Journal of Molecular Histology最新文献

Aims

Methotrexate (MTX), a commonly used chemotherapeutic and immunosuppressive agent, is known to induce significant ovarian toxicity through mechanisms involving oxidative stress, inflammation, and apoptosis. Vortioxetine (VTX), a novel antidepressant with proven neuroprotective and anti-inflammatory properties, has not yet been evaluated in the context of chemotherapy-induced gonadotoxicity. This study aimed to investigate the protective effects of VTX against MTX-induced ovarian injury in a rat model by employing comprehensive histopathological and immunohistochemical evaluations.

Methods and Results

Thirty-two adult female Wistar Albino rats (300–350 g) were randomly divided into four equal groups (n = 8): Control, MTX, MTX + VTX, and VTX. Ovarian damage was induced with a single intraperitoneal injection of MTX (20 mg/kg), while VTX was administered daily (10 mg/kg) by oral gavage for five days. Rats were sacrificed on day 5, and bilateral ovaries were collected. Histopathological evaluation included follicular degeneration, vascular congestion, hemorrhage, and inflammatory cell infiltration. Immunohistochemical analyses were performed for 8-Hydroxy-2'-deoxyguanosine (8-OHdG), nuclear factor kappa B (NF-κB), tumor necrosis factor-alpha (TNF-α), caspase 3 (Cas-3), and Anti-Müllerian hormone (AMH) to assess oxidative stress, inflammation, apoptosis, and ovarian reserve. MTX administration caused severe follicular atresia, hemorrhage, and dense neutrophil infiltration. Immunohistochemically, 8-OHdG, NF-κB, TNF-α, and Cas-3 expressions were significantly elevated, while AMH was markedly reduced. VTX co-treatment significantly attenuated histological damage and modulated the expression of all biomarkers, indicating potent protective effects. VTX alone did not induce deleterious changes.

Conclusion

VTX exhibits a robust protective effect against MTX-induced ovarian injury via suppression of oxidative stress, inflammatory response and apoptotic pathways, while simultaneously preserving ovarian reserve. These findings highlight a novel application for VTX in fertility preservation strategies during chemotherapeutic interventions.

Acute pancreatitis (AP) is a serious inflammatory condition of the pancreas often requiring hospitalisation and characterised by abdominal pain, nausea, and vomiting. Hyperin (HP), a natural flavonoid, possesses antioxidant and anti-inflammatory characteristics. However, the effect of HP on AP remains unclear. This study aimed to evaluate the protective effects of HP in a cerulein-induced AP model in rats, focusing on histopathological damage, inflammatory cytokines, and oxidative stress markers. All rats were randomly assigned into control (n = 8), AP (n = 8), and AP + HP (50 mg/kg) groups (n = 8). We created an AP rat model by intraperitoneal cerulein injections. We evaluated histopathologically pancreatic tissues using hematoxylin–eosin staining. NF-κB and TNF-α expressions were analysed using immunohistochemical method. Oxidative stress markers such as MDA, SOD, CAT, GPx as well as serum amylase and lipase levels were assessed using biochemical methods. IL-6 and IL-10 levels were measured using ELISA. HP treatment significantly reduced histological damage scores, including oedema, necrosis, and vacuolisation. Moreover, expression levels of NF-κB and TNF-α were markedly decreased in the AP + HP group. HP restored SOD activity and reduced MDA levels, indicating attenuated oxidative stress. HP decreased serum IL-6 and increased IL-10 levels, along with significant reductions in amylase and lipase. HP exerts both anti-inflammatory and antioxidant effects in cerulein-induced AP, primarily through NF-κB and TNF-α inhibition and SOD activation. These findings suggest that HP may be a promising natural compound for the adjunctive treatment of AP.

Background and Aims

Cancer cachexia is a paraneoplastic syndrome characterized by progressive muscle atrophy, which negatively impacts treatment efficacy, quality of life, and survival in individuals with cancer. Despite extensive research, no effective medical intervention has completely reversed cachexia, primarily due to an incomplete understanding of its pathogenesis. Toll-like receptor 4 (TLR4) plays an important role in inflammation and metabolic regulation. In this study, the role of TLR4 in muscle catabolism was investigated, with a focus on its regulation of the p38 mitogen-activated protein kinase (MAPK)-mediated activation of C/enhancer-binding protein beta (C/EBPβ) pathway.

Methods

TLR4 expression was silenced in C2C12 myotubes using specific small interfering RNAs (siRNAs). Conditioned medium derived from various cancer cell types was applied to C2C12 myotubes to simulate the tumor microenvironment. The pharmacological TLR4 inhibitor TAK-242 was administrated to C2C12 myotubes and C26 tumor-bearing mice to evaluate its effects on muscle atrophy. Western blot analysis and immunofluorescence microscopy were performed on C2C12 myotubes, while muscle tissues from C26 tumor-bearing mice, a model of cancer cachexia, were analyzed using western blot and histological examination.

Results

Exposure to conditioned medium from cachexia-associated cancer cell lines induced p38 MAPK–C/EBPβ in C2C12 myotubes, leading to upregulation of Ubr2 and Atrogin-1, myosin heavy chain degradation, and myotube atrophy. Silencing or inhibition of TLR4 using siRNA or TAK-242 prevented these catabolic effects in vitro. In C26 tumor-bearing mice, TAK-242 administration significantly attenuated cancer-associated muscle atrophy.

Conclusions

TLR4 plays a critical role in cancer-associated muscle atrophy through the p38β MAPK–C/EBPβ signaling pathway in both in vitro and in vivo models. Pharmacological inhibition of TLR4 with TAK-242 effectively attenuated muscle atrophy, highlighting its potential therapeutic value.

The study aimed to investigate the histological, biochemical, and molecular effects of ethanol extract of Ferula orientalis in rats that had undergone cecal ligation and puncture (CLP). We chose Ferula orientalis because of its antioxidant and antibacterial properties. We divided 30 female rats into five equal groups. Group 1: Sepsis group, Group 2: Sepsis + Ferula LD, Group 3: Sepsis + Ferula HD, Group 4: Sham group, Group 5: Control. The cecum was removed from the abdomen and ligated with 4/0 sutures. After that two holes were created in the distal cecum by using 16-gauge needle to the CLP model. Histopathological (H&E, TUNEL, PERIODIC ACID-SCHIFF), Biochemical (SOD,CAT,GSH,MDA), and molecular (TNF–α, p53) analyses were performed. According to H&E findings, neutrophil infiltration, thickening of the alveolar walls, and areas of inflammation were quite remarkable in the sepsis group. In the TUNEL staining, a high density of apoptotic cells was noted in the sepsis group, while a dose dependent decrease in a density apoptotic cells was observed depending on the dose increase when the Ferula ethanol extract was applied. Antioxidant enzyme (SOD, CAT, and GSH) activity increased in the Ferula orientalis sepsis groups compared to the sepsis group (p < 0.05). The high-dose Ferula group showed the highest enzyme activity but had the highest level of TNF–α gene expression (p < 0.05). This suggests that Ferula may trigger different immune responses depending on the dose. The ethanol extract of Ferula orientalis improved the CLP sepsis model in rats by decreasing oxidative stress, inflammation and cell apoptosis.

Background

Myocardial fibrosis has been found to accelerate heart dysfunction after myocardial infarction (MI). Butein is a chalcone compound possessing multiple biological properties. However, its effect on MI-induced myocardial fibrosis remains unclear.

Methods

A mouse MI model was established by left anterior descending ligation. Human cardiac fibroblasts (HCFs) were stimulated with TGF-β1 in vitro. Mouse cardiac function was evaluated by assessing EF and FS. Masson’s trichrome staining showed the fibrosis area in murine hearts. Western blotting evaluated protein levels of fibrosis markers and signaling-related markers. CCK-8, EdU, and Transwell assays were used to evaluate HCF proliferation and migration.

Results

Butein improved MI-induced cardiac dysfunction and reduced the fibrosis area in mice. Butein inactivated TGF-β1/Smad signaling in MI mice and TGF-β1-stimulated HCFs. Butein inhibited TGF-β1-induced proliferation, migration, and collagen synthesis in HCFs, which were similar to the effects of LY2109761, a pharmacological inhibitor of TGF-β signaling.

Conclusion

Butein mitigated MI development by inhibiting cardiac fibrosis and ECM deposition by inactivating the TGF-β1/Smad signaling pathway.

Nasopharyngeal carcinoma (NPC) is a prevalent malignancy in Southeast Asia, particularly in southern China. Salvigenin is a natural compound with anti-inflammatory, antioxidant, and anti-tumor properties. The purpose of this research is to explore how Salvigenin impacts the malignant characteristics and immune evasion of NPC cells, as well as to clarify the molecular mechanisms involved. The viability and invasion of NPC cells (HK-1 and C666-1) were assessed using CCK-8 assays and Transwell assays, respectively. The expressions of epithelial-mesenchymal transition (EMT) markers and proteins in AKT/NF-κB pathway were analyzed by Western blot. Additionally, the impact of Salvigenin on tumor growth and immune evasion was examined in a xenograft mouse model. Salvigenin significantly inhibited proliferation and invasion of NPC cells in a dose-dependent manner. Besides, Salvigenin reduced the expression of PD-L1, inhibited CD8 + T cell apoptosis, and increased IFN-γ secretion, indicating attenuation of immune escape. After administration of Salvianin, the IFN-γ⁺ subpopulation was increased, but the TIM-3⁺ subpopulation was significantly reduced, indicating that Salvianin can inhibit T cell depletion. Mechanistically, Salvigenin inhibited the activity of AKT/NF-κB pathway, as evidenced by decreased levels of p-AKT and p-NF-κB. In addition, Salvigenin reduced tumor growth and immune evasion in vivo. Salvigenin exerts anti-tumor effects in NPC by inhibiting proliferation, invasion, EMT, and immune evasion via inactivation of the AKT/NF-κB pathway. Our findings illustrate that Salvigenin has potential as a new treatment option for NPC.

Graphical abstract

Salvigenin inhibits NPC cell proliferation, invasion and immune evasionthrough AKT/ NF-κB signaling pathway.

Renal ischemia/reperfusion injury is a critical clinical problem caused by kidney and heart surgery and can lead to acute kidney injury (AKI). Bevacizumab is a humanized monoclonal antibody that binds to circulating soluble isoforms of VEGF-A, thereby inhibiting the activation of VEGF molecular pathways and eliciting antiangiogenic effects. This study assessed the nephroprotective potential of bevacizumab in a rat model of renal ischemia/reperfusion injury (I/R). Twenty-four Sprague–Dawley rats were allocated into four groups: Sham, I/R, I/R + normal saline, and I/R + bevacizumab. The sham group was subjected to laparotomy without I/R induction. The I/R, I/R + normal saline, and I/R + bevacizumab groups were subjected to 30 min of bilateral renal ischemia, followed by 24 h of reperfusion. The rats in the I/R + normal saline and I/R + bevacizumab groups were administered normal saline (vehicle for bevacizumab) and 0.1 mg/kg bevacizumab via intraperitoneal injection 60 min before ischemia, respectively. Renal damage markers (creatinine and KIM-1), inflammatory and oxidative markers (TNF-α, IL-1β, NF-κB, F8-isoprostane and SOD), and an apoptotic marker (caspase-3) were measured via ELISA. Nrf2 and MLKL were assessed by IHC, and RIPK1 and HO-1 were assessed by RT‒qPCR, in addition to histological examination and molecular docking. Compared with the sham group, the I/R and I/R + normal saline groups presented significant increases in creatinine, KIM-1, NF-κB, TNF-α, IL-1β, F8-isoprostane, caspase-3, RIPK1, and MLKL and a reduction in SOD. Compared with those in the sham group, the histological findings in the I/R and I/R + normal saline groups revealed notable structural damage. Conversely, bevacizumab significantly reduced renal damage, inflammatory marker levels, cellular death, and histopathological findings. In bevacizumab-treated rats, the nuclear translocation of Nrf2 and HO-1 increased. Moreover, molecular docking analysis revealed that bevacizumab interacted with Keap1. Bevacizumab has nephroprotective effects against renal IRI by diminishing inflammation, necroptosis, apoptosis, and necrosis through the activation of the Nrf2/HO-1 pathway and the inhibition of the RIPK1/MLKL pathway.

Postherpetic neuralgia (PHN) is a debilitating chronic pain condition characterized by allodynia and hyperalgesia. Shengtong Zhuyu decoction (STZYD), a traditional Chinese medicine formula used clinically for pain associated with arthralgia syndromes and blood stasis obstructing the collaterals, was investigated for its potential role and mechanism in alleviating PHN. PHN was induced in male Sprague-Dawley rats via intraperitoneal injection of resiniferatoxin (RTX). STZYD was administered intragastrically to RTX-treated rats for 14 consecutive days. Its chemical components were identified using UHPLC-MS/MS. Analgesic efficacy was assessed by measuring mechanical paw withdrawal threshold and thermal paw withdrawal latency. The expression level of TRPV1 (a nociceptor and heat sensor) was analyzed using western blotting and immunofluorescence staining. ELISA was used to measure the levels of proinflammatory cytokines (IL-6, IL-1β, and TNF-α). Apoptosis was detected via TUNEL staining and western blotting. Underlying molecular mechanisms were evaluated using western blotting and immunohistochemistry. STZYD alleviated increased mechanical withdrawal threshold and thermal withdrawal latency and reducing TRPV1 levels in RTX-induced PHN rats. STZYD downregulated the levels of proinflammatory cytokines and pro-apoptotic proteins, reduced the number of TUNEL-positive cells, and upregulated the levels of anti-apoptotic proteins in RTX-induced PHN rats. Moreover, STZYD downregulated cAMP, PKA, and BNDF protein levels and inhibited CREB phosphorylation in RTX-induced PHN rats. Pharmacological activation of PKA by 8-Br-cAMP counteracted the protective effect of SDZYD in PHN rats. STZYD ameliorates RTX-induced mechanical allodynia and thermal hyperalgesia in rats by suppressing inflammation and DRG cell apoptosis through the inhibition of the cAMP/PKA/p-CREB signaling axis.

Acrylamide (ACR), a prevalent dietary toxicant formed in thermally processed foods via the Maillard reaction, is known to cross the placental barrier. While ACR-induced reproductive and developmental toxicity has been reported, the protective role of melatonin (MTN) via modulation of the SHH/GLI3 signaling pathway remains unclear. Pregnant Balb/c mice were divided into three groups (n = 6/group): control (distilled water), ACR (50 mg/kg/d), and ACR (50 mg/kg/d) + MTN (10 mg/kg/d), treated orally from gestational day (GD) 3.5 to GD 13.5. Placentas and embryos were collected for analysis. Oxidative stress (MDA levels), VEGF expression, and SHH/GLI3 pathway activity were assessed using immunohistochemistry and qRT-PCR. ACR exposure induced significant embryotoxicity, manifested as a 34% reduction in fetal weight (1.60 ± 0.09 g vs. 1.88 ± 0.14 g in controls, p < 0.001) and a 46.2% reduction in fetal crown-rump length (0.7 ± 0.08 cm vs. 1.1 ± 0.1 cm, p <  0.001). MTN co-treatment significantly ameliorated these growth restrictions. IHC analysis revealed that ACR significantly reduced SHH protein expression in the embryonic intestine and liver (p < 0.01), while it increased GLI3 protein levels (p < 0.01). MTN effectively normalized the expression of both proteins. At the molecular level, ACR downregulated SHH expression (p <  0.001) and upregulated GLI3 (p <  0.01), which were reversed by MTN. ACR exposure significantly increased oxidative stress (105% increase in placental MDA, p <  0.001) and reduced placental VEGF expression by 69.3% (p <  0.0001), both of which were significantly mitigated by MTN co-treatment. These integrated findings demonstrate that MTN exerts potent antioxidative and cytoprotective effects by mitigating ACR-induced oxidative stress, restoring SHH/GLI3 protein and gene expression, preserving VEGF-mediated placental angiogenesis, and preventing morphological defects. Our results underscore MTN’s therapeutic potential in counteracting ACR-induced teratogenicity and supporting healthy organogenesis.

The role of circular RNAs (circRNAs) derived from exosomes of mesenchymal stem cells in acute lung injury (ALI) is poorly understood. This study aimed to determine whether exosomal circRNAs can influence ALI and to uncover the underlying mechanisms. Bone marrow mesenchymal stem cells (BM-MSCs) were pretreated under hypoxic or normoxic conditions, from which exosomes were extracted (normoxic BM-MSC-derived exosomes (Nor Exo) and hypoxic BM-MSC-derived exosomes (Hypo Exo). To assess their in vitro effects on ALI, lipopolysaccharide (LPS)-treated MLE-12 cells were incubated with these exosomes. An ALI mouse model was established through airway perfusion with LPS, and exosomes were administered via the tail vein to evaluate their in vivo effects. The results showed that blocking exosome production could reverse the protective effect of the BM-MSC supernatant on LPS-induced injury in vitro. The exosomes attenuated LPS-induced ALI, with Hypo Exo demonstrating a more pronounced therapeutic effect than Nor Exo, both in vitro and in vivo. Additionally, a higher level of circ-LRP6 was detected in Hypo Exo compared with that in Nor Exo. Mechanistically, circ-LRP6 was found to regulate Claudin 4 levels in the context of ALI. These findings provide new insights into the potential of exosomal circ-LRP6 as a treatment for ALI.