ABSTRACT
Microplastics are detrimental to the environment. However, the combined effects of microplastics and arsenic (As) remain unclear. In this study, we investigated the combined effects of polystyrene (PS) microplastics and As on HepG2 cells. The results showed that PS microplastics 20, 50, 200, and 500 nm in size were taken up by HepG2 cells, causing a decrease in cellular mitochondrial membrane potential. The results of lactate dehydrogenase release and flow cytometry showed that PS microplastics, especially those of 50 nm, enhanced As-induced apoptosis. In addition, transcriptome analysis revealed that TP53, AKT1, CASP3, ACTB, BCL2L1, CASP8, XIAP, MCL1, NFKBIA, and CASP7 were the top 10 hub genes for PS that enhanced the role of As in HepG2 cell apoptosis. Our results suggest that nano-PS enhances As-induced apoptosis. Furthermore, this study is important for a better understanding of the role of microplastics in As-induced hepatotoxicity.
Subject(s)
Arsenic , Humans , Arsenic/toxicity , Hep G2 Cells , Microplastics/toxicity , Plastics , Polystyrenes/toxicity , ApoptosisABSTRACT
In this study, we investigated the effect of caffeine overexposure on corneal innervation in the early chicken embryo. Caffeine administration restricted corneal innervation by affecting trigeminal nerve development. Immunohistochemistry for phospho-Histone3 (pHIS3) and C-caspase3 revealed that cell survival was repressed by caffeine administration. Whole-mount in situ hybridization against semaphorin 3A (Sema3A) and neuropilin-1 (Nrp1) showed that both caffeine and 2,2'-azobis(2-methylpropionamidine) dihydrochloride (AAPH, a free radical generator) administration upregulates the expression of both Sema3A and Nrp1. Next, we demonstrated that lens ablation in the developing chicken embryos significantly affected NF-labeled periocular nerve fascicles and innervation to the central eye region. Subsequently, we used a neuroblastoma cell line to investigate in vitro whether or not Sema3A-Nrp1 signaling exerts a key role on the caffeine-suppressed neuron survival. Knocking-down Sema3A through transfection with Sema3A-siRNA dramatically decreased the responsiveness of cells to caffeine administration, as well as cell apoptosis. We suggest that Sema3A-Nrp1 signaling regulates Trp53 and Cdkn1a through Slit2-Robo1 and Ephb2. Taken together, we speculate here that caffeine-enhanced reactive oxygen species upregulates Sema3A-Nrp1 expression in the lens and periocular tissues, resulting in corneal cell apoptosis, accompanied by its chemorepellent role on the invasion of the developing cornea by trigeminal sensory fibers.
Subject(s)
Neuropilin-1/metabolism , Semaphorin-3A/metabolism , Animals , Caffeine/pharmacology , Cell Line, Tumor , Chick Embryo , Cornea , Gene Expression Regulation, Developmental/drug effects , Gene Expression Regulation, Developmental/physiology , Gene Knockdown Techniques , Humans , Lens, Crystalline , Nerve Tissue Proteins/genetics , Nerve Tissue Proteins/metabolism , Neuroblastoma , Organogenesis , Reactive Oxygen Species , Receptors, Immunologic/genetics , Receptors, Immunologic/metabolism , Roundabout ProteinsABSTRACT
The enrichment of various pollutants in mangrove has attracted widespread attention. Especially, microplastics accumulation in mangrove may provide a more challenging ecological colonization site by enriching pollutants, thus affecting the change of microplastics antibiotic resistance and increasing the risk of antibiotic failure. Herein, the antibiotic-resistant of microplastics and sediment from mangrove were investigated. The results show that isolates are mainly colonized by Vibrio parahemolyticus (V. parahemolyticus), Vibrio alginolyticus (V. alginolyticus), and Shewanella. 100% mangrove microplastics isolates are resistant to chloramphenicol, cefazolin, and tetracycline, especially amoxicillin clavulanate and ampicillin. Meanwhile, the multiple antibiotics resistance (MAR) indexes of V. parahaemolyticus, Shewanella, and V. alginolyticus in mangrove microplastics are 0.72, 0.77, and 0.77, respectively, which are far higher than the MAR index standard (0.2) and that of mangrove sediment isolates. Furthermore, compared with V. parahaemolyticus isolated from the same mangrove microplastics, Shewanella and V. alginolyticus show stronger drug resistance. It should be noted that there is a closely related relationship between the type of microplastics and the antibiotics resistance of isolated bacteria. For the antibiotics sensitivity test of norfloxacin, streptomycin, amoxicillin, and chloramphenicol, V. parahaemolyticus have the lower antibiotics resistance than that of V. alginolyticus isolated from the same mangrove microplastics. However, Vibrio isolated from PE has stronger antibiotics resistance. Results reveal that mangrove may be one of the potential risks for emergence and spread of bacterial antibiotics-resistant and multidrug-resistant, and microplastic biofilms may act as promoters of bacterial antibiotic resistance.