Journal of Molecular Histology最新文献

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.

Gastric cancer (GC) is a highly heterogeneous disease that requires highly accurate prognostic models. Machine learning is a powerful tool for identifying predictive biomarkers and developing prognostic models. Here, we aim to integrate bioinformatics and machine learning algorithms to construct a risk model to predict prognosis of GC patients. Transcriptome data and clinical information of GC patients were obtained from the Cancer Genome Atlas (TCGA) database. Microarray data (GSE84437 and GSE26253) were obtained from the Gene Expression Omnibus (GEO) database. Univariate Cox regression analysis was used to screen prognostic genes. The risk genes closely related to prognosis were screened by machine learning algorithms and the risk score was calculated. Kaplan-Meier survival curve, time-dependent receiver operating characteristic (ROC) curve, univariate and multivariate Cox regression analysis were used to verify the validity of the risk model. The protein expression of hub genes in GC tissues was evaluated by immunohistochemical staining. 7 hub genes (CGB5, FEM1A, MATN3, ZNF101, MARCKS, BRI3BP and APOD) were identified and correlated with GC prognosis. A high-precision risk model based on random survival forest (RSF) and generalized boosted regression modelling (GBM) was constructed using these 7 hub genes. The risk model has good predictive ability for GC patients’ prognosis, and the risk score could be used as an independent prognostic factor for GC. In addition, the protein expression levels of CGB5, MATN3, MARCKS and APOD in GC tissues were significantly higher than those in normal tissues, and correlated with the pathological characteristics of GC patients. The risk model composed of 7 hub genes can accurately evaluate the prognosis of GC patients, which may contribute to the precise and personalized treatment of GC patients.

The oxytocin receptor (OTR) is known to be expressed in various organs, including the adult reproductive system. However, its expression in developing gonads has not yet been reported. This study investigates OTR expression in the ovaries and testes during critical developmental stages, specifically embryonic day 13.5 (E13.5), embryonic day 18.5 (E18.5), and postnatal day 14 (P14). According to our findings, both developing ovaries and testes exhibited OTR expression, including in immature follicle and Sertoli cells. OTR-expressing cells increased in the developing ovary, while a decrease was observed in the postnatal testis. In addition, our results showed that a high dose folic acid (FA) diet affected the number of OTR-expressing cells in developing gonads, particularly during the later stages of embryogenesis. These findings suggest that OTR may play a role in gonadal development and that its expression is influenced by sex. Furthermore, changes in folic acid supplementation may interfere with this developmental process.

The specific hepatoprotective mechanism of Fermented Panax ginseng C.A. Meyer extract (FGE) has not yet been fully elucidated. In this study, an AAPH-induced liver injury model was established to investigate the hepatoprotective effects of FGE. The hepatoprotective effects of FGE may involve the mediation of the NRF2 pathway by rare ginsenosides such as 20(R)-Rh1, 20(S)-Rg2, Rg5, 20(R)-Rg2, and 20(S)-Rh1. The results showed that FGE effectively improved pathological damage and liver dysfunction, increased SOD and GPx expression in the liver, and inhibited excessive ROS generation both in vivo and in vitro. Mechanistically, FGE regulated the mRNA and protein expression of KEAP1, HO-1, and NQO1, while enhancing NRF2 protein expression and promoting its nuclear translocation. In conclusion, the NRF2 signaling pathway activated by FGE may inhibit AAPH-induced oxidative stress and alleviate the resulting liver injury.

Graphical abstract