Deoxynivalenol (DON), a toxic mycotoxin produced by Fusarium Fungi, poses substantial risks to both human and animal health. Although its male reproductive toxicity has been reported, the underlying molecular mechanisms and potential therapeutic strategies remain insufficiently elucidated. In this study, we investigated the effects of DON on testicular injury in mice and explored the associated molecular pathways. DON exposure markedly reduced testicular sperm count, impaired spermatogenesis, and caused structural damage to the seminiferous tubules. Furthermore, DON elicited pronounced inflammatory responses in both testicular tissues and TM4 mouse Sertoli cells. Mechanistically, DON induced mitochondrial damage, resulting in mitochondrial DNA (mtDNA) leakage into the cytoplasm. The cytosolic mtDNA subsequently activated the cyclic GMP-AMP (cGAS)stimulator of interferon genes (STING) signaling pathway, promoting TANK-binding kinase 1 (TBK1) and interferon regulatory factor 3 (IRF3) phosphorylation and enhancing the expression of pro-inflammatory cytokines, including IL-6 and TNF-alpha, via NF-kappa B activation. Notably, depletion of mtDNA with ethidium bromide (EtBr) or pharmacological inhibition of STING with H151 attenuated DON-induced inflammation, whereas mtDNA transfection further amplified cGAS-STING activation. Collectively, these findings demonstrate that mtDNA leakage-mediated activation of the cGAS-STING pathway plays a central role in DON-induced testicular inflammation and identify potential therapeutic targets for mitigating DON-associated male reproductive toxicity.

T-2 toxin, a type A trichothecene mycotoxin, induces severe cytotoxicity primarily through mitochondrial dysfunction and apoptosis. In this study, the protective role of astragaloside IV (AS-IV) against T-2 toxin-induced damage was investigated using the human hepatoma cell line HepG2 as an in vitro model. Our results demonstrated that T-2 toxin significantly reduced cell viability, promoted reactive oxygen species (ROS) accumulation, and impaired mitochondrial function, as evidenced by disrupted respiratory activity, loss of mitochondrial membrane potential, and reduced ATP production. These alterations were accompanied by dysregulated mitochondrial homeostasis, characterized by enhanced mitochondrial biogenesis and excessive mitochondrial fission, along with activation of the mitochondrial apoptotic pathway. Pretreatment with AS-IV mitigated ROS accumulation, rescued impaired mitochondrial respiratory function, stabilized mitochondrial membrane potential, maintained cellular ATP level, and restored mitochondrial homeostasis. Furthermore, AS-IV suppressed T-2 toxin-triggered apoptosis by modulating the BAX/BCL-2 ratio, inhibiting cytochrome c release, and attenuating caspase activation. Together, these findings indicate that AS-IV protects HepG2 cells from T-2 toxin-induced cytotoxicity by maintaining mitochondrial homeostasis and suppressing the intrinsic apoptotic pathway, providing mechanistic evidence for its protective effect.

Deoxynivalenol (DON) pollution remains a significant safety concern for humans and animals due to its widespread occurrence in grains and feedstuffs. Copper (Cu) plays a vital role as a trace element in maintaining biological functions in humans and animals. Nevertheless, the risks of DON and Cu co-exposure and the underlying mechanisms of intestinal toxicity remain unclear. Herein, IPEC-J2 cells and a mouse intestinal organoid model were used to evaluate the combined toxic effects of co-exposure to DON and Cu. Co-exposure to DON and Cu at low doses has significant synergistic toxic effects, which disrupt the intestinal barrier and significantly inhibit intestinal organoid development. Mechanistically, co-treatment with DON and Cu alters the levels of apoptosis and ferroptosis related proteins, and consequently activates apoptosis and ferroptosis in mouse intestinal organoids and IPEC-J2 cells. These activations are driven by DON + Cu exposure-induced mtROS accumulation, disrupted MMP and mPTP, and inhibited the expression of mitochondrial antioxidant proteins. Pretreatment with the mtROS-targeted scavenger MitoQ effectively assuaged DON + Cu exposure-caused mitochondrial injury and subsequently attenuated the apoptosis and ferroptosis. Overall, our research revealed the synergistic toxicity of DON + Cu exposure and confirmed the potential mechanism of DON + Cu exposure-induced intestinal toxicity, which will provide valuable insights into their potential combined toxic effects on humans and animals.

With the development of Type 1 diabetes mellitus (T1DM), various complications can be caused. Hyperglycemia affects the microenvironment of cardiomyocytes, changes endoplasmic reticulum homeostasis, triggers unfolding protein response and eventually promotes myocardial apoptosis. However, insulin therapy alone cannot effectively combat the complications caused by T1DM. Forty adult beagles were randomly divided into five groups: control group, diabetes mellitus group, insulin group, insulin combined with NAC group, and NAC group. 24-hour blood glucose, 120-day blood glucose, 120-day body weight, and serum FMN content were observed, furthermore, hematoxylin-eosin staining, Periodic acid Schiff reagent staining, and Sirius red staining of the myocardium were evaluated. The protein expressions of GRP78, ATF6, IRE1, PERK, JNK, CHOP, caspase 3, Bcl2, and Bax were detected. Results of the pathological section of myocardial tissue indicated that insulin combined with NAC therapy could improve myocardial pathological injury and glycogen deposition. Additionally, insulin combined with NAC therapy down-regulates the expression of GRP78, ATF6, IRE1, PERK, JNK, CHOP, caspase3, and Bax. These findings suggest that NAC has a phylactic effect on myocardial injury in beagles with T1DM, and the mechanism may be related to the improvement of endoplasmic reticulum stress-induced apoptosis.

To study the effects of Cu overload on ER quality control in duck cerebrums, 144 ducks were treated with 8 mg/kg, 100 mg/kg, 200 mg/kg and 400 mg/kg Cu added in the feed for 45 days. From histopathological examination, we found that excessive Cu increased the amount of microglia and disintegrated neuron, decreased the number of Nissl bodies, perturbed nerve fibers in duck cerebrums. Cu poisoning also increased Cu, H2O2, T-SOD, and MDA levels, decreased Fe and CAT contents in duck cerebrums. Furthermore, Cu treatment upregulated the mRNA levels of the unfolded protein response genes (PERK, ATF6, and IRE1), ER-associated degradation genes (CNX, Derlin1, and Derlin2), autophagy genes (ATG5, ATG7, ATG10, Beclin1, LC3A, LC3B, and P62), and heat shock response genes (Hsp70 and Hsp90) in duck cerebrums; elevated the protein levels of p-PERK, CNX, SEL1L, Beclin1, P62, and LC3BII/LC3BI in duck cerebrums; increased the numbers of SEL1L and LC3B puncta in duck cerebrums. Thus, our data showed that excessive Cu could cause histopathological damage to duck cerebrums, disrupt the balance of the trace elements, induce oxidative stress and activation of ER quality control, thereby resulting in duck cerebrums damage.

Terbuthylazine (TBA) is one of the most commonly used and effective herbicides. However, due to its affinity for soil organic matter and water solubility, TBA can lead to biological health concerns. This study exposed broilers to TBA (0 mg/kg bw, 0.4 mg/kg bw, 4 mg/kg bw) for 28 days. The results showed significant pathological damage in broiler myocardial tissue, such as widening of the interstitial space, rupture of muscle fibers, and deposition of myocardial collagen fibers. In addition, Under the 0.4 mg/kg bw TBA exposure, myocardial oxidative stress was observed in broilers, which was accompanied by the activation of Nrf2/HO-1 pathway and the increased protein and mRNA levels of NQO1, NOX2 and SOD2 antioxidant enzymes. However, Nrf2/HO-1 protein and mRNA levels were reversed at 4 mg/kg bw TBA exposure. Meanwhile, the Nrf2/HO-1 mediated antioxidant defense was impaired. In contrast with the low dose, the protein and gene expression levels of NQO1, NOX2, and SOD2 were reduced in 4 mg/kg bw TBA group. The expression of GPX4 and SLC7A11 was significantly downregulated at both protein and mRNA levels. Beyond that, ACSL4 expression was significantly upregulated, and the protein result was consistent with the mRNA expression, demonstrating the occurrence of ferroptosis. In general, TBA exposure activated the Nrf2/HO-1 pathway, resulting in ferroptosis. This study links ferroptosis to the Nrf2/HO-1 pathway, providing new insights into the potential role of TBA in myocardial toxicity.

猫的股骨容易发生骨折,占猫骨折30％以上.本文的病例关于一只4岁雄性加菲猫因坠楼导致股骨骨折后的诊断和治疗措施.根据加菲猫的病症,先后对其由行视诊、触诊、血液生化指标检查,X射线检查,发现该猫左侧关节肿胀,丙氨酸氨基转移酶含量上升,左侧肋骨和股骨骨折.随后对其进行手术,术后用药护理.术后8天猫皮肤愈合良好,术后2月基本可以正常行走.

Copper (Cu) is an essential trace element for growth and development in most organisms. However, environmental exposure to high doses of Cu can damage multiple organs. To investigate the underlying mechanism of Cu toxicity on mitochondrial dynamics and mitophagy in the cerebrum of pigs, 60 30-day-old pigs were randomly divided into three groups and treated with different contents of anhydrous Cu sulfate in the diets (Cu 10 mg/kg, control group; Cu 125 mg/kg, group I; Cu 250 mg/kg, group II) for 80 days. The Cu levels and histological changes in the cerebrum were measured. Moreover, the protein and mRNA expression levels related to mitophagy and mitochondrial dynamics were determined. The results showed that the contents of Cu were increased in the cerebrum with increasing dietary Cu. Vacuolar degeneration was found in group I and group II compared to the control group. Additionally, the protein and mRNA expression levels of PINK1, Parkin, and Drp1 and the protein level of LC3-II were remarkably upregulated with increasing levels of dietary Cu. Nevertheless, the protein and mRNA expression levels of MFN1 and MFN2 and the mRNA expression of P62 were obviously downregulated in a Cu dose-dependent manner. Overall, these results suggested that excess Cu could trigger mitochondrial dynamics disorder and mitophagy in the pig cerebrum, which provided a novel insight into Cu-induced toxicology.

Type 1 diabetes mellitus (T1DM) is a chronic and represented by insulin-causing pancreatic β-cell disruption and hyperglycemia. N-Acetyl-Cysteine (NAC) is regarded as facilitating endothelial cell function and angiogenesis and may have treatment effect in the case of diabetes. However, the impact of NAC on T1DM are unknown. Here we reported that inflammatory pathogenesis of canine type 1 diabetes liver disease and the therapeutic effect of NAC combined with insulin. For this purpose, the model was established by intravenous injection of streptozotocin (20 mg/kg). Forty adult dogs were used and divided into 5 groups: control group, DM group, insulin treatment group, NAC combined with insulin therapy, and NAC group, while study lasted for 16 weeks. Results showed that the level of liver function enzyme activity were apparently increased in DM group, while the NAC with insulin treatment remarkable decreased liver function enzyme levels. Histopathology revealed that obvious changes in liver structure of all DM group, as evidenced by hepatocyte disorder and cellular swelling. Liver structure was evaluated by Periodic Acid Schiff (PAS) and Masson staining, the tissues appeared glycogen deposition and collagen deposition, indicating that DM aggravated liver injury. Compared with control group, the protein and mRNA expression of NLRP3, Caspase-1, ASC, and GSDMD were significantly induced in the DM group, while INS and NAC combined with INS treatment reversed the above changes. The levels of NF-κB P65, p-NF-κB, and IFN γ were availably enhanced in the DM group, which decreased through insulin and NAC combined with insulin treatment. This study demonstrated that NAC combined with INS exerted protective effects against STZ-induced liver injury by inhibiting the NLRP3/NF-κB pathway. The findings indicated that NAC combined with INS may serve as a potential candidate therapy for the treatment of T1DM.

Copper (Cu) is a common additive in food products, which poses a potential concern to animal and human health when it is in excess. Here, we investigated the relationship between endoplasmic reticulum (ER) stress and pyroptosis in Cu-induced toxicity of jejunum in vivo and in vitro. In in vivo experiments, excess intake of dietary Cu caused ER cavity expansion, elevated fluorescence signals of GRP78 and Caspase-1, and increased the mRNA and protein expression levels related to ER stress and pyroptosis in pig jejunal epithelium. Simultaneously, similar effects were observed in IPEC-J2 cells under excess Cu treatment. Importantly, 4-phenylbutyric acid (ER stress inhibitor) and MKC-3946 (IRE1α inhibitor) significantly inhibited the ER stress-triggered IRE1α-XBP1 pathway, which also alleviated the Cu-induced pyroptosis in IPEC-J2 cells. In general, these results suggested that ER stress participated in regulating Cu-induced pyroptosis in jejunal epithelial cells via the IRE1α-XBP1 pathway, which provided a novel view into the toxicology of Cu.

Neurodegenerative diseases are one of the major complications of type 1 diabetes mellitus (T1DM). The effect of insulin monotherapy on controlling blood glucose and neurodegeneration associated with diabetes is unsatisfactory. It is revealed that oxidative stress is a key element in T1DM. Therefore, N-acetylcysteine (NAC) was used together with insulin to investigate the therapeutic effect on neuronal damage in T1DM in this study. A total of 40 beagles were randomly divided into 5 groups (control group, DM group, insulin monotherapy group, NAC combined with insulin group, and NAC monotherapy group) to explore the effects of NAC on alleviating the oxidative damage in cerebrum. Our results showed that the contents of H2O2, 8-OHdg and MDA were apparently increased in DM group, while DNA and lipid oxidative damage was alleviated by the treatment of NAC and insulin. Histopathology revealed the sparse of neurofibrils and vacuolar degeneration in DM group. Additionally, compared with the control group, the mRNA expression levels of HO-1, nqo1, GCLC and GSTM1 were significantly decreased in DM group, while the opposite trend could be shown under NAC combined with insulin treatment. Meanwhile, the tight junction proteins of ZO-1, occludin and Claudin-1 were up-regulated with the treatment of NAC combined with insulin. Additionally, NAC further alleviated oxidative damage by enhancing the activity of GSH, Trx and TrxR and reducing the activity of catalase, GSSG and Grx to maintain redox homeostasis. These results demonstrated that NAC combined with insulin exerted protective effects against T1DM-induced cerebral injury via maintaining cerebral redox homeostasis.

Type 1 diabetes mellitus (T1DM) is a metabolic disease characterized by insulin deficiency and often accompanied by hypercholesterolemia. NAC is an effective antioxidative drug, but its application in the treatment of diabetes is still rare. A total of forty beagles were randomly divided into five groups: control group, DM group, INS group, INS with NAC group, and NAC group. The experiment lasted for 120 days. Results revealed that biochemical criterion increased in the DM group, while the indicators significantly decreased on the INS combined with NAC treatment group. Moreover, the insulin released test demonstrated that the model of T1DM was successfully constructed. The result of B ultrasound of gallbladder showed remarkable cholestasis in DM group. The cholesterol metabolism-related enzyme activity (HMGCR and SQLE) was evidently increased in DM group, but decreased in INS and NAC group. The content of TG. LDL-c, and HDL-c in liver was detected by the kit, and it was found that the content of TG, LDL-c, and HDL-c in DM group were reduced. Histopathological observation revealed that the cholestasis of liver cells and hepatic cords were disordered in DM group, the symptoms were alleviated under INS and NAC treatment. Additionally, the protein and mRNA expression of HMGCR and LDLR were obviously increased in DM group, but down regulated in INS and NAC treatment group. Overall, the liver function injury and secondary hypercholesterolemia can be found in T1DM canines, and NAC can relieve cholesterol metabolism disorder in the treatment of canine T1DM.

Diabetic nephropathy (DN) is a major complication of type 1 diabetes mellitus, and hyperglycemia and hypertension are the main risk factors for the development of DN. N-Acetyl-Cysteine (NAC) has a variety of effects, interfering with the production and scavenging of free radicals and regulating the metabolic activity of tissue cells. However, the efficacy of NAC on DN treatment is unclear. Thus, this study investigated the protective mechanism of NAC combined with insulin on renal injury in dogs with DN. The forty dogs were selected and divided into control group, DM group, INS group, INS + NAC group and NAC group to establish the model for a trial period of 4 months. The results revealed that INS + NAC was effective in reducing and stabilizing blood glucose levels. Biochemical results showed that INS + NAC treatment significantly regulated the stability of UREA, CREA and fructosamine indicators. Meanwhile, histopathology staining showed significant glomerular wrinkling and fibrosis in the DM group, which could be reversed after INS + NAC treatment. In addition, INS + NAC could restore mitochondria homeostasis by upregulating the levels of mitochondrial fission (MFN1, MFN2 and OPA1) and inhibiting of mitochondrial fusion (DRP1, FIS1 and MFF) related indicators. Further studies revealed that INS + NAC regulated the expression levels of renal BNIP3, NIX and FUNDC1 in the DM group, thereby alleviating mitophagy. Collectively, these results suggested that NAC combined with insulin protects DN by regulating the mitochondrial dynamics and FUNDC1-mediated mitophagy.

Cu is a metallic element that widely spread over in the environment, which have raised wide concerns about the potential toxic effects and public health threat. The objective of this study aimed to investigate the impression of copper (Cu)-triggered toxicity on mitochondrial dynamic, oxidative stress, and unfolded protein response (UPRmt) in fundic gland of pigs. Weaned pigs were randomly distributed into three groups, fed with different Cu of 10 mg/kg (control group), 125 mg/kg (group I), and 250 mg/kg (group Ⅱ). The trial persisted for 80 days and the fundic gland tissues were collected for further researches. Moreover, the markers participated to mitochondrial dynamic, UPRmt，and oxidative stress in fundic gland were determined. Results revealed that vacuolar degeneration were observed in the treated groups contrast with control group, and the Cu level was boosted with the increasing intake of Cu. Besides that, the levels of CAT, TRX, H2O2, and G6PDH were reduced in group Ⅰ and group Ⅱ, the mRNA levels of NRF2, HO-1, SOD-1, CAT, SOD-2, GSR, GPX1, GPX4, and TRX in the treated groups were promoted contrast to control group. Furthermore, the protein expression of KEAP1 was dramatically decreased, and the protein expression of NRF2, TRX and HO-1 were markedly enhanced in group Ⅰ and Ⅱ at 80 days. Moreover, the mRNA and protein expression levels of MFN1, MFN2, and OPA1 down-regulated and protein level of DRP1 was increased with the adding levels of Cu. Nevertheless, the UPRmt-related mRNA levels of CLPP, HTRA-2, CHOP, HSP10, and HSP60 were enhanced dramatically in Cu treatment group compared with control group. In general, our current study demonstrated that excessive absorption of Cu in fundic gland were related with stimulating UPRmt, oxidative stress, and the NRF2 interceded antioxidant defense. These results could afford an updated evidence on molecular theory of Cu-invited toxicity.

将120只1日龄商品代白羽肉鸡随机分为2组:对照组(铜含量11mg/kg)、高铜组(铜含量330mg/kg).并于试验49 d,采集盲肠组织及其内容物,制作组织切片观察组织变化,原子吸收法检测盲肠内容物铜含量,16S rDNA高通量测序检测盲肠内容物中微生物菌群结构和多样性.结果 显示,与对照组相比,高铜组盲肠中铜含量极显著升高(P＜0.01),且盲肠组织切片可以看出,高铜组盲肠绒毛高度显著降低(P＜0.05),隐窝深度显著增高(P＜0.05),绒毛高度与隐窝深度比值极显著降低(P＜0.01).微生物测序共获得OTUs 14988个,归属于10个细菌门,12个细菌科以及95个细菌属,其中对照组与高铜组所测菌群中,厚壁菌门和拟杆菌门的细菌所占比例最大,分别为10.82％和21.00％,属水平上另枝菌属所占比例最大.与对照组相比,随着日粮中铜含量的增加,门水平上,所测菌落中柔膜菌门丰度显著升高(P＜0.05),而厚壁菌门丰度显著降低(P＜0.05);科水平上,红螺菌科丰度极显著升高(P＜0.01),肠杆菌科丰度显著升高(P＜0.05),而韦荣球菌科丰度显著降低(P＜0.05);属水平上,另枝菌属丰度显著升高(P＜0.05),而厌氧棍状菌属、瘤胃梭菌属丰度显著降低(P＜0.05).结果 表明,饲料中铜含量的增加可以很大程度上降低肉鸡盲肠生物多样性,为高铜的毒性机理和早期诊断提供了理论依据.