In Vitro Cellular & Developmental Biology. Animal最新文献

Accumulative evidences have indicated the interaction between cellular senescence and ferroptosis. This study intends to investigate the ferroptosis-related molecular markers in TNF-α-induced endothelial senescence. The microarray expression dataset (GSE195517) was used to identify the differently expressed ferroptosis-related genes (DEFRGs) through weighted gene co-expressed network analysis (WGCNA). GO and KEGG were performed to explore the biological function. Furthermore, hub genes were identified after protein-protein interaction (PPI) analysis and validated through real-time qPCR (RT-qPCR). Then, a drug-gene network was established to predict potential drugs for the hub genes. Seven DEFRGs were recognized in the TNF-α-induced HUVEC senescence. Moreover, four hub genes (PTGS2, TNFAIP3, CXCL2, and IL6 are upregulated) were identified by PPI analysis and validated by RT-qPCR. Further analysis exhibited that PTGS2 was subcellularly located in the plasma membrane. Furthermore, after aminosalicylic acid (ASA) was identified as ferroptosis inhibitor for targeting PTGS2 in senescent HUVECs, 5-ASA and 4-ASA were verified to alleviate TNF-α-induced HUVEC senescence through ferroptosis. PTGS2 might play a role in TNF-α-induced HUVEC senescence and ASA may be the potential drug for alleviating TNF-α-induced HUVEC senescence through ferroptosis.

Pharmacological studies have shown that Cedrol (CE) exhibits extensive biological activities, including anti-inflammatory and analgesic. Moreover, it can inhibit the NF-κB pathway and the expression of various associated proteins. This study aimed to investigate the role of CE in postmenopausal osteoporosis. The results showed that intragastric administration of CE (10 and 20 mg/kg) significantly improved the bone microstructure damage and increased bone mineral density, trabecular bone volume, and bone trabecular thickness in ovariectomized (OVX) rats (p < 0.05). CE treatment additionally made a well-organized arrangement of bone trabeculae and improved its thickness and density. Compared with the OVX group, the levels of tartrate-resistant acid phosphatase from 5b and C-terminal telopeptide of type I collagen were significantly reduced by 42.75% and 49.27% in the OVX + CE rats (p < 0.05). TRAP staining visually showed that the number of osteoclasts in the femur tissue of CE-treated rats was less than that of the OVX group. The expressions of nuclear factor of activated T-cells, cytoplasmic 1, acid phosphatase 5, and cathepsin K in OVX + CE rats were significantly decreased by 51.61%, 46.07%, and 50.34% compared to the OVX group (p < 0.01). In addition, CE intervention effectively reduced the phosphorylation levels of P65 and IκBα and inhibited the NF-κB signaling pathway. Meanwhile, CE diminished the number of multinucleated osteoclasts induced by receptor activator for nuclear factor-κB ligand and hindered cell fusion as well as nuclear translocation of osteoclast precursor cells P65. In conclusion, CE inhibits osteoclastogenesis by suppressing the activation of the NF-κB signaling pathway, thereby alleviating postmenopausal osteoporosis.

A-1008Cadmium Affects Expression of Several microRNAs That Regulate the Wnt-beta Catenin Pathway in the hFOB 1.19 Osteoblast Cell Line. MICHAEL J. FAY, Elisha Pendleton, and Nalini Chandar. Midwestern University, Downers Grove, IL 60515. Email: mfayxx@midwestern.eduCadmium, an environmental pollutant, is known to cause bone damage and osteoporosis in individuals exposed to it. While several different mechanisms have been reported for their effects on bone, we and others have focused on the Wnt-beta catenin pathway as it plays an important role in osteoblast differentiation. As several microRNAs (miRNAs) can influence this pathway, we first conducted a study to determine the effect of cadmium using a human microRNA array containing 2632 unique probes. We found over 100 different miRNAs to be significantly altered by exposure of osteoblasts to 0.2 μM cadmium chloride for 24 hours. Roughly half of these miRNAs showed a significant increase or decrease in expression after cadmium treatment. We validated the changes seen in the microarray with a subset of these miRNAs using Realtime PCR. To determine how cadmium affects osteoblast differentiation, we chose a set of miRNAs that are known to either positively or negatively affect osteoblast differentiation via the Wnt/beta catenin pathway. The expression of these miRNAs was analyzed after exposure of cells to an osteoblast differentiation promoting media for 1, 3 and 7 days. As expected in osteoblasts miRNAs 119a, 15b, 21a, 24, 146a and 335 were increased robustly with differentiation. Cadmium treatment either resulted in no significant response or a stunted response. In the case of miRNAs that are known to negatively impact osteoblast differentiation, we found miR23b to be reduced during differentiation while cadmium treatment increased it. In the case of miRNAs 30b and 30d the levels remained reduced in both treatment types. Overall, these results show that treatment with cadmium can also affect miRNAs and cause an inhibition to osteoblast differentiation by altering the Wnt/beta catenin pathway.

HSP70 exhibits neuroprotective, antioxidant, and anti-apoptotic properties, which are crucial in preventing spinal cord injury (SCI) induced by oxidative stress and apoptosis. In this study, we assessed the potential protective effects and underlying mechanisms of HSP70 on tert-butyl hydroperoxide (TBHP)-damaged PC12 cells in an in vitro model of SCI. To establish the model, PC12 cells were subjected to oxidative damage induced by TBHP, followed by overexpression of HSP70. Cell viability was assessed using the CCK8 kit, intracellular reactive oxygen species level was evaluated using a commercial kit, cell apoptosis was detected using the Annexin V-APC/7-ADD Apoptosis Detection Kit, and the oxidative stress level was determined using SOD and MDA assay kits. Western blot analysis was used to detect the expression levels of Bax, cleaved caspase-3, and Bcl-2 proteins. Furthermore, immunofluorescence staining and Western bolt were used to detect the expression levels of proteins associated with the Nrf2/HO-1 signaling pathway. We found that HSP70 overexpression reduced apoptosis and oxidative stress in TBHP-induced PC12 cells. Furthermore, it activated the Nrf2/HO-1 signaling pathway. In addition, the Nrf2 inhibitor ML385 attenuated the protective effects of HSP70 on TBHP-induced PC12 cells. In conclusion, HSP70 can partially alleviate TBHP-induced apoptosis and oxidative stress in PC12 cells by promoting the Nrf2/HO-1 signaling pathway.

Cervus nippon (sika deer) are widely distributed throughout eastern Asia. Deer possess a variety of antibodies against several zoonotic pathogens, indicating that they act as reservoir of zoonoses. In this study, we reported the characterization of cultured cells derived from sika deer and evaluated their susceptibility to arthropod-borne viruses to clarify their usefulness in virological studies. Cells derived from testicular tissue in Dulbecco's modified eagle medium with 16% fetal bovine serum started growing as primary cultured cells. The diploid cells consisted of 68 chromosomes, consistent with those of Japanese sika deer previously reported. The phylogenetic analysis showed the cells formed a robust clade with Japanese population of C. nippon, indicating that the cultured cells established in this study were originated from the Japanese sika deer. The cells immortalized by the simian virus 40 T-antigen were predominantly spindle-shaped cells exhibiting adhesive properties, and cultivated at 37°C and 5% CO₂, which are common culture conditions for many mammalian cell lines. Western blotting analysis indicated that the cultured cells were multiple types of cells that coexist, including at least epithelial, fibroblast, and also Leydig cells. We confirmed that the cells have susceptibility to several arboviruses distributed in Japan: Getah virus, Japanese encephalitis virus, Oz virus, and severe fever with thrombocytopenia syndrome virus, but not to Tarumiz tick virus. From these results, the cells contribute to clarify the role of sika deer as a reservoir of zoonoses in nature and deer-associated experimental research at the cellular and molecular levels.