The environmental pollutant trichloroethylene (TCE) can induce T-cell-mediated hypersensitivity syndrome. Clinical evidence closely links Epstein-Barr virus (EBV) reactivation to aggravated disease severity, yet the mechanism underlying this chemical-viral synergy remains unknown. Based on the view that hypersensitivity essentially involves sustained activation of effector T-cells and disruption of immune homeostasis, this study aims to investigate how EBV reactivation cooperates with TCE metabolite trichloroethanol (TCOH) to regulate the T-cell apoptosis process by affecting mitochondrial dynamics and autophagy, thereby exacerbating trichloroethylene-induced hypersensitive syndrome (TIHS). Results show that TCOH exposure induces mitochondrial damage and apoptosis in T-cells. However, co-exposure with EBV significantly alleviates TCOH-induced mitochondrial damage by enhancing Drp1-mediated mitochondrial fission and promoting subsequent PINK1/Parkin pathway-dependent mitophagy, which in turn suppresses T-cell apoptosis. In an animal model, administration of the Drp1 inhibitor Mdivi-1 inhibited mitophagy, increased T-cell apoptosis, and effectively alleviated skin and hepatic immune injury in TCE-sensitized mice. This confirms that Drp1 is a key molecular target regulating the T-cell apoptosis program and influencing the progression of TIHS. The study reveals that during TIHS development, EBV reactivation exploits Drp1 to promote mitochondrial fission and mitophagy, thereby hindering the normal apoptotic clearance of activated effector T-cells, leading to their sustained activation and immune attack, which exacerbates tissue inflammation and injury. This finding provides a new scientific perspective for understanding the mechanism by which viral co-exposure aggravates TIHS.

Cobalt is a critical raw material for consumer electronic products such as electric vehicles and mobile communication devices. Currently, its recovery rate remains relatively low, resulting in a large amount of waste cobalt entering soil and groundwater environments, which poses a potential threat to human health. Previous studies have mostly focused on the effects of acute cobalt exposure. However, the in vivo metabolic mechanism of long-term low-dose cobalt exposure remains unclear, and the long-term toxic effects of cobalt on the heart await further investigation. Based on the existing model framework, this study constructed a cobalt Physiologically based pharmacokinetic (PBPK) model incorporating a heart compartment, filling the research gap in this field. Meanwhile, the study calibrated the parameters of the PBPK model by integrating the specific exposure scenarios and dietary exposure characteristics of the Chinese population, and conducted model evaluation and evaluation using epidemiological data of the Chinese population. With the help of Monte Carlo simulation (MCS) technology, this study conducted an in-depth analysis of the inter-individual exposure and metabolic characteristics. The results showed that the deviation between the cobalt concentrations in blood, urine, liver, kidneys, and heart of the Chinese population predicted by the model and the measured values was basically controlled within a 2-fold range. The cobalt PBPK model constructed in this study provides a key tool for the accurate risk assessment and practical application of cobalt exposure in the Chinese population, and also offers a referable research paradigm for accurate risk assessment under the specific characteristics of different ethnic groups.

Abstract Long-term lithium exposure may lead to chronic kidney disease (CKD), but its mode of action (MOA) and biomarkers remain unclear. This study aimed to explore the potential mechanisms by integrating an adverse outcome pathway (AOP) framework and machine learning. A chemical-gene-phenotype-disease network (CGPDN) was used to construct an AOP framework through network toxicology. Drug enrichment analysis, molecular docking, and in vitro experiments were conducted to identify potential targeted drugs. Moreover, machine learning was used to identify potential noninvasive biomarkers. The AOP network was constructed from 14 key targets and 12 key phenotypes that were screened via CGPDN. In this AOP framework, “response to metal ion” was characterized as the molecular initiating event. Furthermore, “oxidative stress” and “Increase, Cell death” were identified as central key events. Drug enrichment analysis and molecular docking revealed that simvastatin is a potential target drug. In addition, lithium exposure induced abnormal expression of 7 key targets in HK-2 cells, while simvastatin pretreatment reversed the changes in these targets and reduced renal toxicity. The expression patterns of FAS, HSP90B1, and CASP1 were consistent with those observed in the CKD kidney and PBMC transcriptomic data sets. Further, a machine learning model and Shapley Additive exPlanations analysis confirmed that the three targets were potential blood biomarkers of lithium-induced CKD. This study developed a novel paradigm integrating AOP with machine learning, which elucidated the MOA of lithium-induced CKD and filled the gap in early-warning biomarkers. Our findings reveal simvastatin as a promising intervention drug and provide a feasible framework for early risk assessment in lithium-exposed populations.

Little is known about how diesel engine exhaust (DEE) exposure is associated with changes in blood lipid concentrations, as well as the molecular initiating events (MIEs) and mechanisms underlying this association. The aim of this research was to determine the MIEs and key molecular events and thus construct a putative adverse outcome pathway (AOP) from DEE exposure to an increased risk of metabolism-related diseases by integrating occupational epidemiology and metabolomics analyses. We recruited 136 participants with high DEE exposure and 99 healthy controls to investigate the associations between polycyclic aromatic hydrocarbons (PAHs; an exposure biomarker of DEE), blood lipids (high-density lipoprotein cholesterol (HDL-C) and lowdensity lipoprotein cholesterol), and plasma metabolites. Our results revealed that the concentrations of six OH-PAHs were increased significantly in urine samples from participants with high DEE exposure (P <= 0.001), and that 9-hydroxyphenanthrene (9-OHPh) is uniquely associated with decreased HDL-C concentrations (P = 0.034). Each 10% increase in the 9-OHPh concentration corresponded to a 0.225% decrease in the HDL-C concentration. Targeted metabolomics analysis revealed key pathways associated with both 9-OHPh and HDL-C, including alanine and glutamine metabolism, glutathione biosynthesis, sirtuin, and TP53 pathway. Additionally, nine metabolites (e.g. glutamate, glutamine, and glutathione) were enriched in pathways associated with both 9OHPh and HDL-C. Bioinformatic analyses of these metabolites identified potential MIEs (e.g. CPT1B, LDHB, and SIX1) and adverse outcomes (liver damage, renal necrosis, and cardiac damage). Based on these findings, we propose for the first time a putative AOP connecting DEE exposure with metabolism-related disease risks, which is driven by alterations in MIEs, subsequent metabolic disturbances, and reduced HDL-C levels, thus providing crucial insights into environmental contributors to metabolic disorders.

BACKGROUND:Systemic immune inflammation index (SII) and systemic inflammation response index (SIRI) have recently emerged as more comprehensive indicators of systemic inflammation. However, few studies have been conducted to evaluate the effect of serum cotinine levels and weight-adjusted waist index (WWI) with SII and SIRI. This study aims to evaluate the independent and combined impact of serum cotinine levels and WWI with SII and SIRI. METHODS:This study enrolled 9,099 adults based on the National Health and Nutrition Examination Survey 2013-2016. Generalized linear models were employed to assess the associations between cotinine exposure and WWI with SII and SIRI. Moreover, smoothing curve fitting was conducted to investigate the potential nonlinear relationships between the WWI with SII and SIRI levels. Additionally, subgroup analyses and interaction tests were conducted. The combined associations of the WWI and cotinine with systemic inflammatory indices were further assessed. RESULTS:High serum cotinine exposure was associated with increased SII and SIRI. The WWI was positively correlated with SII and SIRI. Smoothing curve fitting demonstrated a nonlinearly increasing dose-response connection between WWI and SII and SIRI. Stratified analyses also showed a positive association of cotinine levels and WWI with the systemic inflammatory indices across subgroups. Furthermore, a positive joint association was observed between serum cotinine level and WWI with SII and SIRI. CONCLUSIONS:These findings suggest that serum cotinine levels and WWI may be positively related to SII and SIRI. Serum cotinine levels and WWI may have a positive combined effect.

Lithium nickel manganese cobalt oxide (NMC) is a widely used cathode material in lithium-ion batteries. However, its increasing utilization has raised concerns about environmental contamination and potential health risks, underscoring the need for comprehensive toxicological assessments. In this study, we evaluated the tissue distribution and toxicological effects of NMC in adult male C57BL/6J mice following 28-day intratracheal instillation at doses of 32.5, 162.5, and 325 μg/kg/day. Our findings demonstrated that lithium primarily accumulated in the heart, while nickel also showed high accumulation ability in the heart, followed by the liver and epididymis. Cobalt accumulated predominantly in the spleen, kidneys, and epididymis, whereas manganese was mainly concentrated in the epididymis. Notably, exposure to high doses of NMC resulted in a reduction in body weight, elevated levels of systemic inflammation biomarkers, and significant histopathological changes. These histopathological alterations included alveolar fusion and structural damage in the lungs, loss of distinction between red and white pulp in the spleen, renal tubular dilation, and a decrease in spermatogenic cells in the testes and epididymis. Furthermore, NMC exposure disrupted the homeostasis of trace metals (copper, vanadium, molybdenum, strontium, selenium, and stannum) in mice, suggesting their potential roles in NMC-induced toxicity. This study provides a detailed characterization of the distribution and toxic effects of low-dose NMC exposure, contributing valuable data for the toxicology evaluation of NMC materials.

Trichloroethylene (TCE), extensively used as an organic solvent in various industrial applications, has been identified as a causative factor in inducing hypersensitivity syndrome (THS). Currently, there is no specific treatment for THS, and most patients experience serious adverse outcomes due to extensive skin damage leading to severe infection. However, the pathogenesis of THS-associated skin damage remains unclear. This study aims to elucidate the mechanism underlying skin damage from the perspective of intercellular communication and gap junctions in THS. Our results verified that hyperactivation of connexin43 gap junctions, caused by the aberrantly elevated expression of connexin43, triggers a bystander effect that promotes apoptosis and inflammation in THS via the TNF-TNFRSF1B and mitochondria-associated pathways. Additionally, we identified the gap junction inhibitor Carbenoxolone disodium (CBX) as a promising agent for the treatment of skin damage in THS. CBX protects against inflammatory cell infiltration in the skin and decreases immune cell imbalance in the peripheral blood of THS mice. Furthermore, CBX reduces connexin43 expression, apoptosis and inflammation in THS mice. The study reveals new insights into the mechanisms underlying TCE-induced skin damage, offering a potential treatment strategy for the development of effective therapies targeting severe dermatitis induced by chemical exposure.

BACKGROUND:The weight-adjusted waist index (WWI) is a recently developed obesity metric, and the aim of this study was to investigate the relationship between physical activity (PA) and WWI and the homeostasis model assessment of insulin resistance (HOMA-IR) in adolescents, as well as the joint association of HOMA-IR. METHODS:This study was based on the National Health and Nutrition Survey conducted between 2013 and 2016 and included 1024 adolescents whose median age was 15.4. Multivariate linear regression was used to examine the associations between HOMA-IR and PA and WWI. Using generalized additive models, a potential nonlinear link between WWI and HOMA-IR was evaluated. Subgroup analysis was also carried out. RESULTS:The fully adjusted model revealed a positive association (β: 0.48, 95% CI: 0.43, 0.53) between the WWI and HOMA-IR. The HOMA-IR was lower in physically active (β: -0.16, 95% CI: -0.26, -0.05) participants versus inactive participants. Participants who had higher WWI and were not physically active (β: 0.69; 95% CI: 0.56, 0.82) had the highest levels of HOMA-IR compared to participants who had lower WWI and were physically active. Subgroup analysis revealed that these correlations were similar in males and females. CONCLUSION:Our results demonstrated that higher WWI and PA were associated with a lower HOMA-IR and that WWI and PA had a combined association with HOMA-IR. The findings of this study are informative for the preventing insulin resistance in adolescents.

Systematic studies on the associations between co-exposure to multiple metals and chronic kidney disease (CKD), as well as the underlying mechanisms, remain insufficient. This study aimed to provide a comprehensive perspective on the risk of CKD induced by multiple metal co-exposures through the integration of occupational epidemiology and adverse outcome pathway (AOP). The study participants included 401 male mine workers whose blood metal, β2-microglobulin (β2-MG), and cystatin C (Cys-C) levels were measured. Generalized linear models (GLMs), quantile g-computation models (qgcomp), least absolute shrinkage and selection operator (LASSO), and bayesian kernel machine regression (BKMR) were utilized to identify critical nephrotoxic metals. The mean concentrations of lead, cadmium, mercury, arsenic, and manganese were 191.93, 3.92, 4.66, 3.11, 11.35, and 16.33 µg/L, respectively. GLM, LASSO, qgcomp, and BKMR models consistently identified lead, cadmium, mercury, and arsenic as the primary contributors to kidney toxicity. Based on our epidemiological analysis, we used a computational toxicology method to construct a chemical-genetic-phenotype-disease network (CGPDN) from the Comparative Toxicogenomics Database (CTD), DisGeNET, and GeneCard databases, and further linked key events (KEs) related to kidney toxicity from the AOP-Wiki and PubMed databases. Finally, an AOP framework of multiple metals was constructed by integrating the common molecular initiating events (reactive oxygen species) and KEs (MAPK signaling pathway, oxidative stress, mitochondrial dysfunction, DNA damage, inflammation, hypertension, cell death, and kidney toxicity). This is the first AOP network to elucidate the internal association between multiple metal co-exposures and CKD, providing a crucial basis for the risk assessment of multiple metal co-exposures.

Trichloroethylene (TCE)-induced hypersensitivity syndrome (THS) has been a concern for many researchers in the field of environmental and occupational health. Currently, there is no specific treatment for THS, leaving patients to contend with severe infections arising from extensive skin lesions, consequently leading to serious adverse effects. However, the pathogenesis of severe skin damage in THS remains unclear. This study aims to investigate the specific danger signals and mechanisms underlying skin damage in THS through in vivo and in vitro experiments. We identified that cell supernatant containing 15 kDa granulysin (GNLY), released from activated CD3-CD56+NK cells or CD3+CD56+NKT cells in PBMC induced by TCE or its metabolite, promoted apoptosis in HaCaT cells. The apoptosis level decreased upon neutralization of GNLY in the supernatant by a GNLY-neutralizing antibody in HaCaT cells. Subcutaneous injection of recombinant 15 kDa GNLY exacerbated skin damage in the THS mouse model and better mimicked patients’ disease states. Recombinant 15 kDa GNLY could directly induce cellular communication disorders, inflammation, and apoptosis in HaCaT cells. In addition to its cytotoxic effects, GNLY released from TCE-activated NK cells and NKT cells or synthesized GNLY alone could induce aberrant expression of the E3 ubiquitin ligase PDZRN3, causing dysregulation of the ubiquitination of the cell itself. Consequently, this resulted in the persistent opening of gap junctions composed of connexin43, thereby intensifying cellular inflammation and apoptosis through the “bystander effect”. This study provides experimental evidence elucidating the mechanisms of THS skin damage and offers a novel theoretical foundation for the development of effective therapies targeting severe dermatitis induced by chemicals or drugs.

Due to the complexity of environmental exposure factors and the low levels of exposure in the general population, identifying the key environmental factors associated with diabetes and understanding their potential mechanisms present significant challenges. This study aimed to identify key polycyclic aromatic hydrocarbons (PAHs) contributing to increased fasting blood glucose (FBG) concentrations and to explore their potential metabolic mechanisms. We recruited a highly PAH-exposed diesel engine exhaust testing population and healthy controls. Our findings found a positive association between FBG concentrations and PAH metabolites, identifying 1-OHNa, 2-OHPh, and 9-OHPh as major contributors to the rise in FBG concentrations induced by PAH mixtures. Specifically, each 10 % increase in 1-OHNa, 2-OHPh, and 9-OHPh concentrations led to increases in FBG concentrations of 0.201 %, 0.261 %, and 0.268 %, respectively. Targeted metabolomics analysis revealed significant alterations in metabolic pathways among those exposed to high levels of PAHs, including sirtuin signaling, asparagine metabolism, and proline metabolism pathway. Toxic function analysis highlighted differential metabolites involved in various dysglycemia-related conditions, such as cardiac arrhythmia and renal damage. Mediation analysis revealed that 2-aminooctanoic acid mediated the FBG elevation induced by 2-OHPh, while 2-hydroxyphenylacetic acid and hypoxanthine acted as partial suppressors. Notably, 2-aminooctanoic acid was identified as a crucial intermediary metabolic biomarker, mediating significant portions of the associations between the multiple different structures of OH-PAHs and elevated FBG concentrations, accounting for 16.73 %, 10.84 %, 10.00 %, and 11.90 % of these effects for 1-OHPyr, 2-OHFlu, the sum concentrations of 2- and 9-OHPh, and the sum concentrations of total OH-PAHs, respectively. Overall, our study explored the potential metabolic mechanisms underlying the elevated FBG induced by PAHs and identified 2-aminooctanoic acid as a pivotal metabolic biomarker, presenting a potential target for intervention.

Trichloroethylene-induced hypersensitivity dermatitis (TIHD) is a delayed hypersensitivity response that is affected by genetic and environmental factors. Occupational exposure to trichloroethylene (TCE) enhances antigen presentation, leading to hypersensitivity in workers with the HLA-B*13:01 allele. Several studies have observed the activation of herpesviruses, such as EpsteinBarr virus (EBV), in TIHD patients. However, the underlying mechanisms remain unclear. Toll-like receptors (TLRs) play a pivotal role in the pathogenesis of herpesvirus infection. This study aimed to explore whether TLRs serve as a shared mechanism for both herpesvirus and allergenic chemicals. In this study, HLA-B*13:01-transfected Hmy2. A C1R cell model was constructed, and cells were treated with TCOH and EBV to explore the possible mechanisms. We established a mouse model of dermatitis and used a TLR4 agonist to verify the effect of herpesvirus on TIHD. The results showed that EBV and TCOH synergistically enhance antigen processing and presentation via the TLR2/NF-κB axis. Furthermore, TLR4 agonist further aggravated skin lesions and liver damage in TCE-sensitized mice through TLR4/NF-κB axis-mediated antigen processing and presentation. Together, this study indicates that viral infection further aggravates the inflammatory response in TIHD based on environment-gene interactions.

BACKGROUND:The association between single metal exposure and chronic kidney disease (CKD) has been established. However, there is limited research on the effects of multi-metal mixtures and their potential age-specific associations with kidney injury. This study aimed to examine the relationship between metal mixtures and kidney function in adults, while also exploring the modifying effects of age. METHODS:We included a subset (n = 4250) of a nationally representative adult population in the National Health and Nutrition Examination Survey (NHANES) 2013-2016. Kidney function was assessed using the glomerular filtration rate (eGFR) and creatinine albumin ratio (ACR). The individual and combined effects of lead (Pb), cadmium (Cd), mercury, and manganese on kidney injury and the risk of CKD were evaluated. RESULTS:Pb and Cd were found to be positively associated with decreased kidney function. For a one Ln-unit increase in lead and cadmium, the adjusted ORs of CKD were 1.60 (95% CI: 1.35, 1.90) and 1.41 (95% CI:1.12, 1.77), respectively. We also observed an interaction between lead and cadmium for ACR. We also observed the joint effect between Pb and Cd on eGFR, ACR and CKD. Stratified analysis found a higher risk of decreased kidney function among older individuals. The quantile-g calculation model further showed that metal mixture was associated with decreased kidney function and the risk of CKD (OR = 1.53, 95% CI: 1.22, 1.90). And lead and cadmium were the main contributors. And Pb and Cd were the major components that increased the risk of CKD. CONCLUSION:Co-exposure to metal mixture were associated with reduced kidney function in adults, especially in older. Our findings support co-exposure to lead and cadmium as risk factors of CKD in adults.

Abstract Background Glyphosate is a commonly used herbicide worldwide and is purportedly associated with multiple health effects. Research assessing the association of glyphosate concentrations with glycosylated hemoglobin (HbA1c) levels and the prevalence of diabetes is scarce. We sought to evaluate the association between urinary glyphosate levels and HbA1c levels and the prevalence of diabetes. Methods A total of 2,745 adults in the National Health and Nutrition Examination Survey from 2013 to 2016 were included in this study. Generalized linear models (GLM) were applied to evaluate the associations of glyphosate concentrations with HbA1c levels and the prevalence of diabetes. The dose–response relationship was examined using restricted cubic splines (RCS). Results Significantly positive correlations of urinary glyphosate concentrations with HbA1c levels (percentage change: 1.45; 95% CI: 0.95, 1.96; P < 0.001) and the prevalence of diabetes (OR: 1.45; 95% CI: 1.24, 1.68; P < 0.001) were found after adjustment. Compared with the lowest quartile of glyphosate levels, the highest quartile was positively associated with HbA1c levels (percentage change: 4.19; 95% CI: 2.54, 5.85; P < 0.001) and the prevalence of diabetes (OR: 1.89; 95% CI: 1.37, 2.63; P < 0.001). The RCS curves demonstrated a monotonically increasing dose–response relationship between urinary glyphosate levels and the prevalence of diabetes and HbA1c levels. Conclusions Urinary glyphosate concentrations are positively associated with HBA1c levels and the prevalence of diabetes. To verify our findings, additional large-scale prospective investigations are required.

Trichloroethylene hypersensitivity syndrome (THS), mainly caused by occupational exposure to trichloroethylene (TCE), can give rise to serious and fatal hepatic damage. To date, the precise mechanisms of hepatic damage in THS remain unclear. Recent studies showed that reactive oxygen species (ROS) play a core role in cell death and inflammatory response. Therefore, the present study sought to explore whether ROS-mediated inflammatory responses contribute to the hepatic damage in TCE sensitization. To this end, a mouse model of TCE sensitization was established; in some cases, hosts were pretreated with tempol, an ROS scavenger. The results showed that TCE sensitization caused hepatic pathological/functional changes, ROS generation, and oxidative stress, alterations of the anti-oxidant defense Nrf2/HO-1/NLRP3 pathway, and pro-inflammatory cytokine formation in the liver. ROS scavenging via pretreatment with tempol was found not only to inhibit the hepatic oxidative stress, but also to regulate Nrf2/HO-1/NLRP3 pathway activity. In all cases, tempol was able to mitigate the pathologic changes induced by TCE sensitization. In summary, the results here demonstrated a novel molecular mechanism wherein ROS-mediated inflammatory responses play a central role in TCE-induced liver damage. Therapies targeting ROS scavenging could help to protect against hepatic damage by regulating Nrf2/HO-1/NLRP3 pathway activities in TCE-sensitized hosts.

Trichloroethylene (TCE), a commonly used organic solvent, is known to cause trichloroethylene hypersensitivity syndrome (THS), also called occupational medicamentosa–like dermatitis due to TCE (OMDT) in China. OMDT patients presented with severe inflammatory kidney damage, and we have previously shown that the renal damage is related to the terminal complement complex C5b-9. Here, we sought to determine whether C5b-9 participated in TCE-induced immune kidney injury by promoting pyroptosis, a new form of programed cell death linked to inflammatory response, with underlying molecular mechanisms involving the NLRP3 inflammasome. A BALB/c mouse-based model of OMDT was established by dermal TCE sensitization in the presence or absence of C5b-9 inhibitor (sCD59-Cys, 25μg/mouse) and NLRP3 antagonist (MCC950, 10 mg/kg). Kidney histopathology, renal function, expression of inflammatory mediators and the pyroptosis executive protein gasdermin D (GSDMD), and the activation of pyroptosis canonical NLRP3/caspase-1 pathway were examined in the mouse model. Renal tubular damage was observed in TCE-sensitized mice. GSDMD was mainly expressed on renal tubular epithelial cells (RTECs). The caspase-1–dependent canonical pathway of pyroptosis was activated in TCE-induced renal damage. Pharmacological inhibition of C5b-9 could restrain the caspase-1–dependent canonical pathway and rescued the renal tubular damage. Taken together, our results demonstrated that complement C5b-9 plays a central role in TCE-induced immune kidney damage, and the underlying mechanisms involve NLRP3-mediated pyroptosis.

[背景]三氯乙烯(TCE)可通过受污染的水或空气经生物蓄积作用进入人体,从而产生健康危害,受累脏器以肝脏最为多见.[目的]观察TCE经口暴露小鼠肝脏中M1型库普弗细胞(KCs)极化的情况,探讨组蛋白赖氨酸去甲基化酶JMJD3与KCs M1型极化之间的关系.[方法]选取72只SPF级雌性BALB/c小鼠,6～8周龄,将其随机分为4组:空白对照组(18只)、溶剂对照组(18只)、2.5 mg·mL–1 TCE组(18只)和5.0 mg·mL–1 TCE组(18只),适应性喂养1周.按照课题组前期研究建立小鼠TCE经口暴露模型,染毒8周.分别在第2、4、8周时处死小鼠取肝脏组织,采用Western blotting法检测小鼠肝脏组织中组蛋白赖氨酸去甲基化酶JMJD3的蛋白表达水平,采用免疫荧光法对巨噬细胞标志F4/80和M1型巨噬细胞表面标志CD11c进行共定位,采用免疫组织化学法检测巨噬细胞M1型标志CD16/32和小鼠肝脏M1型巨噬细胞相关炎性因子肿瘤坏死因子-α(TNF-α)的表达情况.[结果]在第2、4、8周时,各组小鼠状态良好,没有出现因TCE染毒而死亡的个体,各组间小鼠耗水量差异无统计学意义,各时间点小鼠体重增长及肝脏系数的组间差异没有统计学意义(P>0.05).Western blotting法检测结果显示:在各时间点空白对照组和溶剂对照组JMJD3蛋白表达水平差异均无统计学意义,2.5 mg·mL–1 TCE组和5.0 mg·mL–1 TCE组JMJD3蛋白表达水平相较于对照组均升高,且第4、8周5.0 mg·mL–1 TCE组JMJD3蛋白表达水平高于2.5 mg·mL–1 TCE组(P<0.05).免疫荧光共定位结果显示:在各时间点空白对照组和溶剂对照组F4/80与CD11c表达较少,2.5 mg·mL–1 TCE组和5.0 mg·mL–1 TCE组F4/80与CD11c表达较高.免疫组织化学检测结果显示:CD16/32和TNF-α在空白对照组和溶剂对照组表达较低,而在2.5 mg·mL–1 TCE组和5.0 mg·mL?1 TCE组出现大量沉积.[结论]KCs M1型极化及促炎因子的表达可能与经口暴露TCE所致的JMJD3表达水平增加有关.

Objective: To explore the mechanism of reactive oxygen species/thioredoxin-interacting protein/nucleotide-binding oligomerization domain-like receptor 3 (ROS/TXNIP/NLRP3) pathway in the skin injury of trichloroethylene (TCE) sensitized mice. Methods: In August 2020, 40 female BALB/c mice were randomly divided into control group (n=5) , solvent control group (n=5) , TCE treatment group (n=15) and TCE+(2-(2, 2, 6, 6-Tetrameyhylpiperidin-1-oxyl-4-ylamino)-2-oxoethyl) triphenylphosphonium chloride (Mito TEMPO) treatment group (n=15) . The TCE sensitization model was established. Mice in the TCE treatment group and TCE+Mito TEMPO treatment group were divided into the sensitized positive group and the sensitized negative group according to the skin erythema and edema reactions on the back of the mice 24 h after the last stimulation. The mice were sacrificed 72 h after the last stimulation, the back skin of the mice was taken, and the skin lesions were observed. Immunohistochemistry (IHC) was used to detect the expression level of NLRP3, and the Western Blot was performed to detect the expression levels of NLRP3, apoptosis-associated speck-like protein containing a CARD (ASC) , cysteinyl aspartate specific proteinase 1 (Caspase 1) , Interleukin-1β (IL-1β) and TXNIP proteins in the skin of the mice, the reactive oxygen species (ROS) kit was used to detect the level of intracellular ROS in the back skin tissue. Results: The sensitization rates of TCE treatment group and TCE+Mito TEMPO treatment group were 40.0% (6/15) and 33.3% (5/15) , respectively, and there was no significant difference between the two groups (P>0.05) . The back skin of the mice in the TCE sensitized positive group was thickened and infiltrated by a large number of inflammatory cells. The number of mitochondria in the epidermis cells was significantly reduced, the mitochondrial crest disappeared and vacuolar degeneration occurred. TCE+Mito TEMPO sensitized positive group had less damage, more mitochondria and relatively normal cell structure. Compared with the solvent control group and corresponding sensitized negative groups, the expression levels of NLRP3, ASC, Caspase 1, IL-1β, TXNIP proteins and the content of ROS in the TCE sensitized positive group and TCE+Mito TEMPO sensitized positive group were significantly increased (P<0.05) . Compared with TCE sensitized positive group, the expression levels of NLRP3, ASC, Caspase 1, IL-1β, TXNIP proteins and the content of ROS in the TCE+Mito TEMPO sensitized positive group were significantly decreased (P<0.05) . Conclusion: ROS/TXNIP/NLRP3 pathway was activated and then encouraged the release of IL-1β, finally aggravated the TCE-induced skin injury.

目的 探讨组织蛋白酶L(CTSL)介导三氯乙烯(TCE)致敏小鼠肾脏损伤的分子机制.方法 41只雌性BALB/c小鼠随机分为空白组(5只)和溶剂组(5只),TCE处理组(15只)和TCE+CTSLi处理组(16只),建立TCE经皮致敏小鼠模型,对TCE+CTSLi处理组小鼠腹腔注射CTSL抑制剂(10 mg/kg)进行预处理,根据小鼠皮肤致敏评分评估为阳性组和阴性组.通过电镜和HE染色观察小鼠肾脏病理情况,血清尿素氮(BUN)评估小鼠肾功能水平,免疫荧光检测CTSL的表达情况,TUNEL染色评估小鼠肾脏细胞凋亡情况,免疫印迹法检测肾脏蛋白激酶C(PKC)信号分子的活化情况.结果 TCE处理组和TCE+CTSLi处理组的致敏率分别为53.3％(8/15)和50.0％(8/16),致敏率差异无统计学差异(P>0.05).病理学结果表明TCE致敏阳性小鼠出现肾小管细胞水肿、空泡变性,肾小球基底膜增厚,足细胞足突融合,线粒体空泡样变性等表现.肾功能结果表明TCE致敏阳性小鼠血清BUN水平较其他组升高.免疫荧光结果表明CTSL在TCE致敏阳性小鼠肾脏表达水平升高(P<0.05,F=82.438),肾脏结构细胞凋亡水平同样高于其他各组(P<0.05).免疫印迹结果表明TCE致敏阳性小鼠肾脏中PKC蛋白磷酸化水平升高,而使用CTSLi预处理后,TCE+CTSLi致敏阳性组中的表达出现下调(P<0.05,F=35.686),肾脏细胞凋亡水平降低,肾脏损害得以改善.结论 CTSL可能通过激活PKC信号介导肾脏细胞凋亡,加重TCE致敏小鼠肾脏损害.

Trichloroethylene (TCE) hypersensitivity syndrome (THS), called occupational medicamentosa-like dermatitis due to TCE (OMDT) in China, is a fatal occupational disorder caused by TCE exposure. Visceral damage, including kidney injury, is one of the major complications. Necroptosis is a regulated cell death form linked to local inflammatory response. This study aimed to investigate whether renal cell necroptosis was involved in TCE-induced kidney injury. A Balb/c mouse model of TCE sensitization was utilized to study mechanisms of modulation of TCE-induced renal necroptosis. Renal histology (using light and transmission electron microscopy) and renal tubular impairment indexes, including α1-microglobulin (α1-MG), and β2-microglobulin (β2-MG), were evaluated. In addition, tissue expression of necroptosis-related proteins, including tumor necrosis factor (TNF)-α, TNF receptor 1 (TNFR1), receptor-interacting protein kinase 3 (RIPK3), p-RIK3, mixed lineage kinase domain-like protein (MLKL), and p-MLKL, were also evaluated. The study here confirmed TCE sensitization caused damage to renal tubules and renal tubular epithelial cell (RTEC) necroptosis. In mice treated with R7050 (a specific TNFα antagonist), it was also seen that inhibition of TNFα expression could effectively inhibit RTEC necroptosis and improve renal function in the TCE-sensitized mice. Taken together, these results help to define a novel mechanism by which RTEC necroptosis plays a key role in TCE-induced kidney damage.