Journal of Toxicological Sciences最新文献

{"title":"Effects of microsampling on toxicity evaluation of 1-naphthylisothiocyanate (ANIT), a hepatotoxic substance, in a mouse toxicity study.","authors":"Kazuaki Takahashi,&nbsp;Shun Kawaguchi,&nbsp;Tomohiro Ikeda,&nbsp;Yuki Tomonari,&nbsp;Takeshi Funakoshi,&nbsp;Keiko Nakai,&nbsp;Tooru Fujimoto,&nbsp;Dai Yamamoto,&nbsp;Takayuki Okamura,&nbsp;Hideomi Uchida,&nbsp;Yoshiro Saito,&nbsp;Seiji Otake","doi":"10.2131/jts.48.607","DOIUrl":"https://doi.org/10.2131/jts.48.607","url":null,"abstract":"<p><p>ICH S3A Q&A focused on microsampling (MS) was published to help accelerate the use of MS and states that MS is useful because toxicokinetic (TK) evaluation with conventional blood sampling volume requires many animals for TK satellite groups; however, there are few reports of MS application in mice. We investigated the influence of MS on toxicity evaluation in mice by comparing the toxicity parameters with and without MS after a single oral administration of 1-naphthylisothiocyanate (ANIT), a hepatotoxic substance. Blood samples (50 µL/point) were collected from the tail vein of 3 mice per group at 2 or 3 time points during a 24-hr period, and toxicity was evaluated 2 days after administration. ANIT-related changes suggesting liver or gallbladder injury were noted in blood chemistry and histopathology. Some of these changes such as increases in focal hepatocyte necrosis and inflammatory cell infiltration in the liver as well as mucosal epithelium necrosis in the gallbladder were apparently influenced by MS. A tendency to anemia was noted in animals with MS but not without MS, which was also noted in the vehicle-treated controls, suggesting influence of blood loss. The current results indicate that ANIT hepatotoxicity could be evaluated in mice in which blood samples were collected by MS for most parameters; however, parameters in anemia and pathology in the liver and gallbladder were influenced by MS in this study condition with ANIT. Therefore, MS application in mice should be carefully considered.</p>","PeriodicalId":17654,"journal":{"name":"Journal of Toxicological Sciences","volume":"48 11","pages":"607-615"},"PeriodicalIF":2.0,"publicationDate":"2023-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"71424743","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Pathogenesis of selective damage of granule cell layer in cerebellum of rats exposed to methylmercury.","authors":"Ke Du,&nbsp;Takashi Hirooka,&nbsp;Yu Sasaki,&nbsp;Akira Yasutake,&nbsp;Takato Hara,&nbsp;Chika Yamamoto,&nbsp;Yasuyuki Fujiwara,&nbsp;Yo Shinoda,&nbsp;Tomoya Fujie,&nbsp;Shogo Katsuda,&nbsp;Komyo Eto,&nbsp;Toshiyuki Kaji","doi":"10.2131/jts.48.429","DOIUrl":"https://doi.org/10.2131/jts.48.429","url":null,"abstract":"<p><p>Granule cell-selective toxicity of methylmercury in the cerebellum is one of the main unresolved issues in the pathogenesis of Minamata disease. Rats were orally administered methylmercury chloride (10 mg/kg/day) for 5 consecutive days, and their brains were harvested on days 1, 7, 14, 21, or 28 after the last administration for histological examination of the cerebellum. It was found that methylmercury caused a marked degenerative change to the granule cell layers but not to the Purkinje cell layers. The generative change of the granule cell layer was due to cell death, including apoptosis, which occurred at day 21 and beyond after the methylmercury administration. Meanwhile, cytotoxic T-lymphocytes and macrophages had infiltrated the granule cell layer. Additionally, granule cells are shown to be a cell type susceptible to TNF-α. Taken together, these results suggest that methylmercury causes small-scale damage to granule cells, triggering the infiltration of cytotoxic T-lymphocytes and macrophages into the granule cell layer, which secrete tumor necrosis factor-α (TNF-α) to induce apoptosis in granule cells. This chain is established based on the susceptibility of granule cells to methylmercury, the ability of cytotoxic T lymphocytes and macrophages to synthesize and secrete TNF-α, and the sensitivity of granule cells to TNF-α and methylmercury. We propose to call the pathology of methylmercury-induced cerebellar damage the \"inflammation hypothesis.\"</p>","PeriodicalId":17654,"journal":{"name":"Journal of Toxicological Sciences","volume":"48 7","pages":"429-439"},"PeriodicalIF":2.0,"publicationDate":"2023-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"10121230","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Plasma membrane damage triggered by benzalkonium chloride and cetylpyridinium chloride induces G₀/G₁ cell cycle arrest via Cdc6 reduction in human lung epithelial cells.","authors":"Sanae Kanno,&nbsp;Seishiro Hirano,&nbsp;Jun Monma-Otaki,&nbsp;Hideaki Kato,&nbsp;Mamiko Fukuta,&nbsp;Yoshimi Nakamura,&nbsp;Yasuhiro Aoki","doi":"10.2131/jts.48.75","DOIUrl":"https://doi.org/10.2131/jts.48.75","url":null,"abstract":"<p><p>Quaternary ammonium compounds, including benzalkonium chloride (BAC) and cetylpyridinium chloride (CPC), are widely used as disinfectants. Increased use of inhalable products containing BAC or CPC has raised concerns for lung toxicity. This study sought to elucidate the microstructure of plasma membrane damage caused by BAC and CPC and the subsequent mechanism by which the damage is mediated, as assessed using two human pulmonary epithelial cell lines (A549 and BEAS-2B). Scanning electron microscopic observation showed that exposure to BAC or CPC for 3 hr reduced the length and density of microvilli on the plasma membrane in A549 cells. Analysis of cell cycle distribution following plasma membrane damage revealed that BAC and CPC promote G₀/G₁ cell cycle arrest in both cell lines. The protein levels of Cdc6, an essential regulator of DNA replication during G₁/S transition, are decreased significantly and dose dependently by BAC or CPC exposure. CPC and BAC decreased the Cdc6 levels that had been increased by a PI3K agonist in A549 cells, and levels of phosphorylated AKT were reduced in response to BAC or CPC. Conversely, exposure to equivalent concentrations of pyridinium chloride (lacking a hydrocarbon tail) induce no changes. These results suggest that plasma membrane damage triggered by BAC or CPC causes Cdc6-dependent G₀/G₁ cell cycle arrest in pulmonary cells. These effects are attributable to the long alkyl chains of BAC and CPC. The reduction of Cdc6 following plasma membrane damage may be caused, at least in part, by diminished signaling via the PI3K/AKT pathway.</p>","PeriodicalId":17654,"journal":{"name":"Journal of Toxicological Sciences","volume":"48 2","pages":"75-86"},"PeriodicalIF":2.0,"publicationDate":"2023-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"9208510","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Establishment of hyperoxic cell culture system for predicting drug-induced liver injury: reducing accumulated lipids in hepatocytes derived from chimeric mice with humanized liver.","authors":"Yuya Ohtsuki,&nbsp;Seigo Sanoh,&nbsp;Mikaru Yamao,&nbsp;Yuha Kojima,&nbsp;Yaichiro Kotake,&nbsp;Chise Tateno","doi":"10.2131/jts.48.99","DOIUrl":"https://doi.org/10.2131/jts.48.99","url":null,"abstract":"<p><p>Drug-induced liver injury (DILI) is a major adverse reaction. Species-specific differences between humans and laboratory animals make it difficult to establish evaluation models that can accurately predict DILI in the preclinical phase. Chimeric mice with humanized liver are potential predictive models for understanding DILI. Chimeric mice generated by transplanting human hepatocytes into urokinase-type plasminogen activator/severe combined immunodeficient mice are known to develop fatty liver and show lipid accumulation in isolated hepatocytes. It is speculated that the lipids accumulated in hepatocytes may interfere with DILI assessment. It is known that normal 20% oxygen culture conditions do not meet oxygen demand because oxygen consumption rate is higher than the oxygen supply rate. Therefore, we predicted that hyperoxic cultures could induce hepatocyte function and reduce accumulated lipids. A culture of chimeric mouse hepatocytes in 40% oxygen showed reduced intracellular lipid and triglyceride levels compared to those cultured in 20% oxygen on days 7 and 10. In addition, fatty acid β-oxidation (FAO) activity increased from day 7 under 40% oxygen conditions. On the other hand, FAO activity increased on day 10 under 20% conditions. Microarray and Ingenuity Pathway Analysis showed that lipid metabolism-related pathways were downregulated under 40% oxygen conditions for 7 days, suggesting the involvement of several mechanisms in decreasing lipid levels and increasing FAO. Furthermore, some pathways related to cellular function and maintenance were upregulated under 40% oxygen conditions for 7 days. In conclusion, chimeric mouse hepatocytes cultured under hyperoxic conditions may be useful for predicting DILI.</p>","PeriodicalId":17654,"journal":{"name":"Journal of Toxicological Sciences","volume":"48 2","pages":"99-108"},"PeriodicalIF":2.0,"publicationDate":"2023-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"9208511","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Experience in the treatment of chlorfenapyr poisoning.","authors":"Benhe Wu,&nbsp;Fei Xue,&nbsp;Mingfeng Lu,&nbsp;Aiwen Ma,&nbsp;Lu Fan","doi":"10.2131/jts.48.221","DOIUrl":"https://doi.org/10.2131/jts.48.221","url":null,"abstract":"<p><p>In China, the extensive use of the pesticide chlorfenapyr has led to an increase in chlorfenapyr poisoning. However, there are limited reports on chlorfenapyr poisoning, and most of them are fatal cases. This study retrospectively analyzed four patients admitted to the emergency room after chlorfenapyr intake and detected different concentrations of chlorfenapyr in their plasma. Among them, one patient died and three patients survived. Case 1 suffered respiratory and circulatory failure with a deep coma shortly after oral administration of 100 mL of a the chlorfenapyr-containing mixture and died 30 min after admission. Case 2 experienced transient nausea and vomiting after oral administration of chlorfenapyr (50 mL). The patient had normal laboratory results and was discharged with no further treatment. Case 3 developed nausea and vomiting and a light coma after taking 30 mL of chlorfenapyr orally. He underwent blood perfusion and plasma exchange in the intensive care unit (ICU) and was discharged with recovery. A two-week follow-up visit, however, revealed hyperhidrosis. Case 4 (advanced age with severe underlying disease) developed a light coma after oral intake of 30 mL of chlorfenapyr. Subsequently, pulmonary infection and gastrointestinal bleeding were developed. The patient experienced blood perfusion and mechanical ventilation in the ICU and finally survived after treatment. The present study provides the basic information, plasma concentration of toxins, onset of poisoning and treatment process of the four patients mentioned above, providing novel insights into the clinical diagnosis and treatment of chlorfenapyr poisoning.</p>","PeriodicalId":17654,"journal":{"name":"Journal of Toxicological Sciences","volume":"48 4","pages":"221-225"},"PeriodicalIF":2.0,"publicationDate":"2023-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"9294540","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Oxidative stress and cellular toxicity induced by dihydropyrazine: a comparative study with other Maillard reaction products.","authors":"Yuu Miyauchi,&nbsp;Haruna Koba,&nbsp;Madoka Sawai,&nbsp;Hisao Kansui,&nbsp;Shinji Takechi","doi":"10.2131/jts.48.263","DOIUrl":"https://doi.org/10.2131/jts.48.263","url":null,"abstract":"<p><p>Glycation products are generated during the Maillard reaction, a non-enzymatic reaction between reducing sugars and the amino groups of proteins, which accumulate in the body with aging and cause many diseases. Herein, we have focused on dihydropyrazines (DHPs), which are glycation products formed by the dimerization of D-glucosamine or 5-aminolevulinic acid, and have reported that DHPs can produce several kinds of radicals and induce cytotoxicity via oxidative stress. To advance our understanding of DHP-mediated cytotoxicity, we selected a DHP, 3-hydro-2,2,5,6-tetramethylpyrazine (DHP-3), and two major Maillard reaction products, N^ε-(carboxymethyl)-L-lysine (CML) and acrylamide, and performed comparative experiments focusing on their cytotoxicity and their ability to induce oxidative stress. The order of increasing cytotoxicity was DHP-3, acrylamide, and CML, and the LC₅₀ value could be calculated only for DHP-3 (0.53 mM), indicating that DHP-3 is more toxic than the other Maillard reaction products. However, their toxicities were significantly lower than those of common toxic chemicals. Further, the results of their cytotoxicity assay were consistent with the results of intracellular reactive oxygen species production and activation of oxidative stress response signaling. These results indicate that the acute toxicity of Maillard reaction products is closely related to their ability to induce oxidative stress, and that DHP-3 is a particularly strong inducer of oxidative stress and thus exhibits high cytotoxicity among Maillard reaction products. In addition, we have shown that a comprehensive analysis comparing multiple Maillard reaction products is effective for elucidating their complex and diverse toxicities.</p>","PeriodicalId":17654,"journal":{"name":"Journal of Toxicological Sciences","volume":"48 5","pages":"263-272"},"PeriodicalIF":2.0,"publicationDate":"2023-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"9396785","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Ephedrine alleviates bleomycin-induced pulmonary fibrosis by inhibiting epithelial-mesenchymal transition and restraining NF-κB signaling.","authors":"Hui Tian,&nbsp;Limei Wang,&nbsp;Taoli Fu","doi":"10.2131/jts.48.547","DOIUrl":"10.2131/jts.48.547","url":null,"abstract":"Pulmonary fibrosis is a lethal and progressive pulmonary disorder in human beings. Ephedrine is a compound isolated from Ephedra and plays a regulatory role in inflammatory response. This study focused on the anti-pulmonary fibrosis effect of ephedrine and its potential molecular mechanism. After a mouse model of pulmonary fibrosis was established through bleomycin (BLM) induction, the survival percentage, body weight, and pulmonary index were measured. Hematoxylin-eosin staining and Masson's trichrome staining for lung tissues were performed to observe the pathological alterations. The viability of lung epithelial BEAS-2B cells, intracellular production of reactive oxygen species, and the levels of pro-inflammatory cytokines were examined by cell counting kit-8 assays, 2',7'-dichlorofluorescein diacetate (DCF-DA) staining, and enzyme-linked immunosorbent assay, respectively. Immunofluorescence staining was performed to determine E-cadherin and vimentin expression after BLM or ephedrine treatment. The mRNA and protein levels of cytokeratin-8, E-cadherin, α-SMA, and vimentin were subjected to quantitative polymerase chain reaction and immunoblotting. Experimental results revealed that ephedrine treatment rescued the repressive impact of BLM on BEAS-2B cell viability, and ephedrine inhibited BLM-induced overproduction of reactive oxygen species and inflammatory response in BEAS-2B cells. Additionally, ephedrine suppressed epithelial-mesenchymal transition (EMT) process stimulated by BLM treatment, as demonstrated by the reduced α-SMA and vimentin levels together with the increased cytokeratin-8 and E-cadherin levels in BLM + Ephedrine group. In addition, ephedrine inhibited NF-κB and activated Nrf-2 signaling in BLM-treated BEAS-2B cells. Moreover, ephedrine ameliorated pulmonary fibrosis in BLM-induced mice and improved the survival of model mice. In conclusion, ephedrine attenuates BLM-evoked pulmonary fibrosis by repressing EMT process via blocking NF-κB signaling and activating Nrf-2 signaling, suggesting that ephedrine might become a potential anti-pulmonary fibrosis agent in the future.","PeriodicalId":17654,"journal":{"name":"Journal of Toxicological Sciences","volume":"48 10","pages":"547-556"},"PeriodicalIF":2.0,"publicationDate":"2023-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"41124681","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Lidocaine induces neurotoxicity in spinal cord neurons in Goto-Kakizaki rats via AMPK-mediated mitophagy.","authors":"Ling Chen,&nbsp;ChenLu Fan,&nbsp;Xuekang Zhang,&nbsp;Shibiao Chen,&nbsp;Lingling Ye,&nbsp;Xiaolan Zheng","doi":"10.2131/jts.48.585","DOIUrl":"https://doi.org/10.2131/jts.48.585","url":null,"abstract":"<p><strong>Objective: </strong>Lidocaine has been reported to induce neurotoxicity, which is further enhanced by high glucose levels. This study is aimed to explore the underlying mechanisms of lidocaine neurotoxicity in spinal cord neurons of diabetes.</p><p><strong>Methods: </strong>Take thirty specific pathogen-free (SPF) healthy Sprague-Dawley (SD) rats and thirty Goto-Kakizaki (GK) rats, aged 12 weeks, weighing 180-200 g. The spinal cord neurons of rats were isolated and cultured in vitro. Cell Counting Kit-8 was used to detect cell proliferation to determine the appropriate concentration and duration of lidocaine. Mitochondrial function was assessed using ATP content, cellular oxygen consumption rate, mitochondrial membrane potential, ROS production, and mitochondrial ultrastructure. Western blot was applied to detect the expression of autophagy- and mitophagy-related molecules PINK1, p-AMPK, LC-3II/LC3-I ratio and mTORC1. Immunofluorescent staining was used to detect the expression of PINK1 and LC3.</p><p><strong>Results: </strong>Lidocaine decreased cell viability of spinal cord neurons in concentration- and time-dependent manners. And lidocaine treatment aggravated mitochondrial dysfunction in GK rats. Furthermore, mitophagy was activated in diabetes, and lidocaine exposure up-regulated mitophagy. AMPK activator MK8722 aggravated mitochondrial damage, increased the expression of PINK1, p-AMPK, LC-3II/LC3-I ratio, and decreased the expression of mTORC1, while AMPK inhibitor Compound C and autophagy inhibitor Bafilomycin A1 reduced mitochondrial damage and decreased the expression of PINK1, p-AMPK, LC-3II/LC3-I ratio, and increased the expression of mTORC1.</p><p><strong>Conclusions: </strong>Lidocaine induced neurotoxicity of spinal cord neurons in GK rats via AMPK-mediated mitophagy.</p>","PeriodicalId":17654,"journal":{"name":"Journal of Toxicological Sciences","volume":"48 11","pages":"585-595"},"PeriodicalIF":2.0,"publicationDate":"2023-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"71424666","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Simulation-based risk assessment for the leakage of toxic substances in a chemical plant and the effects on the human body: ethanol as a working model.","authors":"Akio Horiguchi,&nbsp;Satoshi Numazawa","doi":"10.2131/jts.48.285","DOIUrl":"https://doi.org/10.2131/jts.48.285","url":null,"abstract":"<p><p>Chemical plants must handle a wide variety of hazardous substances. To ensure safety in such plants, it is necessary to conduct extensive and highly accurate risk assessments. In this study, we aimed at developing a method that enables flexible and accurate risk assessment. We combined two different simulation tools to reproduce the phenomena of toxic gas leakage and diffusion as well as its impact on human health. The atmospheric diffusion after the leakage of toxic gas was simulated by computational fluid dynamics (CFD). Assuming the movement line of the person, toxic gas absorption and subsequent metabolism were calculated by a physiologically based pharmacokinetic (PBPK) model. From this, changes in blood concentration of toxic substances with time were simulated and we evaluated the effects of toxic gases on human body. Ethanol was selected as a toxic gas in this study. Based on the assumed scenario, the diffusion of leaked ethanol gas was calculated by CFD leading to the confirmation that the concentration of ethanol gas varies significantly with wind speed, human position, and elapsed time. The PBPK model showed that the maximum blood concentration of ethanol was 161 µmol/L, which is sufficiently low compared to that of ethanol poisoning (i.e., 10,900 µmol/L). These results suggest that the effects on the human body are relatively low and the evacuation can be performed safely. Compared to conventional methods of risk assessment, our new method allows the risk assessment of multiple scenarios, namely interindividual differences, activity status and the used of protective equipment.</p>","PeriodicalId":17654,"journal":{"name":"Journal of Toxicological Sciences","volume":"48 5","pages":"285-298"},"PeriodicalIF":2.0,"publicationDate":"2023-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"9396787","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"The protective effect of thiolutin on doxorubicin-induced H9c2 cardiomyocyte injury.","authors":"Wenyuan Cai,&nbsp;Tingting Teng,&nbsp;Xiaoyan Wang,&nbsp;Baihong Li,&nbsp;Xin Gu,&nbsp;Yafeng Zhou","doi":"10.2131/jts.48.469","DOIUrl":"https://doi.org/10.2131/jts.48.469","url":null,"abstract":"<p><p>The use of doxorubicin (DOX) may contribute to cardiotoxicity, limiting its clinical application. Thiolutin (THL) has been found to exert protective roles in various biological activities, while its effects on DOX-induced cardiotoxicity are still uncovered. Cell counting kit 8 assay was utilized to detect cell viability and half maximal inhibitory concentration of THL in H9c2 cardiomyocytes. The level of lactate dehydrogenase (LDH), adenosine triphosphate (ATP), interleukin (IL)-18 and IL-1 beta (IL-1β) were measured using the corresponding detection kits, and flow cytometry determined cell apoptosis rate. The reactive oxygen species (ROS) accumulation was evaluated by utilizing immunofluorescence or flow cytometry assay. The protein levels of NLR family Pyrin domain 3 (NLRP3), pro-Caspase1, cleaved-Caspase1, gasdermin D (GSDMD) and cleaved-GSDMD (GSDMD-N) in H9c2 cells were detected by immunoblotting assay. The treatment of THL reduced H9c2 cell viability in a gradient-dependent manner. THL treatment reversed the DOX-induced inhibition of proliferation, decrease of ATP, up-regulation of LDH, IL-18, IL-1β and production of ROS, activation of NLRP3 and inflammasome-mediated pyroptosis in H9c2 cells. Additionally, NLRP3 knockdown abolished the effects of THL in DOX-treated H9c2 cells remarkably. This investigation proved that THL notably ameliorated DOX-induced apoptosis, oxidative stress, and pyroptosis in H9c2 cardiomyocytes. Besides, THL effectively inactivated DOX-induced NLRP3 inflammasome in H9c2 cells. These findings revealed a promising drug to assist DOX in its anti-cancer effects and protect the heart of patients.</p>","PeriodicalId":17654,"journal":{"name":"Journal of Toxicological Sciences","volume":"48 8","pages":"469-479"},"PeriodicalIF":2.0,"publicationDate":"2023-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"9935355","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}