Molecular & Cellular Toxicology最新文献

Background

Benzene is a toxic pollutant in industrial production that can cause serious damage to haematopoietic system, and the liver is the most important organ for benzene metabolism.

Objective

The aim of this study was to explore reliable biomarkers of benzene poisoning for an in-depth study of liver damage and its possible metabolic mechanisms through 4D label-free proteomics. We established a chronic benzene poisoning model in C57BL/6J mice via the gavage of a benzene/peanut oil mixture. Micebody weights and routine blood test results were recorded in detail. Mice livers were collected, and 4D label-free proteomics was used to identify the differentially expressed proteins.

Results

Compared with the control group, slow body weight gain, a reduction in whole blood cells and hepatocyte oedema were observed in the benzene poisoning group. A total of 303 differentially expressed proteins were identified with the screening conditions of a fold change > 2 (or < 1/2) and a P value < 0.05, of which 127 proteins were significantly upregulated and 176 proteins were significantly downregulated. Kyoto Encyclopedia of Genes and Genomes analysis revealed that these proteins were associated mainly with metabolic pathways, metabolism of xenobiotics by cytochrome P450 and steroid hormone biosynthesis. We further selected three core proteins, Cytochrome P450 2B10, NADH dehydrogenase [quinone] 1 and Glutathione S-transferase Mu 3, for dual validation via immunoblotting and immunohistochemistry.

Conclusion

This study contributes to our understanding of benzene-induced hepatotoxicity and its metabolic mechanism in the liver.

Background

High salt and hyperosmolarity can cause renal cell death, which can act as a risk factor for a variety of conditions, including acute and chronic kidney disease. Therefore, a monitoring system to assess renal cytotoxicity is required.

Objectives

The study aimed to develop a system that could rapidly and accurately assess the properties of stress granules, non-membrane organelles that are regulated by liquid–liquid phase separation and form when cells are stressed, by exploiting the properties of stress granules (SGs).

Results

We established a human embryonic kidney cell line expressing endogenous Ras GTPase-activating protein-binding protein 1 (G3BP1)-green fluorescent protein (GFP) through CRISPR/Cas9 gene editing. We found that G3BP1-GFP cells formed SGs similar to native G3BP1 cells after exposure to arsenite, high salt, and osmotic stress. We also validated the human embryonic kidney cell line expressing G3BP1-GFP through real-time monitoring.

Conclusion

The G3BP1-GFP expressing human embryonic kidney cell line provides a novel method to assess kidney toxicity through real-time monitoring of SGs. This method allows for real-time monitoring of SGs in response to various renal toxicants, providing a sensitive and rapid approach for toxicity assessment.

Background

Parkinson’s disease (PD) is a common neurodegenerative disease in middle-aged and older adults. Autophagy defect is related to the pathogenesis of Parkinson’s disease. Ursolic acid (UA), a naturally occurring triterpene carboxylic acid compound, has recently gained attention for its strong antioxidant properties. However, little is known about the effect of UA on the level of apoptosis of nerve cells.

Objective

This study aims to investigate whether UA could regulate autophagy-dependent apoptosis by reducing the level of cellular oxidative stress.

Results

Here, we found that Rotenone led to a reduction in ATG5/LC3-II levels and autophagosome formation in PC12 cells and primary neurons, accompanied by an increase in p62 level, indicating inhibition of autophagy. However, interestingly, knocking down ATG5 enhanced the inhibition of autophagy and increased neuronal apoptosis by Rotenone. The effects of Rotenone on autophagy and apoptosis were related to the enhancement of oxidative stress and the activation of AMP-activated protein kinase (AMPK). UA decreased the oxidative stress level of nerve cells, inhibited the activity of AMPK, increased the autophagosome formation and ATG5/LC3-II levels, and decreased the p62 level, promoting autophagy and thereby suppressing apoptosis. Furthermore, dominant-negative AMPKα, or inhibition of AMPK with compound C alleviated Rotenone-induced decrease in ATG5/LC3-II levels and autophagosome formation, and increases in phosphorylated AMPK and p62 levels, and cell apoptosis. The effect of Rotenone on autophagy and apoptosis is associated with the production of excess oxygen radicals in cells, as evidenced using UA.

Conclusion

These results suggest that UA can alleviate AMPK activation, inhibition of autophagy, and neuronal apoptosis induced by Rotenone by reducing oxidative stress level. Our findings highlight that UA has great potential in the prevention and treatment of Parkinson’s disease.

Background

Salvia microphylla (S. microphylla), a plant grown as a medicinal plant in central Mexico and used to make tea, has been demonstrated to possess anticancer, anti-inflammatory, and antifungal properties. However, the anticancer properties of S. microphylla essential oil have not been investigated. Hence, we conducted this experiment to highlight its efficacy against cancer.

Methods

Three different types of ovarian cancer cells were incubated with S. microphylla essential oil concentration dependently and detected at 560 nm and 650 nm of microplate reader. The specific components of extract were detected by GC–MS. Cell apoptosis, cytosolic and mitochondrial calcium level, reactive oxygen species, and mitochondrial membrane potential were detected by flow cytometry. The mRNA expressions of apoptosis, antioxidants, and mitochondria-related genes were measured by real-time qPCR.

Results

S. microphylla essential oil inhibited the viability of a cisplatin-resistant ovarian cancer cell line A2780cis, with intracellular calcium imbalance, reactive oxygen species regulation, mitochondria membrane potential loss and cell cycle arrest. Moreover, expressions of genes affecting mitochondrial complex, mitochondrial membrane pores, antioxidant levels, endoplasmic reticulum stress, and apoptosis were investigated.

Conclusion

The S. microphylla essential oil has anticancer potential in A2780cis, encouraging further in vivo studies.

Background

M2-polarized macrophages aggressively modulate the tumor microenvironment and enhance tumor cell malignancy. As intracellular molecules are released by damaged or stressed cells, damage-associated molecular patterns (DAMPs) bind to toll-like receptors (TLRs) on cells in the tumor microenvironment, inducing inflammation and epithelial-mesenchymal transition. However, recent studies on the crosstalk between DAMPs and M2-polarized macrophages (M2 macrophages) during tumor progression have not provided conclusive results.

Objective

We investigated the role of toll-like receptors (TLRs) in IL-6 production by M2 macrophages and searched for cancer cell-derived DAMPs that can activate the TLRs responsible for IL-6 production.

Results

TLR2 activation was required for IL-6 production by M2 macrophages. The malignancy of cancer cells was increased by the activation of this pathway. Cancer-derived HSP 72 acted as a ligand that stimulates the TLR2 signaling pathway in M2 macrophages, triggering IL-6 production.

Conclusion

TLR 2 stimulation in M2 macrophages enhances tumor malignancy by upregulating IL-6. Heat shock protein 72 (HSP72) is a potent TLR2 stimulator. Our findings reveal a connection between TLR2 and M2-polarized macrophages in tumor malignancy and may be useful for developing effective treatments for tumor relapse.

Background

Akebia quinata D., a plant from the Lardizabalaceae family, has been traditionally used in East Asia for its medicinal properties. Its dried stem serves as a diuretic agent for treating edema and rheumatic pain.

Objective

This study aimed to explore the anti-inflammatory activity of A. quinata D. stem extracts in lipopolysaccharide (LPS)-induced RAW 264.7 macrophages. We also investigated potential mechanisms involving the mitogen-activated protein kinases (MAPK) and NF-kB signaling pathways.

Results

Extracts were obtained from dried A. quinata D. stems using ethanol (AQDE) and water (AQDW) extraction methods. The effects of A. quinata D. extracts on cytotoxicity, anti-inflammatory activity in RAW 264.7 macrophages were analyzed. A. quinata D. extracts were non-toxic in various concentrations (25–800 μg/mL) in RAW 264.7 cells. Treatment with extracts significantly reduced nitric oxide production, PGE2, proinflammatory cytokines (TNF-α, IL-1β, and IL-6), and PTGES2 in LPS-induced RAW 264.7 cells. Downregulation of the MAPK signaling pathway led to reduced phosphorylation of NF-κB, a transcription factor modulating inflammatory proteins, including JNK, ERK1/2, and p38. Based on the experimental results, it was confirmed that AQDE exhibits superior anti-inflammatory effects than AQDW.

Conclusion

These results indicate that A. quinata D. extracts exhibit anti-inflammatory effects without toxicity. Therefore, this suggests that they hold promise as a therapeutic candidate for inflammatory diseases, as well as potential a functional material for cosmeceuticals, food and medicine industries.

Background

Cancer stem cells play important roles in tumorigenesis and progression of glioma. Circular RNA nuclear factor I X (circNFIX, hsa_circ_0049658) has been found to facilitate the development of glioma. The regulation of circNFIX in glioma stem cells (GSCs) has never been studied.

Methods

RNA levels of circNFIX, microRNA-449a (miR-449a) and cluster of differentiation 47 (CD47) were detected through reverse transcription-quantitative polymerase chain reaction (RT-qPCR). The protein analysis was performed by western blot. Cell proliferation and self-renewal abilities were assessed by EdU assay and tumor sphere formation assay. The macrophage-mediated phagocytosis and cytotoxicity were examined by flow cytometry and lactate dehydrogenase (LDH) assay, respectively. The target analysis was performed using dual-luciferase reporter assay, RNA immunoprecipitation (RIP) and RNA pull-down assays. In vivo assay was conducted through GSCs model in mice.

Results

The levels of circNFIX and CD47 were upregulated in glioma tissues and GSCs. Silencing circNFIX or CD47 suppressed proliferation and self-renewal abilities but enhanced phagocytosis and cytotoxicity in GSCs. CircNFIX induced the positive regulation of CD47 by exerting the sponge effect on miR-449a in GSCs. The regulatory role of circNFIX was achieved by targeting miR-449a in GSCs. CircNFIX downregulation also repressed tumorigenesis of glioma and maintenance of stem cells by the miR-449a/CD47 axis in GSCs model in vivo.

Conclusion

The evidence suggested that circNFIX acted as a miR-449a sponge to regulate the CD47 level, thereby promoting the features of glioma stem cells and the evasion of phagocytosis.

Background

Atopic dermatitis (AD) is a chronic inflammatory disorder that can affect the physical and psychological health of individuals worldwide. Mangiferin has been previously shown to alleviate allergic diseases.

Objectives

To investigate the effect of mangiferin on AD-like pathologies and the mechanism of action.

Methods

After establishing a mouse model of AD by challenging male BALB/c mice with 1% 2,4-dinitrochlorobenzene (DNCB), 10, 50, and 100 mg/kg mangiferin was administered orally for 21 days. Scratching behaviors, dermatitis score, spleen weight, and serum immunoglobulin E (IgE) levels were assessed. Histopathological changes were determined using toluidine blue and hematoxylin–eosin staining. A cellular model of AD was established by co-stimulating HaCaT keratinocytes with tumor necrosis factor (TNF)-α and interferon (IFN)-γ. The mRNA expression of chemokines and Th2-related cytokines was examined using RT-qPCR. Western blot and immunofluorescence were used to detect the protein levels of markers of the AKT/NF-κB/STAT1 and MAPK pathways.

Results

Mangiferin exhibited anti-atopic effects in DNCB-induced AD-like mice as evidenced by decreased scratching behaviors, dermatitis score, spleen weight, IgE levels, skin thickness, and mast cell infiltration in mice. Additionally, mangiferin inhibited expression of chemokines and Th2 type cytokines (IL-4, IL-5, and IL-13) in AD-like mice and TNF-α/IFN-γ-co-stimulated HaCaT cells. Moreover, mangiferin attenuated TNF-α/IFN-γ-induced release of various inflammatory factors (such as TNF-α, IL-1β, and IL-6) in HaCaT cells. Mangiferin repressed the activation of the AKT/NF-κB/STAT1 and MAPK signaling pathways under both in vivo and in vitro conditions.

Conclusion

Overall, mangiferin inhibits the inflammatory and pruritic responses in AD, which might be an effective drug for AD treatment.

Background

Perfluorooctanoic acid (PFOA) and its alternative compound, hexafluoropropylene oxide dimer acid (HFPO-DA), are commonly used in cookware and food packaging because they are structurally stable and have a long half-life. This allows them to enter the human body, where they accumulate and have potentially harmful effects on human health. While the ability of PFOA to induce various cytotoxicities, including endoplasmic reticulum (ER) stress, has been studied extensively, little is known about the effects of alternative compounds, such as HFPO-DA, on cells.

Objectives

This study aimed to evaluate the gastrointestinal toxicity of PFOA and HFPO-DA in vitro using a gastrointestinal AGS cell line. We established a real-time stress granule reporter system in human gastrointestinal cell lines by inserting green fluorescent protein into the Ras-GTPase-activating protein SH3-domain-binding protein 1 gene using CRISPR/Cas9-mediated homologous recombination.

Results

We found that PFOA induced the phosphorylation of PERK, which is activated by ER stress in AGS cells, and in contrast, the alternative compound HFPO-DA did not induce the phosphorylation of PKR-like endoplasmic reticulum kinase under the same conditions as PFOA.

Conclusion

Based on our results, we concluded that, unlike PFOA, HFPO-DA does not induce ER stress, suggesting that it is less toxic than PFOA. However, the risks of HFPO-DA cannot be dismissed completely, and further studies comprising different stress conditions are needed to accurately predict the potential risks of alternative compounds, such as HFPO-DA.

Background

Renal ischemia/reperfusion injury (RI/RI) is the crucial cause of acute kidney injury.

Objective

The study was to explore the action mechanism of desflurane (DFE) pretreatment on RI/RI in rats. The RI/RI model of rats was constructed, and serum renal function parameters (blood urea nitrogen and serum creatinine) were detected adopting corresponding commercial kits to assess renal function injury. Detection of renal tissue damage was performed. Test of cell apoptosis was implemented. Examination of endoplasmic reticulum stress (ERS)-associated proteins C/EBP homologous protein, glucose regulatory protein 78, and activation transcription factor 6 was performed. Test of concentrations of interleukin-1β and tumor necrosis factor-αwas implemented.

Result

The results illustrated DFE restrained renal ischemia–reperfusion (RIR)-stimulated ERS and apoptosis, and DFE elevated miR-181c-5p and suppressed DDIT4 inRIR rats. MicroRNA (miR)-181c-5p was declined in RIR rats and ameliorated ischemia–reperfusion-stimulated ERS and apoptosis, suppressive miR-181c-5p or elevated DDIT4 turned around the protection of DFE on RIR. MiR-181c-5p targeted DNA damage-inducible transcript 4 (DDIT4).

Conclusion

The study offered theoretical foundation for further exploration on DFE in repressing RIR-stimulated ERS and apoptosis via elevating miR-181c-5p to target DDIT4.