Histology and histopathology Vol.40,nº10 (2025)
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- PublicationOpen AccessGalangin alleviates gastric mucosal injury in rats with chronic atrophic gastritis by reducing ferroptosis(Universidad de Murcia, Departamento de Histología e Histopatología, 2025) Yang Tian; Lu Min; Jiang Weiqiang; Jin Dandan; Sun Meiling; Mao Hua; Han Huixia; Biología Celular e HistologíaObjective. Chronic atrophic gastritis (CAG) is a precancerous lesion and is the first stage in a multistep precancerous cascade that can lead to gastric adenocarcinoma. This study aimed to reveal the role and mechanism of galangin in CAG. Methods. Rats were intragastrically administered a mixture of 2% sodium salicylate and 30% alcohol, forced to exercise, and fasted irregularly to establish CAG models. To explore the efficacy of galangin on CAG rats, we used Hericium erinaceus (HE) and omeprazole (Ome) as controls. The degree of gastric mucosal injury was assessed by H&E staining and immunohistochemistry. Perls staining and western blot analysis were used to assess iron content and enrichment of ferroptosis-related proteins. Reactive oxygen species and mitochondrial superoxide in the mucosa were visualized by probes. The morphology of cells was examined by transmission electron microscopy. Results. Our data showed that galangin treatment alleviated gastric mucosal damage and reduced ferroptosis in CAG rats, manifested as decreased iron content, iron transporters and storage proteins, decreased ROS and mitochondrial superoxide, and partially restored cellular morphology. Of note, galangin at a high concentration had better treatment efficacy than HE but lower than Ome. Conclusions. This study demonstrated that galangin reduced gastric mucosal injury in CAG rats by inhibiting ferroptosis. These findings provide a theoretical basis for its clinical application and broaden its potential applications
- PublicationOpen AccessShensiqigui tablets alleviate bleomycin-induced pulmonary fibrosis by inhibiting the hedgehog/wnt-β-catenin pathway(Universidad de Murcia, Departamento de Histología e Histopatología, 2025) Ma Rui; Xie Yupeng; Dong Yuan; Yin Fan; Yang Jiong; Xu Fenghua; Biología Celular e HistologíaBackground. Pulmonary fibrosis (PF) is a refractory disease characterized by inflammation and fibrosis. Shensiqigui Tablets (SSQGT), a combination of Codonopsis pilosula, Astragalus, and Angelica, is a traditional Chinese medicine with anti-inflammatory and antioxidant properties. Therefore, SSQGT may be a potential therapeutic agent for managing PF. This study aimed to investigate the effects of SSQGT on PF and its potential mechanisms. Methods. This study established a mouse model of PF through a single intratracheal injection of bleomycin (BLM) and used a TGF-β1-induced HFL-1 cell model. The experiment included control, model (BLM/TGF-β1), and treatment groups (pirfenidone, compound Biejiaruangan tablet (BJRGT), low-dose SSQGT, medium-dose SSQGT, and high-dose SSQGT). Histopathological changes and collagen deposition in lung tissues were observed using Hematoxylin-Eosin (HE) and Masson staining. Inflammatory exudation in bronchoalveolar lavage fluid (BALF) was assessed using ELISA, including TNF-α, IL-1β, IL-6, and NO. Oxidative stress markers SOD, GSH, and Malondi-aldehyde (MDA) were measured using commercial kits. mRNA and protein expression levels in lung tissues and in vitro models, including α-SMA, vimentin, collagen I, caspase-3, TGF-β, and the Hedgehog/Wnt-β-catenin pathway, were evaluated using qRT-PCR and western blot analysis. Results. SSQGT significantly alleviated BLM-induced weight loss and lung injury in mice and reduced HYP levels and collagen deposition. Additionally, SSQGT improved oxidative stress markers (decreased MDA levels and increased SOD and GSH activity) and mitigated inflammatory responses (reduced TNF-α, IL-1β, IL-6, and NO levels) and (downregulated α-SMA, collagen I, caspase-3, and TGF-β). Further mechanistic analysis showed that SSQGT inhibited the Hedgehog/ Wnt-β-catenin pathway. Conclusion. SSQGT alleviates BLM- or TGF-β1-induced PF by reducing oxidative stress and inhibiting inflammation through the suppression of the Hedgehog/ Wnt-β-catenin pathway, suggesting its potential as a therapeutic agent for PF
- PublicationOpen AccessOxidative stress markers 8-OHdG, Trx-1, and Prx6 are expressed in seminoma and are connected to TXNDC2, 3, and 6 expression levels(Universidad de Murcia, Departamento de Histología e Histopatología, 2025) Rantala Inka; Teppo Hanna-Riikka; Händelin Jonas; Kemppainen Janette; Ollikainen Riina K.; Kuittinen Outi; Kuusisto Milla E.L.; Biología Celular e HistologíaTestis-specific thioredoxins (TXNDCs) and oxidative stress have not been studied before in seminoma. We studied the immunohistochemical (IHC) expression of 8-OHdG, Prx6, Trx-1, TXNDC2, 3, and 6 in 25 testicular seminoma and 24 control samples. We retrospectively collected patient details and treatments from hospital records and combined them with IHC results. In normal testis, IHC expression of 8-OHdG, Prx6, TXNDC2, and 3 was higher than in seminoma samples (p<0.001 each), and the absence of n8-OHdG (p=0.002) and nTXNDC2 expression (p=0.044) was clinically associated with elevated lactate dehydrogenase levels. On the contrary, Trx-1 and TXNDC6 were over-expressed in seminoma samples (p=0.035 and p<0.001, respectively), and TXNDC6 was negatively associated with age over 40 (p=0.036). Concordant with the IHC expression, PRDX6 mRNA levels were downregulated with a fold change of -2517.4 (p<0.001), and TXN mRNA levels were upregulated with a fold change of 11637.4 (p=0.008) in seminoma samples compared with healthy controls. The oxidative stress markers studied had a correlation between the nuclear and cytoplasmic localization, a mutual correlation in expression between different markers, and mutual protein-protein interactions according to STRING Enrichment analysis. Our results show that oxidative stress markers and their expression in seminoma differ from that in normal testis tissue. Trx-1 and cTXNDC6 are seemingly overexpressed in seminoma, which might indicate their relevance in seminoma biology
- PublicationOpen AccessAdvantages and limitations of 3,3',5,5'-tetramethylbenzidine for immunohistochemical staining(Universidad de Murcia, Departamento de Histología e Histopatología, 2025) Yu Chao; Liu Xiao; Zhao Peiyuan; Sun Zhidong; Song Yurong; Cao Yuan; Cheng Ming; Biología Celular e HistologíaIn this study, two chromogenic systems, horseradish peroxidase (HRP)-3,3’-diaminobenzidine (DAB) and HRP-3,3',5,5'-tetramethylbenzidine (TMB), were used to perform single-color and double-color immunohistochemical staining (sIHC and dIHC, respectively) on multiple antigens in four distinct tissue types. The chromogenic results of the HRP-TMB system exhibited a vibrant blue-green color, and the tissue localization and signal intensity were consistent with those of the HRP-DAB system. In addition, it demonstrated clear differentiation from the hematoxylin-stained nucleus, endogenous melanin, and brown chromogenic results of HRP-DAB. TMB staining in tissues that contain high endogenous pigment levels eliminates the need for melanin bleaching, thereby facilitating direct observation and potentially improving the detection speed and interpretation. TMB can also be used in combination with DAB for dIHC, thus allowing detection of the two markers on a single slide. However, the TMB staining results are not stable over the long term and require image storage using slide scanners, thereby limiting its application. Additionally, in dIHC, overlapping signals of the first marker may obscure the second marker, potentially leading to bias or false negatives. Therefore, careful consideration is required when designing dIHC detection systems. Based on the above, we propose that TMB is a valuable supplement to DAB for immunohistochemical staining and deserves further promotion and utilization. However, additional research is needed to improve the composition of TMB chromogenic reagents, prolong the longevity of staining results, and overcome current limitations
- PublicationOpen AccessWnt5a regulates the expression of developmental genes in the adult retina following optic nerve crush injury(Universidad de Murcia, Departamento de Histología e Histopatología, 2025) Albano Gabrielle A.; Parrales Paola E.; Hackam Abigail S.; Biología Celular e HistologíaCanonical and non-canonical Wnt signaling pathways are well-characterized regulators of retinal development. Wnt signaling also promotes neuro-protection and regeneration in adult tissues, including retinal ganglion cell (RGC) survival and axonal regrowth after optic nerve injury. However, it is unknown whether Wnt-dependent neuroprotection after injury in the adult CNS is associated with altered expression of developmental genes. Müller glia are a prominent radial glia type in the retina that play critical roles in retinal neuron protection, RGC neurite growth, and axon regeneration by acting through Wnt and other signaling pathways. We recently used mass spectrometry to characterize proteins secreted from Müller glia in response to Wnt signaling. In this study, we investigated whether the Wnt-induced Müller glia secretome includes proteins involved in development and whether their corresponding genes are regulated by Wnt5a during axonal regeneration in a mouse model of optic nerve crush (ONC) injury. Adult mice received intravitreal injections of Wnt5a or saline at the time of ONC injury, and then retina tissue was collected at early time points post-injury. The expression of candidate Wnt-regulated developmental genes and related proteins were characterized by qPCR and immunohistochemistry. Our findings revealed that Wnt5a downregulated the expression of specific developmental genes, including cilia-related genes Nphp4, INTU, and Jade1, as well as transcriptional regulators Pax6 and Tsc1, with time-dependent changes observed during axonal regrowth. Several of these genes were localized to RGCs and inner nuclear layer cells, suggesting direct effects in RGCs and contributions from Müller glia. These results demonstrate that specific developmental gene pathways are suppressed by Wnt5a in association with RGC survival and axon regrowth following injury. Therefore, this study adds to our knowledge of potential mechanisms of Wnt-mediated optic nerve regeneration and identifies new categories of putative regeneration-regulating genes for further study
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