Human & experimental toxicology最新文献

IntroductionHepatotoxicity can arise secondary to several medical conditions, including inflammation, sepsis, and therapy. Lipopolysaccharide (LPS) can activate diverse inflammatory pathways and has been implicated in hepatotoxicity. The store-operated calcium entry (SOCE), a key process for maintaining cellular calcium homeostasis, was shown to modulate inflammatory signaling. Reactive oxygen species, which serve a significant role in maintaining cellular function and homeostasis, are often elevated during inflammation, contributing to tissue injury. Therefore, we hypothesized that blocking the SOCE pathway would inhibit LPS-induced hepatotoxicity by suppressing inflammation and oxidative stress.MethodsTo test this, female BALB/c mice were randomly divided into the following experimental groups: control, LPS, LPS + SOCE inhibitor 2-aminoethoxy diphenyl borate (2APB), and 2APB alone. After 24 h of treatment, serum and liver samples were collected from the mice for histopathological, biochemical, and molecular analyses.ResultsInhibition of SOCE led to a decrease in the elevated liver function enzymes (ALT and AST) and protected the liver parenchymal cells as observed by histopathological assessment. Furthermore, blockade of SOCE significantly suppressed the level of il-1b, il-6, and cox2 genes in the liver tissues of mice treated with LPS. The expression of antioxidant genes (gsta1 and gpx1) was also significantly reduced by LPS treatment, while SOCE inhibition only restored the gpx1 expression. Additionally, treatment with 2APB attenuated LPS-induced oxidative stress in the liver of mice.ConclusionCollectively, our work demonstrated the critical involvement of SOCE in regulating inflammation and oxidative stress associated with LPS treatment, thereby reducing hepatotoxicity.

ObjectiveThe study aimed to explore the inhibitory effect of stevioside on colorectal cancer and its molecular mechanism.MethodsColorectal cancer cells were selected for functional testing, including the following groups: 0 μM stevioside, 1 μM stevioside, 2.5 μM stevioside and 5 μM stevioside. CCK-8 kit and EdU staining were employed to assess the cell viability. Cell apoptosis was deterred by flow cytometry. Western blot assay was utilized to detect the protein expressions of cleaved-caspase-3, Bax, Bcl-2, E-cadherin and Vimentin. The polarization of macrophage was evaluated through flow cytometry, western blot and immunofluorescence staining. The effect of stevioside on the proliferation of tumor tissue was detected by tumor formation and immunohistochemical staining in nude mice.ResultsStevioside exhibited a significant concentration-dependent inhibitory effect on the proliferation, migration, and invasion of colorectal cancer cells, while promoting apoptosis in vitro. Following stevioside treatment, there was a notable reduction in tumor volume and weight observed. Flow cytometry and immunohistochemical staining results showed that compared with control group, CD86+ cell ratio was increased in stevioside treatment group, while the CD206+ cell ratio was decreased in stevioside treatment group. RT-qPCR analysis revealed that, compared to the control group, stevioside treatment significantly reduced the mRNA expressions of Arg-1 and IL-10, while concomitantly increasing the mRNA expressions of IL-12 and TNF-α in a concentration-dependent manner.ConclusionStevioside possesses the ability to significantly hinder the proliferation of colorectal cancer cells and induce apoptosis, the mechanism of which may be closely related to the regulation of macrophage M1 polarization.

BackgroundHepatocellular carcinoma (LIHC), a prevalent liver cancer with a grim prognosis due to high recurrence rates, is under scrutiny for its association with zinc finger proteins (ZNFs) in tumorigenesis. This study aims to create a prognostic model for LIHC incorporating ZNF-related genes.MethodsBy analyzing TCGA data, we identified differentially expressed genes (DEGs) between normal and LIHC samples, focusing on ZNF-related genes through univariate Cox and LASSO Cox regression. A multivariate Cox regression model was built, categorizing LIHC patients into high- and low-ZNFRS groups based on ZNF-related risk scores. Model performance was evaluated using ROC curves, with a nomogram integrating clinical data and ZNFRS. Immune microenvironment, enrichment analysis, mutations, and drug responses in LIHC were also explored.ResultsA prognostic model utilizing 10 ZNF-related genes accurately predicted LIHC survival. The low-risk group exhibited enhanced immune cell infiltration, contrasting with cell cycle and DNA replication enrichment in the high-risk group, which also displayed increased mutation rates. Promising drug candidates like SNS-314 and Decitabine warrant further investigation in LIHC treatment.ConclusionThis study introduces impactful prognostic markers for LIHC management, emphasizing the significance of ZNFs in predicting patient outcomes and guiding treatment strategies.

ObjectiveTo explore the mechanisms by which microplastic toxicity leads to DNA damage in mouse spermatocytes.MethodsWe randomly divided GC-2 cells into a control group and a polystyrene microplastic (PS) group and then evaluated the DNA fragmentation index (DFI) in these cells via a comet assay. Whole-transcriptome sequencing was performed on the basis of DFI results. GO and KEGG enrichment analyses were based on the results of the entire transcriptome sequencing. At the same time, we also performed q-PCR validation on some significantly expressed genes and drew a toxicological network diagram on PS and mouse spermatocytes.ResultsComet assay results revealed that the intake of PS increased the DFI of mouse spermatocytes. Whole-transcriptome sequencing revealed that 61 circRNAs, 132 lncRNAs, 40 miRNAs, and 140 mRNAs were differentially expressed between the control and PS groups. GO and KEGG analyses revealed some notable enrichment in cellular components, molecular functions, biological processes, and gene expression pathways such as the defense response to viruses, the defense response to symbionts, the RIG-I-like receptor, the NOD-like receptor, and the calcium signaling pathways. Q-PCR and the network analysis revealed that PS affects the DFI of mouse spermatocytes mainly by influencing immune responses.ConclusionPS may damage the sperm DNA and increase the DFI by affecting cellular immunity-related pathways and redox pathways such as the RIG-I-like receptor and NOD-like receptor signaling pathways.

IntroductionBisphosphonates and denosumab are the most common antiresorptive drugs (ARs). ARs may cause medication-related osteonecrosis of the jaw (MRONJ) as a side effect, characterized by osteonecrosis and ulceration of the oral mucosa. The pathophysiology of MRONJ remains partly unclear and there is currently no consensus on its multietiological background. Immunomodulatory medications such as corticosteroids and antiestrogens may affect MRONJ onset.MethodsAn in vitro 3D cell culture model of gingival, keratinocytes, and fibroblasts were used to elucidate the pathogenesis of MRONJ. Cell cultures were exposed to ARs, followed by exposures to corticosteroids or antiestrogens. Morphology and proliferation were evaluated.ResultsAmong the ARs, alendronate caused the most negative cellular changes, while zoledronate only had a few effects. Denosumab caused more morphological cell atypia and proliferation than other ARs. The combined exposures of individual ARs with corticosteroids had some additional negative effects on gingival cells, whereas ARs with antiestrogen had few effects. The results are partly inconsistent, indicating that 3D cell culture experiments may not be the most suitable method for studying the effects of ARs.ConclusionThis study suggests that alendronate and denosumab affect gingival cell growth in a 3D cell culture model. These effects are smaller than reported in previous monolayer studies.

BackgroundAccurate and prompt assessment of malathion intoxication severity remains a significant clinical challenge, often hampered by reliance on single diagnostic markers. This exploratory case series investigated the combined utility of rapid butyrylcholinesterase (BChE) activity measurements and gas chromatography-tandem mass spectrometry (GC-MS/MS) for quantifying urinary malathion to enhance diagnostic precision.MethodsWe investigated three independent patients admitted with acute malathion intoxication. BChE activity was measured using both a point-of-care (POCT) device and a laboratory-based enzyme-multiplied immunoassay technique (EMIT). Urinary malathion was quantified using a validated GC-MS/MS method.ResultsMalathion exposure was confirmed in all patients via urinary analysis. Strong per-case positive correlations (r ranging from 0.905 to 0.996) were observed between the two BChE measurement methods, though Bland-Altman analysis revealed noteworthy discrepancies (mean bias of 10%, limits of agreement ranging from -20% to 40%). Critically, statistically significant inverse correlations (p < 0.05) were identified between urinary malathion concentrations and both BChE activity measurements, underscoring the dynamic relationship between exposure and enzymatic inhibition.ConclusionThese findings, derived from a small, exploratory case series, suggest the importance of an integrated diagnostic approach for malathion intoxication. This combined strategy may support improved assessment of severity and prognosis in individual cases, offering insights into the pesticide's systemic impact and elimination kinetics, especially when exposure details are unclear. While rapid BChE tests are valuable for initial screening, their interpretation should occur within this multi-marker framework. The generalizability of these findings is limited by the small sample size, and no formal power calculation was performed.

ObjectiveThe study aimed to investigate the role of luteolin in alleviating POF and its underlying molecular mechanisms.MethodsPOF model was established in rats by intraperitoneal injection of 100 mg/kg cyclophosphamide. Then, rats in the treatment group received intragastric administration of with luteolin at doses of 25 mg/kg, 50 mg/kg and 100 mg/kg, respectively. Rats in POF model group were administered the same volume of saline intragastrically. The concentrations of E2, FSH, AMH, P, LH, SOD and MDA in serum were quantified using ELISA kits. H&E and TUNEL staining were employed to assess pathological alterations and apoptosis. The cellular localizations of 4-HNE, 8-OHdG and NTY were detected by immunohistochemistry staining. The PharmMapper database and UbiBrowser prediction were used for the prediction of luteolin interaction with RNF8/HDAC2.ResultsLuteolin treatment significantly increased serum levels of estradiol (E2) (P < 0.01) and anti-Müllerian hormone (AMH) (P < 0.01) while reducing follicle-stimulating hormone (FSH) and luteinizing hormone (LH) levels (P < 0.05), restoring ovarian function. Additionally, luteolin could significantly improve ovarian tissue morphology and reduce apoptosis. ELISA kit results indicated luteolin significantly reduced the levels of SOD and MDA. Immunohistochemical staining results revealed a significant decrease in the expressions of 4-HNE, 8-OHdG and NTY in luteolin treatment group. Combining The PharmMapper database and UbiBrowser prediction, we found luteolin could bind RNF8 and inhibit HDAC2 expression.DiscussionLuteolin could mitigate cyclophosphamide-induced ovarian senescence, with its molecular mechanisms involving the regulation of RNF8/HDAC2 signaling axis and the inhibition of oxidative stress.

BackgroundLipopolysaccharide (LPS) is a glycolipid that constitutes the Gram-negative bacteria outermost membrane main portion. LPS is frequently of concern in medicine because of nearly all severe sepsis patients have elevated LPS plasma levels, which cause life-threatening organ dysfunction. Consequently, the potential protective benefit of acriflavine (Ac) in limiting LPS-induced acute inflammatory response and the possible underlying mechanisms were investigated.MethodsMale albino mice were treated with i.p. Ac 4 or 8 mg/kg/day for 2 weeks, then received a single i.p. LPS (10 mg/kg) at day 14.ResultsAc administration ameliorated hepatic, pulmonary, and testicular dysfunction, as confirmed by attenuation of pathological changes and amendment of oxidative stress parameters. This was associated with inhibition of protein kinase R-like endoplasmic reticulum kinase/phosphatidylinositol 3-kinase (PERK/PI3K) endoplasmic reticulum (ER) stress pathway; toll-like receptor 4/nuclear factor kappa B (TLR4/NF-κB) and active caspase-1/gasdermin D (GSDMD)-N-terminal as well as interleukin-1 beta (IL-1β) inflammatory and pyroptotic signals.ConclusionOur results highlighted the protective potential of Ac in a mouse model of LPS-mediated systemic inflammatory response, which paves the way for its clinical application in sepsis.

IntroductionThis study investigates the skin-irritating and skin-resorptive effects of drilling fluid (DF) through acute and subacute experiments conducted on adult rabbits and sexually mature male and female rats.MethodsAcute and subacute experiments were conducted on adult rabbits and sexually mature male and female rats. The acute experiment involved a single exposure to DF, while the subacute study involved repeated dermal exposure. Various physiological, biochemical, and histopathological assessments were performed to evaluate the effects.ResultsThe results of the acute experiment demonstrated that DF exhibits a mild skin-irritating effect but causes significant irritation to the ocular mucous membranes in rabbits. In the subacute study, dermal exposure to DF led to notable alterations in the physiological and biochemical status of rats, including reduced food and water intake, decreased body weight gain, and significant changes in hematological and biochemical parameters. An increase in the relative count of certain leukocyte populations was observed, with a statistically significant elevation in absolute eosinophil counts in both sexes. Similar trends were observed for basophils and lymphocytes. Moderate elevations in serum enzyme levels (ALT, AST, ALP, GGT, and LDH) indicated systemic toxicity. Morphological and morphometric analysis of the skin further supported the dermal toxicity of DF, revealing epidermal keratinization, marked proliferation of germinative layer cells, and cellular heterotopia. In the basal layer of the epidermis, cells exhibited signs of intracellular edema. The papillary layer showed moderate focal inflammatory infiltrates, while the reticular dermis displayed edema of fibrous structures, swelling of collagen fibers, and increased fiber thickness.DiscussionThese findings collectively demonstrate that the tested drilling fluid possesses significant dermal and systemic toxicity, indicating potential health risks for mammals upon repeated exposure.