9.4 T magnetic resonance imaging (MRI). To explore the cellular detoxification mechanism, ZnO/GO nanocomposites was found to effectively inhibit the cyto- and geno-toxicity of Cd with the maximum antagonistic efficiency to be approximately 90%. Mechanistically, ZnO/GO nanocomposites competitively inhibited the cellular Cd uptake through releasing Zn ions, and significantly promoted Cd excretion via targeting the efflux pump of multidrug resistance associated protein1 (MRP1), which was confirmed by mass spectra and immunohistochemical analysis in kidney, a main excretion organ of Cd. Our data provided a novel approach against Cd-elicited hepatotoxic responses by constructed ZnO/GO nanocomposites both in vitro and in vivo, which may have promising application in prevention and detoxification for Cd poisoning.

Inokosterone Is A Potential Drug Target of Estrogen Receptor 1 in Rheumatoid Arthritis Patients: Analysis from Active Ingredient of Cyathula Officinalis.

OBJECTIVE: To elucidate the active compounds and the molecular mechanism of Cyathula Officinalis as a drug treatment for rheumatoid arthritis (RA). METHODS: The target genes of active ingredients from Cyathula Officinalis were obtained from bioinformatics analysis tool for the molecular mechanism of traditional Chinese medicine. The protein-protein interaction between the target genes were analyzed using STRING and Genemania. The transcriptome of RA patients compared to healthy people (GSE121894) were analyzed using R program package Limma. The relative expression of the target genes was obtained from the RNA-seq datasets. The molecular docking analyses were processed based on the molecular model of estrogen receptor 1 (ESR1) binding with estradiol (PDB ID:1A52). The binding details were analyzed by SYBYL. RESULTS: Inokosterone, ecdysterone, and cyaterone were the 3 active ingredients from Cyathula Officinalis that bind to target genes. Of all the significantly changed genes from RA patients, ESR1, ADORA1, and ANXA1 were significantly increased in mRNA samples of RA patients. CONCLUSION: ESR1, the transcription factor that binds inokosterone in the molecular binding analysis, is the target protein of Cyathula Officinalis.