Journal of Biochemical and Molecular Toxicology最新文献

A series of 2,4-disubstituted pyrimidine derivatives bearing 5-substituted-1,3,4 thidiazole were devised and synthesized based on the binding mode of the approved drug Osimertinib with the ATP competitive site of EGFR-L858R/T790M in order to increase selectivity towards double mutant EGFR and potent antitumor activity. Their cellular bioactivity and corresponding enzyme inhibition were studied, and it was revealed that several compounds had significant biological activity and selectivity when compared to the control compounds. One of the most promising compound 8, substantially suppressed the proliferation of H1975 cells and showed significant inhibition of double mutant EGFR-L858R/T790M TK with IC₅₀ values of 0.170 and 0.0064 µM, respectively. Molecular mechanic simulation provides structural evidence of selective kinase inhibitory activity. Density functional theory (DFT/B3LYP) methods with the 6-311G**++ basic basis set were used to compute the theoretical vibrational frequencies and optimal geometric parameters. In addition, MESPs analysis, HUMO and LUMO quantum parameters of the most active compound 8 were calculated, and the results were viewed.

Pulmonary hypertension is a progressive disease associated with remodeling of the pulmonary vasculature. Excessive proliferation and migration of pulmonary artery smooth muscle cells (PASMCs) play important roles in nicotine-induced vascular injury. Connexin 43 (Cx43) is involved in intracellular communication and regulation of the pulmonary vasculature. However, the role of Cx43 and the potential mechanisms in PASMCs proliferation and migration induced by nicotine remains not very clear. In this study, we used both in vitro and in vivo models to explore the crucial role of Cx43 in pulmonary artery remodeling in nicotine treatment Tagln-Cre; Cx43^+/+ and Cx43 heterozygous (Tagln-Cre; Cx43^flox/+) or Cx43 Homozygous (Tagln-Cre; Cx43^flox/flox) deletion mice, and further explore the mechanism. We found that nicotine exposure led to modifications in the morphology and ultrastructure of pulmonary arteries in Tagln-Cre; Cx43^+/+ mice. Nicotine increased the Cx43 expression of pulmonary arteries and promoted the proliferation and migration of PASMCs of Tagln-Cre; Cx43^+/+ mice in a concentration-dependent manner by promoting ERK1/2 phosphorylation. Compared with the Tagln-Cre; Cx43^+/+ mice, the Tagln-Cre; Cx43^flox/+ mice were protected against increased ERK1/2 phosphorylation induced by nicotine. These results demonstrated that the downregulation of Cx43 reduced nicotine-induced proliferation and migration of distal PASMCs by inhibiting ERK1/2 phosphorylation.

Berberine, an isoquinoline alkaloid derived from various medicinal plants, emerges as a potential therapeutic agent against diverse human diseases. It has particularly shown notable anticancer efficacy against breast, colorectal, lung, prostate, and liver cancer. Berberine results in inhibition of cancer cell proliferation, induction of apoptosis, and suppressing angiogenesis, positioning it as a versatile, multitargeted therapeutic tool against cancer. Notably, berberine enhances the effectiveness of conventional chemotherapeutic drugs, mitigating associated drug resistance. Mechanistically, it has been shown to exert its efficacy by targeting molecules like nuclear factor-kappa B (NF-κB), mitogen-activated protein kinases (MAPKs), and phosphoinositide 3-kinase (PI3K)/Akt, thereby inhibiting survival pathways and promoting apoptosis of cancer cells. Moreover, berberine influences the expression of tumor suppressor genes, curtails cancer cell migration and invasion, and modulates the tumour microenvironment. Despite promising preclinical evidence, further research is essential to comprehensively elucidate its mechanisms of action and evaluate its safety and efficacy in clinical settings. In the present review, we have highlighted the pharmacokinetics, biosynthesis, and recent research work done pertaining to berberine's strong anticancer activity. We have also emphasised on the research being done on nanoformulations of berberine, which aim to improve its stability and bioavailability.

Our previous research identified that lncRNA PVT1 is upregulated in patients with IA. However, the precise functions of PVT1 in IA remain unclear. We compared the levels of PVT1, caspase-3, caspase-1, and NLRP3 in normal and IA patients. The GEO database was utilized to evaluate the expression of KLF4 and NLRP3 in IA. In vitro, we constructed transfected plasmids for silencing (si) and overexpression (oe) of PVT1 and USP10. We assessed cell apoptosis and measured the levels of IL-18, IL-1β, NLRP3, ASC, GSDMD-N, cleaved-caspase-3, and cleaved-caspase-1. CHX chase, immunoprecipitation assays, and bioinformatics tools were employed to evaluate the interactions among PVT1, USP10, and KLF4. Significant differences were observed in the levels of PVT1, KLF4, NLRP3, caspase-3, caspase-1, and histological examinations between normal and IA patients. Compared to the oe-NC group, the levels of IL-18, IL-1β, NLRP3, ASC, GSDMD-N, cleaved-caspase-3, and cleaved-caspase-1 were significantly increased in the oe-PVT1 and oe-USP10 groups. The effect of oe-USP10 was completely inhibited in the oe-USP10+si-PVT1 group. The RIPseq database demonstrated that PVT1 interacts with both USP10 and KLF4. The CHX chase assay showed that KLF4 interacts with both USP10 and PVT1. The RIP assay confirmed the interaction between PVT1 and USP10. The Co-IP assay and UbiBrowser indicated that KLF4 interacts with USP10. The ChIP assay demonstrated that KLF4 interacts with NLRP3. PVT1 may play a role in the pathophysiology of IA by regulating the USP10/KLF4/NLRP3 axis-mediated pyroptosis of HCSMCs.

Increasing long noncoding RNAs (lncRNAs) have been found to participate in regulating the progression of colorectal cancer (CRC), which is a common gastrointestinal malignancy. Here, the specific role and mechanisms of lncRNA LINC00294 were investigated in CRC. The expression levels of LINC00294, miR-499a-5p, and La-related protein 4B (LARP4B) in CRC cells (HCT116 and SW620) and tissues were assessed by RT-qPCR. The viability, proliferation, cell cycle, and apoptosis of HCT116 and SW620 cells were determined by CCK-8, EdU, and flow cytometry assays. Protein levels of cell cycle and cell apoptosis markers were measured by western blot analysis. FISH assay was performed to evaluate the subcellular localization of LINC00294 in CRC cells. Luciferase reporter assay, RNA pull-down assay, as well as RIP assay verified the interactions between miR-499a-5p and LINC00294 (or LARP4B). The xenograft model was established in mice to investigate the function of LINC00294 in vivo. LINC00294 presented a decreased expression level in CRC cells and tissues. LINC00294 overexpression suppressed the cell proliferative capacity, promoted cell cycle arrest, and induced cell apoptosis in CRC HCT116 and SW620 cells. LINC00294 interacted with miR-499a-5p, and miR-499a-5p targeted LARP4B. MiR-499a-5p was upregulated while LARP4B was downregulated in CRC cells. In rescue assays, LARP4B knockdown reversed the inhibition of LINC00294 overexpression on malignant phenotypes of HCT116 and SW620 cells. In the xenograft model, LINC00294 overexpression inhibited tumor growth in vivo. LINC00294 exerted an antitumor function in CRC by forming a LINC00294/miR-499a-5p/LARP4B regulatory network.

Among the diabetes-related complications, diabetic nephropathy is the major complication and it leads to end-stage renal failure. In the current investigation, we examined the protection of brucine, an alkaloid extracted from the seeds of Strychnos nux vomica plant widely used in traditional Chinese and Indian medicines against diabetic-induced nephropathy. Male Wistar rats were divided into four groups: control, diabetic rats, diabetic-induced + 40 mg/kg brucine-treated rats, and diabetic-induced + 25 mg/kg metformin-treated rats. The 35 mg/kg streptozotocin (STZ) was injected into (i.p.) rats to induce diabetes, and then rats were post-treated with brucine and metformin for 45 days. The food intake, weight gain, fasting blood glucose, HbA1C, insulin, and HOMA-IR values were measured on initial and after completion of treatment to confirm the induction of diabetes by STZ and the antidiabetic effect of brucine. The rats were euthanized on 45th day, and kidney tissue was isolated to assess the nephroprotective effect of brucine. The antioxidant property of brucine was analyzed by measuring the oxidative stress and antioxidant biomarkers. The lipid profiles in the control and treated rats were measured. To assess the nephroprotective effect of brucine KIM-1 protein was measured. The anti-inflammatory property of brucine was assessed by quantifying the levels of pro-inflammatory cytokines i.e., NF-κB, TNF-α, IL-1β, IL-6, TGF-β and to confirm the nephroprotective effect. The histopathological analysis was also performed with H&E staining. Our results suggested that brucine showed potent antidiabetic, antioxidant, antilipidemic, anti-inflammatory, and nephroprotective properties.

The underlying regulating mechanisms of miR-105-5p/PTEN in colon cancer (CC) progression are still unknown. MiR-105-5p and PTEN expressions were determined using RT-PCR. PTEN protein levels were examined by western blot. Also, the contents of inflammatory cytokines (IL–1β, TNF–α, and IL-6) were measured via ELISA. An inflammation model was constructed via LPS. NF–κB p65 expression was assessed via immunofluorescence assay. A xenograft tumor model was constructed in BALB/c nude mice. The functions of miR-105-5p were investigated by establishing a xenograft tumor model, H&E staining, TUNEL assay, immunohistochemistry assay, ELISA, RT-PCR, and western blot. In this research, We found that miR-105-5p expressions were upregulated in CC cells. MiR-105-5p depletion notably augmented PTEN expressions and enhanced immune response, while impeded EMT and distinctly declined the levels of p-IκBα and Nuc-NF-κB p65 in LoVo cells. Whereas, these effects were notably counteracted by PTEN depletion. MiR-105-5p upregulation exerted the opposite effects in CaCo2 cells. LPS markedly increased miR-105-5p expressions and suppressed PTEN expressions in LoVo cells. MiR-105-5p depletion offset LPS-triggered promoting effects on EMT and suppressing effects on the immune response. Meanwhile, in vivo assay proved that miR-105-5p depletion markedly impeded tumor growth and EMT, yet facilitated apoptosis and immune response. It also distinctly deactivated the NF-κB pathway. To sum up, these data indicated that miR-105-5p depletion might impede EMT, yet enhance the immune response in CC by elevating PTEN expressions via deactivation of the NF-κB pathway.

The aim of this study was to investigate the potential role of thymoquinone in the treatment of inflammatory bowel disease (IBD) by examining the effects of various doses of thymoquinone on histopathological changes, oxidative stress, and antioxidant markers in basic stamens in a 2,4,6-trinitrobenzene sulfonic acid (TNBS)-induced colitis model in rats. Thirty-two rats were divided into four groups: control, TNBS, thymoquinone-20 (20 mg/kg), and thymoquinone-50 (50 mg/kg) groups. The basic stamens of 32 rats were used for this experiment. Malondialdehyde, myeloperoxidase, and superoxide dismutase (inhibition rate) levels and histopathological scores (cellular infiltration, tissue integrity disruption, hemorrhagic focus, and mucosal loss) increased in the rats with TNBS, while they decreased with different thymoquinone doses. Glutathione peroxidase levels were decreased in the rats with TNBS, whereas increased with different thymoquinone doses. Furthermore, catalase levels were decreased in the rats with TNBS and increased with a 50 mg/kg thymoquinone dose. Malondialdehyde, catalase, myeloperoxidase, and superoxide dismutase (inhibition rate) levels and cell infiltration, tissue integrity disruption, and hemorrhagic focus scores detected in the thymoquinone-50 group were not different from the healthy control group. In conclusion, thymoquinone may be considered as a potential therapeutic agent in the treatment of IBD.

Sevoflurane (Sev) has a cardioprotective role in myocardial ischemia/reperfusion injury (MI/RI), but its mechanism has not been fully elucidated. This study aimed to investigate whether the circ_CDR1as/miR-671-5p/HMGA1 axis mediates the cardioprotective effect of Sev in MI/RI. Cardiomyocytes underwent hypoxia/reoxygenation (H/R) treatment was used to simulate MI/RI in vitro. H/R cardiomyocytes were then pretreated with Sev to explore the protective effect of Sev on H/R cells. The level of CDR1as/miR-671-5p/HMGA1 axis were detected by RT-qPCR. The proliferation and apoptosis of cardiomyocytes were detected by CCK-8 and flow cytometry. The levels of myocardial injury markers and inflammatory markers were detected by ELISA assay. Finally, the regulatory relationship between CDR1as and miR-671-5p/HMGA1 axis was verified by Dual-luciferase reporting and RNA pull-down assays. Sev Pretreatment can reduce the level of CDR1as and mitigate H/R-induced damage to cardiomyocytes. This Pretreatment lowers the levels of myocardial injury markers, oxidative stress markers, and pro-inflammatory factors in H/R-affected cardiomyocytes. However, CDR1as overexpression inhibits Sev's protective effect on H/R cardiomyocytes. At the molecular mechanism, we found that CDR1as mediates Sev's protective effect through the CDR1as/miR-671-5p/HMGA1 axis. CDR1as increases HMGA1 levels by sponging miR-671-5p, while high HMGA1 levels diminish Sev's protective effect. Sev plays a cardioprotective role in MI/RI by inhibiting the circ_CDR1as/miR-671-5p/HMGA1 axis.

Exposure to potassium dichromate (K₂Cr₂O₇) is well known for its nephrotoxic effects on humans and animals. This study investigated the protective effects of vitamin C against K₂Cr₂O₇-induced nephrotoxicity, focusing on its impact on altered carbohydrate metabolism, mitochondrial dysfunction, and associated molecular mechanisms in the cortical and medullary kidney segments. Male Wistar rats (n = 8) were divided into four groups: Group I received saline, Group II received a single 250 mg/kg body weight (bwt) intraperitoneal (i.p.) injection of vitamin C, Group III received K₂Cr₂O₇ (15 mg/kg bwt, i.p.), and Group IV received vitamin C 6 h before K₂Cr₂O₇ administration. Vitamin C significantly mitigated K₂Cr₂O₇-induced nephrotoxic effects, restoring normal renal function and histological architecture. It preserved the activities of glycolytic and gluconeogenic enzymes altered by K₂Cr₂O₇. Additionally, vitamin C mitigated K₂Cr₂O₇-induced mitochondrial dysfunction by maintaining tricarboxylic acid (TCA) cycle enzymes, electron transport chain proteins, mitochondrial DNA copy number, and ATP content. It also reduced oxidative stress markers and enhanced antioxidant enzyme activity. The protective mechanism of vitamin C against K₂Cr₂O₇-induced renal damage involved upregulation of the protein expression of peroxisome proliferation-activated receptor-γ coactivator-1α (PGC-1α), which further elevated the protein expression of nuclear factor erythroid 2-related factor-2 (Nrf-2) and transcription factor A, mitochondrial (TFAM), crucial for protecting cells from oxidative stress, enhancing mitochondrial function, and promoting cellular health. Overall, this study highlights the significant protective role of vitamin C against K₂Cr₂O₇-induced renal damage by preserving carbohydrate metabolism and mitigating mitochondrial dysfunction through the PGC-1α/Nrf-2/TFAM pathway, offering valuable insights into its protective mechanisms in nephrotoxicity.