In Vitro Cellular & Developmental Biology. Animal最新文献

Channel catfish virus (CCV) poses a significant threat to catfish culture. Lack of effective vaccines and antiviral treatments necessitates effective disinfection strategies to mitigate its spread. In vitro trials indicated the virus to be inactivated at high temperatures, but was infectious at 40°C. This study evaluated the efficacy of a commercial disinfectant against CCV using both in vitro and in vivo approaches. In vitro experiments assessed the virucidal activity of the disinfectant against CCV in channel catfish ovary (CCO) cells, while in vivo trials evaluated its effectiveness in reducing viral transmission and mortality among channel and hybrid catfish fingerlings. Results indicated that the disinfectant was effective in inactivating the virus at the tested concentrations and improved the survival of fish exposed to the virus. This study provides critical insights into selecting appropriate disinfection protocols to enhance biosecurity in catfish hatchery settings and to mitigate CCV transmission.

Osteoarthritis (OA) is a chronic degenerative disease characterized by the progressive loss of articular cartilage. The role of cigarette smoke (CS) in OA is debated, with some studies suggesting a protective effect while others indicate it may pose a risk. Our preliminary findings suggest a link between smoking in young adults and severe knee OA, though the extent of this contribution is unclear. This study investigates the impact of cigarette smoke extract (CSE) on human chondrocytes. Human chondrocyte cultures were exposed to varying concentrations (0-10%) of CSE for 7 d. We evaluated cell viability, extracellular matrix (ECM) components, metalloproteinase expression and cytokines levels, and antioxidant enzymes (SOD1 and CAT) using calcein staining, immunohistochemistry and ELISA. Oxidative stress (OS) was assessed by measuring hydrogen peroxide (H₂O₂) and nitric oxide (NO) levels. Results were analyzed using ANOVA with Tukey post hoc tests, and Pearson correlation coefficients were calculated. Cell viability decreased at 10% CSE, and ECM components were diminished. MMP9 and MMP13 expression significantly increased at 5% and 10% CSE. H₂O₂ levels peaked at 1%, while IL-1β peaked at 2.5%. Antioxidant expression (SOD1 and CAT) decreased at higher concentrations, and heat shock protein 70 (HSP70) was notably expressed. MMPs expression was negatively correlated with both viability and ECM components. CSE induces cellular damage, alters ECM composition, and upregulates MMP expression via OS and IL-1β, while diminishing antioxidant defenses. These findings suggest that smoking may disrupt articular cartilage homeostasis, highlighting the need for further investigation into oxidative stress and inflammatory mediators.

This study aimed to investigate the expression, prognostic significance, methylation, and immune invasion levels of secreted frizzled-related proteins (SFRP1-5) in colorectal cancer (CRC). Additionally, the relationship between SFRP1/2 methylation and immune infiltration in CRC was explored. The expression of SFRP1-5 was analyzed using several databases, including GEO, TCGA, TIMER, STRING, and GEPIA. Molecular interactions with SFRPs were examined via Cytoscape software. Gene Ontology (GO) and Kyoto Encyclopedia of Genes, and Genomes (KEGG) pathway analyses were conducted using the DAVID database. Methylation levels of SFRP1/2 in CRC were assessed through methylation-specific PCR (MSP) and bisulfite sequencing PCR (BSP) experiments. Apoptosis and proliferation in CRC cells following the knockdown of SFRP1/2 expression were evaluated using flow cytometry and CCK-8 assays. The TISIDB database was used to analyze the relationship between SFRP1/2 methylation levels and immune infiltration. The expression of SFRP1, SFRP2, and SFRP5 was significantly lower in CRC patients, while SFRP4 expression was higher compared to that in healthy individuals. Elevated mRNA expression of SFRP2 was significantly associated with improved overall survival (OS), disease-specific survival, and progression-free intervals. SFRP1/2 expression was also linked to immune invasion, with higher levels correlating with increased immune infiltration. Both SFRP1 and SFRP2 showed hypermethylation in CRC. Knockdown of SFRP1/2 expression resulted in increased proliferation of CRC cells, and their methylation levels were inversely correlated with immune cell presence. The expression, methylation, and immune cell infiltration patterns of the SFRP family in CRC differed markedly from those in healthy individuals. These findings suggest that SFRPs may serve as potential therapeutic targets and key genes associated with immune cell infiltration in CRC.

Colorectal cancer (CRC) is an extremely harmful malignant tumor. Optic atrophy 3 (OPA3) is highly expressed in multiple tumors, but its action in CRC is still unknown. This research aims to explore the role of OPA3 and its related molecular mechanisms for CRC. Firstly, we overexpressed and knocked down OPA3 to examine its effect on CRC cell (HT29 cell) growth. CRC cell viability, migration, invasion, and levels of proliferation markers and cell cycle-associated proteins were measured. Then, we treated cells with carbonyl cyanide m-chlorophenyl hydrazone (CCCP) to explore mitochondrial dysfunction and mtDNA stress in HT29 cells. Next, we overexpressed cGAS and STING to examine their correlation with OPA3. The results showed that OPA3 overexpression enhanced CRC cell viability, migration, invasion, and the levels of PCNA, Cyclin A2, and Cyclin B1. Knockdown of OPA3 had the opposite effects. Moreover, OPA3 knockdown facilitated mitochondrial dysfunction and mtDNA stress in CRC cells. OPA3 overexpression also inhibited CCCP-induced mitochondrial stress disorder. Additionally, OPA3 knockdown elevated the protein levels of p-STING and cGAS and the mRNA level of STING target genes. Furthermore, overexpression of either cGAS or STING partially alleviated the enhancement of HT29 cell proliferation, migration, and invasion mediated by OPA3 overexpression. In conclusion, OPA3 promotes CRC progression via inhibiting the cGAS-STING pathway, which is mediated by mtDNA stress. OPA3 may be a new potential target for CRC.

Maxing Yigan formula (MYF) is a traditional Chinese medicine (TCM) prescription used for the treatment of OA for decades in China. However, the mechanism remains unknown. In this study, we developed a MYF-incorporated collagen sponge (MYF@CS) and investigated its cartilage regeneration effect and the underlying mechanism. In vitro experiments revealed that MYF significantly promoted cell viability, proliferation, and autophagy of OA chondrocytes. Furthermore, MYF@CS significantly enhanced chondrogenesis and cartilage regeneration, as assessed by macroscopic observation, the International Cartilage Repair Society (ICRS) visual histological score, and histological examination. Our findings suggest that MYF@CS could represent a significant therapeutic strategy for the treatment of OA.

The expression of matrix metalloproteinase (MMP)-7 is reported to be correlated with invasion and metastasis of colorectal cancer (CRC). Therefore, the inhibition of MMP-7 would be beneficial for the suppression or prevention of CRC cell invasion and metastasis. The stem bark of Acanthopanax senticosus, a widely used medicinal herb, contains isofraxidin (IF) and chlorogenic acid (CGA) as major components. Previously we reported that IF suppressed the expression of MMP-7 and cell invasion in human hepatoma cells. In this study, we investigated the effects of CGA on cell invasion, MMP-7 mRNA expression and the phosphorylation of extracellular signal-regulated kinase 1/2 (ERK1/2) and compared it with those of IF in human CRC cells (HT-29). We found that CGA significantly suppressed 12-O-tetradecanoylphorbol-13-acetate (TPA)-induced cell invasion, MMP-7 expression and the expression of activated form of MMP-7 to almost the same extent as IF. Meanwhile, we also found that TPA-induced expression of MMP-7 mRNA and ERK1/2 phosphorylation were significantly suppressed when cells were exposed to the ERK1/2 inhibitor U0126 and that CGA was a little more potent than IF at inhibiting TPA-induced ERK1/2 phosphorylation. Taken together, the present results indicate that CGA suppresses cell invasion, MMP-7 expression and ERK1/2 phosphorylation to almost the same extent as IF and suggest that not only IF but also CGA suppresses cell invasion by inhibiting MMP-7 expression via the inhibition of at least ERK1/2 phosphorylation and Acanthopanax senticosus, which contains two components with anti-MMP-7 activity, may be a beneficial herb with anti-invasive effects against human CRC cells.

Chitosan-based hydrogels have gained considerable attention in biomedical research due to their inherent biocompatibility, biodegradability, and non-toxicity. When combined with polyvinyl alcohol (PVA), the resulting hydrogels exhibit superior mechanical strength, elasticity, and swelling capacity, making them highly suitable for a range of applications, including wound healing, tissue engineering, and controlled drug delivery. In this study, we synthesized and characterized a novel PVA/gelatin/chitosan (PVA/G/C) hydrogel composite using a series of analytical techniques such as Fourier transform infrared spectroscopy (FTIR), X-ray diffraction (XRD), scanning electron microscopy (SEM), and energy-dispersive X-ray analysis (EDAX). The morphological, structural, and compositional analyses confirmed the successful formation of a homogenous, porous network conducive to cell proliferation and nutrient diffusion. In this study, polyvinyl alcohol/gelatin/chitosan-based hydrogels were prepared to study the potential for micro-tissue formation and wound healing application using Danio rerio gill (DrG) and Danio rerio fin (DrF) cell lines, respectively. Overall, the findings indicated the potential use of PVA/G/C hydrogel films as wound dressings. The idea of creating physically cross-linked hydrogels of PVA and chitosan without using any harmful organic chemicals or solvents is the novelty of this work. This study highlights the versatility and potential of PVA/G/C hydrogels, not only as a promising material for wound healing and drug delivery but also as an effective scaffold for tissue engineering applications.

MicroRNA-146a-5p (miR-146a-5p) actively participates in the process of cerebral ischemia-reperfusion (CI/R) injury. Dysregulation of the tumor necrosis factor receptor-associated factor 6 (TRAF6)/nuclear factor kappa-B (NF-κB) p65 axis is closely associated with inflammatory response. This study aimed to investigate the potential involvement of miR-146a-5p and TRAF6/NF-κB p65 in mediating CI/R progression in vitro. HT-22 cells were challenged with oxygen-glucose deprivation/reoxygenation (OGD/R) to simulate CI/R in vitro. HT-22 cells were transfected with miR-146a-5p mimics or TRAF6 overexpression constructs. The impact of miR-146a-5p on apoptosis, inflammation, and TRAF6/NF-κB p65 activation were investigated. OGD/R inhibited HT-22 cell viability, induced apoptosis, reduced miR-146a-5p levels and activated the TRAF6/NF-κB p65 pathway. MiR-146a-5p mimics reduced pro-inflammatory factor release, limited apoptosis-related protein expression, and inactivated the TRAF6/NF-κB p65 pathway in OGD/R-challenged HT-22 cells. Mechanistically, miR-146a-5p was verified to bind to TRAF6 3'UTR. TRAF6 overexpression reversed the beneficial effects of miR-146a-5p mimics on apoptosis, inflammation, and TRAF6/NF-κB p65 activation. This work revealed that miR-146a-5p targeted TRAF6 and suppressed the TRAF6/NF-κB p65 pathway, thereby reducing OGD/R-induced inflammation and apoptosis in HT-22 cells. These findings suggest the potential of the miR-146a-5p/TRAF6/NF-κB p65 axis in the treatment of CI/R.