Discovery medicine最新文献

Background: Cervical cancer is a major concern in women's health. Investigating the biological behavior of cancer cells can help to understand the underlying pathogenesis and offer novel insights into disease management. Therefore, this study evaluated the effect of irisin on the biological behaviors of cervical cancer cells and elucidated its underlying mechanism.

Methods: Cell viability of Caski and HeLa cells under different irisin concentrations was examined using the cell counting kit-8 (CCK-8) assay. Cell proliferation and autophagy levels were assessed at protein and mRNA levels using Western blot (WB) and quantitative real-time polymerase chain reaction (qRT-PCR) analyses, respectively. Apoptosis was determined by assessing the levels of activated caspase-3, caspase-8, and caspase-9 using corresponding enzyme-linked immunosorbent assay (ELISA) kits. The impact of irisin on cell cycle and apoptosis rates was evaluated using flow cytometry analysis. However, the migratory capability of irisin-treated cells was assessed using the scratch healing assay. Furthermore, expression levels of matrix metalloproteinases (MMP2, MMP7, and MMP9) were determined using WB analysis, and the Transwell assay assessed the invasive potential of the cells. The impact of irisin on macrophage polarization was examined through CD86 and CD206 typing using flow cytometry, and macrophage polarization status was determined by detecting the levels of inflammatory cytokines (interleukin (IL)-6, IL-10) and tumor necrosis factor-α (TNF-α). Then, THP-1 cells were directly co-cultured with cervical cancer cells to detect their effect on their biological behavior with or without irisin treatment, aiming to explore the underlying mechanism.

Results: Irisin reduces cervical cancer cell viability and decreases the protein and mRNA expression levels of minichromosome maintenance complex component 2 (MCM2), antigen identified by monoclonal antibody Ki67 (Ki67), and proliferating cell nuclear antigen (PCNA) (p < 0.05) in a dose-dependent manner, resulting in G0/G1 cell cycle arrest (p < 0.05). Irisin suppresses the expression of autophagy-related proteins Beclin-1 and microtubule-associated protein 1 light chain 3 (LC3) (p < 0.05), increases the content of cleaved caspase-3, cleaved caspase-8, and cleaved caspase-9 (p < 0.05), and enhances the apoptosis rate (p < 0.05). Additionally, irisin suppresses cervical cancer cell migration and reduces the expression of MMP2, MMP7, and MMP9 proteins (p < 0.05). Furthermore, it increases the number of CD86-positive cells (p < 0.05) while increasing the content of M1-type cytokine IL-6 and TNF-α (p < 0.05) and reducing the levels of M2-type cytokines IL-10 (p < 0.05), thereby promoting M1 polarization. The altered polarization state affects the apoptosis rate (p < 0.05), invasiveness (p < 0.05), and a

Background: Spermine (SPM) is known to play a role in regulating cholesterol efflux of macrophages, which is a critical anti-atherosclerotic pathway, but the underlying mechanism remains elusive. The deficiency of caspase 2 (CASP2), a target of SPM, can induce autophagy, which promotes cholesterol efflux in atherosclerosis. Herein, we aimed to explore whether SPM could regulate CASP2-mediated autophagy in the cholesterol efflux of macrophages.

Methods: Human THP-1 monocytes were induced into macrophages. After transfection and treatment with SPM or autophagy inhibitor 3-methyladenine (3-MA), the cholesterol uptake of THP-1 cell-derived macrophages and the lipid accumulation were examined using Dil-oxidized low-density lipoprotein (Dil-oxLDL) uptake assay and Oil Red O staining, respectively. The cholesterol efflux was measured by means of [³H]-cholesterol detection. Quantification of CASP2 and factors related to autophagy and apoptosis was completed using Western blotting and quantitative reverse transcription polymerase chain reaction (qRT-PCR).

Results: SPM treatment boosted cholesterol efflux and prevented lipid accumulation of THP-1 cell-derived macrophages (p < 0.001), without inducing cholesterol uptake of THP-1 cell-derived macrophages (p > 0.05). SPM upregulated the expression of autophagy-related 5 (ATG5), light chain 3 (LC3) II/LC3 I, and B-cell lymphoma-2 (Bcl-2) while downregulating Sequestosome 1 (P62) and CASP2 levels (p < 0.01). Treatment with 3-MA reversed the effects of SPM on cholesterol efflux, lipid accumulation, and autophagy-related protein expression (p < 0.01). Separately, CASP2 overexpression offset the impacts of SPM on cholesterol efflux, lipid accumulation, and expressions of proteins related to autophagy and apoptosis (p < 0.05).

Conclusion: SPM promotes autophagy to enhance the cholesterol efflux of THP-1 cell-derived macrophages by downregulating CASP2 expression.

Background: Soman is a highly toxic organophosphorus nerve agent that can cause persistent neurotoxicity. However, the mechanism of soman-induced neurotoxicity has not been completely clarified. Ferroptosis, a novel type of programmed cell death, has been confirmed to precipitate neurotoxicity caused by organophosphorus flame retardant. Since studies on the relationship between ferroptosis and soman-induced neurotoxicity are lacking, the purpose of this research is to explore the mechanism of soman-induced neurotoxicity from the perspective of ferroptosis.

Methods: Institute of Cancer Research (ICR) mice were injected subcutaneously with low-dose soman (1/2 × median lethal dose (LD₅₀)) daily for seven consecutive days to evaluate the neurotoxic injury caused by soman through alterations in body weight, cholinesterase activity, and neuronal counting. The morphologic changes of neurons and levels of ferroptosis-related molecules, including glutamate, glutathione (GSH), system xc^- (xCT), glutathione peroxidase 4 (GPX4), and malondialdehyde (MDA), were evaluated to confirm the involvement of ferroptosis in soman-induced neurotoxicity. The ferroptosis inhibitor ferrostatin-1 (Fer-1, 5 mg/kg) was administered daily immediately following soman exposure to further explore the role of ferroptosis.

Results: Repeated exposure to low-dose soman caused weight loss and neuronal death, and reduced cholinesterase activity. Meanwhile, it induced morphological changes of neurons characteristic of ferroptosis, increased glutamate and MDA levels, decreased GSH, and downregulated xCT and GPX4. Conversely, Fer-1 treatment mitigated neurotoxic injury and reversed the changes in ferroptosis-related molecule levels.

Conclusion: These findings indicate that ferroptosis engages in the course of neurotoxicity caused by soman and exerts a harmful effect via the glutamate/xCT/GPX4 pathway. This study provides new potential therapeutic targets for countermeasures against soman exposure.

Background: Hyperbaric oxygen (HBO) therapy functions as a possible therapeutic option for traumatic brain injury (TBI). The aim of this study is to detect the mechanism of HBO on TBI.

Methods: Neurons and astrocytes isolated from healthy neonatal rat cortices were co-cultured, a TBI model was established, and cells were cultured under HBO conditions. Neuron/astrocyte viability and glutamate transporter-1 (GLT-1) expression in neuron/astrocyte co-cultures were assessed by immunofluorescence. Tumor necrosis factor (TNF)-α/tumor necrosis factor receptor 1 (TNFR1)/nitric oxide (NO)/neuronal nitric oxide synthase (nNOS)/interleukin (IL)-1β/inducible nitric oxide synthase (iNOS)/GLT-1 levels in neuron/astrocyte co-cultures were detected using quantitative real-time polymerase chain reaction (qRT-PCR), colorimetry, and western blotting. To identify the key role of the target gene TNF receptor 1 (TNFR1) in HBO therapy, TNFR1 was silenced or overexpressed. After transfection, the cellular functions and the levels of related factors were re-examined.

Results: HBO (2 atmospheric absolute (ATA) for 30/60 min) attenuated the effect of TBI-induced on decrease of neuronal viability, increase of astrocyte viability, up-regulation of TNF-α, IL-1β, NO, nNOS, iNOS, and TNFR1 levels, down-regulation of GLT-1 levels, and reduce of GLT-1-positive astrocytes in neuron/astrocyte co-cultures (p < 0.05). TNFR1 knockdown and HBO (2 ATA for 60 min) enhanced neuronal viability, decreased astrocyte viability, and down-regulated TNF-α, IL-1β, NO, nNOS, iNOS, and TNFR1 levels in TBI-induced neuron/astrocyte co-cultures (p < 0.01). TNFR1 overexpression reversed the above role of HBO in TBI-induced neuron/astrocyte co-cultures. HBO (2 ATA for 60 min) up-regulated GLT-1 levels in TBI-induced neuron/astrocyte co-cultures (p < 0.05).

Conclusions: HBO inhibits TNFR1 expression to down-regulate NO content in TBI in an in vitro model.

Background: Bronchopulmonary dysplasia (BPD) is a common respiratory disease in premature infants. Nesfatin-1 is considered for the treatment of BPD. This study aimed to explore the anti-inflammatory effect of nesfatin-1 in the treatment of BPD.

Methods: Hyperoxia-induced newborn rats and transfected primary type II alveolar epithelial cells (AECIIs) were used to evaluate nesfatin-1's efficacy in treating BPD. Lung damage was assessed by means of wet-dry ratio measurement, Hematoxylin and Eosin staining, Masson staining, Terminal deoxynucleotidyl transferase-mediated dUTP nick-end labeling (TUNEL) staining, and Western blotting. Interleukin 6 (Il-6), tumor necrosis factor alpha (Tnf-α), and interleukin 1β (Il-1β) levels were measured by Enzyme-linked immunosorbent assay (ELISA) and quantitative polymerase chain reaction (qPCR). Neutrophils in bronchoalveolar lavage fluid were counted. High mobility group box 1 (HMGB-1), Toll-like receptor 4 (TLR4), nuclear factor kappa-light-chain-enhancer of activated B cells p65 subunit (p65), and NOD-like receptor family pyrin domain containing 3 (NLRP3) expressions were analyzed using Western blotting, while NLRP3 expression was detected through immunohistochemistry. AECIIs' viability, apoptosis, and reactive oxygen species (ROS) levels were assessed using cell counting kit-8 (CCK8), flow cytometry, and immunofluorescence, respectively. An immunofluorescence approach was used to detect surfactant protein C and ROS levels.

Results: In vivo, nesfatin-1 treatment significantly reduced the lung wet-dry ratio, increased the body weight of rats, inhibited apoptosis, alleviated lung damage, decreased inflammation, and lowered neutrophil counts (p < 0.05). In vitro, nesfatin-1 enhanced cell viability, inhibited apoptosis, decreased ROS levels (p < 0.01) and decreased mRNA levels of Il-6, Tnf-α, and Il-1β (p < 0.05) while increasing Il-10 mRNA (p < 0.01). In in vivo and in vitro scenarios, nesfatin-1 inhibited HMGB-1, TLR4, p65, and NLRP3 protein expression (p < 0.05). In hyperoxia cells, Hmgb-1 silencing showed similar results to those of nesfatin-1 treatment, while Hmgb-1 overexpression antagonized the effects of nesfatin-1 treatment.

Conclusion: This study showed that nesfatin-1 reduces neutrophils and suppresses inflammation via the HMGB-1/TLR4/Nuclear Factor kappa-light-chain-enhancer of activated B cells (NF-κB)/NLRP3 pathway, suggesting its potential clinical application in the treatment of BPD.

Background: To study the hard tissue change after tooth extraction filled with demineralized bone matrix (DBM) collagen scaffold bound with growth factors.

Methods: In 15 Beagle dogs, 90 socket sites were prepared by bilateral extraction of the mandibular premolars. Sockets were divided into six groups as follows: Group A: Blank (without any material); Group B: filled with HealiAid® Collagen Wound Dressing (SA001020, Maxigen Biotech Inc.); Group C: DBM; Group D: DBM-collagen-binding domain (CBD)-basic fibroblast growth factor (bFGF) (0.05 μg/mL); Group E: DBM-CBD-bFGF (5 μg/mL); Group F: DBM-CBD-bFGF (5 μg/mL) + bone morphogenetic protein 2 (BMP₂) (300 μg/mL). The changes in width and height of the buccal and lingual bone plates were determined by Cone-Beam Computed Tomography (CBCT) scan. At the 1st, 2nd, 3rd months after surgery, a total of 15 dogs were euthanized for Micro-computed tomography (Micro-CT) scan and histological examinations.

Result: At 1 month postoperatively, the differences in buccal bone height (BBH) of Group D [0.34 (0.18, 1.02) mm] and Group F [0.12 (0.04, 1.38) mm] were significantly lower than that of Group E [2.07 (1.03, 2.46) mm] (Group D vs. Group E, p = 0.032; Group F vs. Group E, p = 0.047). In the horizontal direction, from 1 to 3 months postoperatively, the median horizontal width changes at 1 mm, 3 mm and 5 mm below the alveolar ridge of both Group D and Group F were generally smaller than those of Group E, but there was no significant difference between the groups (p > 0.05). No significant differences were found among the indices of bone micro-structure measured by Micro-CT scan (p > 0.05). However, the fibers in the new bone zone of Groups E and F exhibited a higher level of maturity, as observed by Masson's trichrome stain.

Conclusion: Compared with the blank group and the control group, the DBM group with the addition of growth factors showed good alveolar ridge preservation, reduced bone resorption, and promoted the bone maturation of the extraction socket.

Background: Sepsis-mediated acute lung injury (ALI) has a high mortality rate, and glycyrrhizic acid (GA) possesses diverse pharmacologic activities. Herein, we investigated the role of GA in attenuating sepsis-triggered ALI.

Methods: Septic ALI was induced by cecal ligation and puncture (CLP). Mice were assigned into 5 groups with varied treatments. Their time of death was recorded every 6 hours after surgery. The wet/dry (W/D) weight ratio of the lung was measured. Observation of lung tissues was conducted by hematoxylin and eosin (HE) staining. Protein concentration in bronchoalveolar lavage fluid (BALF), levels of inflammatory cytokines, and production of reactive oxygen species (ROS) were detected by bicinchoninic acid (BCA), enzyme-linked immunosorbent assay, and dihydroethidium (DHE) staining, respectively. Endoplasmic reticulum (ER) stress-related genes, heme oxygenase-1 (HO-1), Janus kinase 2 (JAK2), and signal transducer and activator of transcription 3 (STAT3) levels were quantified by western blot.

Results: GA remarkably elevated survival rate and mitigated lung injury (p < 0.05). CLP markedly increased the W/D weight ratio and BALF protein concentration, while GA did the opposite (p < 0.05). CLP promoted interleukin-1β (IL-1β), tumor necrosis factor α (TNF-α), interleukin 6 (IL-6), protein kinase R (PKR)-like endoplasmic reticulum kinase (PERK), phosphorylation of eIF2α (p-eIF2α), activating transcription factor 4 (ATF4), C/EBP homologous protein (CHOP), the phosphorylation level of JAK/STAT3, along with DHE intensity, while GA showed opposite effects (p < 0.01). Additionally, GA markedly enhanced the HO-1 level (p < 0.05).

Conclusion: GA holds promise for future improvements in treating sepsis-induced ALI.

Background: Ischemia-reperfusion injury poses a significant challenge in cardiac pathology, leading to myocardial cell damage and dysfunction. Berberine, a natural compound, has shown potential cardioprotective effects. This study aims to investigate the protective role of berberine in myocardial cells post-ischemia-reperfusion injury, focusing on its modulation of the Neurogenic locus notch homolog protein (Notch) signaling pathway.

Methods: Male rats were subjected to ischemia-reperfusion injury and treated with berberine. Triphenyltetrazolium chloride (TTC) and hematoxylin and eosin (HE) staining were used to assess myocardial tissue damage and structure. Gene expression of Neurogenic locus notch homolog protein 1 (Notch1), Hairy and enhancer of split-1 (Hes1), B-cell lymphoma 2 (Bcl-2), and Bcl-2-associated X protein (Bax) was analyzed using Quantitative Polymerase Chain Reaction (qPCR). Apoptosis rate in myocardial cells was evaluated using Terminal deoxynucleotidyl Transferase dUTP Nick End Labeling (TUNEL) staining. Protein expression of Bcl-2 and Bax was examined. Molecular docking and Microscale Thermophoresis (MST) validation were performed to investigate the interaction between berberine and Notch1.

Results: Berberine treatment was associated with effective protection of myocardial tissue post-ischemia-reperfusion, indicated by reduced infarct area (p < 0.05) and improvements in tissue structure. Coinciding with these observations, berberine also influenced the expression of Notch1, Hes1, Bcl-2, and Bax in myocardial tissue, alongside modulation of Bcl-2 and Bax protein levels (p < 0.05). Additionally, molecular studies revealed that berberine binds to Notch1, suggesting a potential interaction.

Conclusion: The study suggests an association between berberine's cardioprotective effects and Notch signaling pathway modulation in the context of myocardial ischemia-reperfusion injury. While berberine was found to bind Notch1 and affect gene and protein expression related to apoptosis, further research is necessary to determine whether these effects directly contribute to reduced apoptosis and enhanced cell survival.

Background: Puerarin is the major bioactive ingredient extracted from Pueraria lobata. Puerarin has an antitumor effect on many kinds of cancer. Accordingly, the aim of this study was to investigate the effect and mechanism of puerarin on thyroid cancer (TC) proliferation and apoptosis.

Methods: TC and normal thyroid cells experienced exposure to puerarin at 0, 10, 50, or 100 μg/mL. Impacts of short hairpin RNA of Kruppel-like factor 2 (shKLF2), KLF2 overexpression, and notch receptor 1 (NOTCH1) overexpression on the malignant biological phenotypes of TC cells were gauged by cell function experiments. Quantification of KLF2, NOTCH1, B-cell lymphoma-2 (Bcl-2), Bcl-2-associated X (Bax), and Cleaved caspase 3 was completed using quantitative real-time polymerase chain reaction (qRT-PCR) and Western blot analyses. KLF2 and NOTCH1 levels in TC were analyzed using the Encyclopedia of RNA Interactomes (ENCORI) project database. Through rescue experiments, whether the anti-tumor effect of puerarin on TC is realized via KLF2/NOTCH1 axis was dissected.

Results: Puerarin inhibited the malignant growth of TC cells by up-regulating KLF2, which was reversed by shKLF2. KLF2 was lowly expressed in TC. Notably, NOTCH1 was negatively regulated by KLF2 and was highly expressed in TC. NOTCH1 overexpression abrogated KLF2 overexpression-inhibited malignant growth of TC cells, which was manifested as increased cell proliferation and Bcl-2 as well as decreased cell apoptosis, Bax and Cleaved caspase 3.

Conclusion: Puerarin suppresses TC cell proliferation and apoptosis via the KLF2/NOTCH1 axis.