In Vitro Cellular & Developmental Biology. Animal最新文献

Rheumatoid arthritis (RA) is a chronic autoimmune condition that leads to joint damage. Mesenchymal stem cells (MSCs) are being recognized as a promising treatment option because of their capacity to modulate immune responses. Their therapeutic effects are mediated by released extracellular vesicles (EVs) which contain microRNAs known to influence inflammatory processes. This research focused on the impact of bone marrow MSC (BM-MSC)-derived EVs overexpressing miR-10a on cytokine production in a mouse model of collagen-induced arthritis (CIA). miR-10a was overexpressed in MSCs derived from bone marrow using Transfectamin. EVs were then isolated from the culture media of both miR-control and miR-10a-modified MSCs. Immunizing mice established the CIA model with type II collagen, after which they received either miR-control or miR-10a-enriched MSC-EVs. The severity of arthritis was evaluated through joint swelling measurements, and the concentrations of pro-inflammatory cytokines (such as interleukin (IL)-17a, interferon (IFN)-γ, and tumor necrosis factor (TNF)-α) alongside anti-inflammatory cytokines (including transforming growth factor (TGF)-β, IL-10, and IL-4) in the joints and serum were assessed using real-time PCR and enzyme-linked immunosorbent assay (ELISA), respectively. Our results indicated that treatment with miR-10a MSC-EVs led to a notable decrease in arthritis severity and joint damage in CIA mice. Furthermore, these EVs were found to lower levels of pro-inflammatory cytokines while enhancing anti-inflammatory cytokines compared to those treated with miR-control MSC-EVs. This study highlights how enhancing miR-10a expression can improve the therapeutic efficacy of MSC-EVs by altering the cytokine environment in CIA models.

Gastric cancer is a significant global health concern due to its high morbidity and mortality. Gastric cancer-associated mesenchymal stem cells (GC-MSCs) significantly contribute to its progression, with circ_0024107 being notably elevated in these cells and essential for their tumor-promoting activities. However, the expression and function of this circRNA in gastric cancer cells as well as its upstream regulators remain unclear. qPCR was used to assess circ_0024107 expression levels. Gain- and loss-of-function experiments evaluated its roles. Transwell assays measured cell migration and invasion. KHDRBS3 was predicted and validated through database analysis and qPCR, and its effects on circ_0024107 were analyzed using qPCR and transwell assays. The expression and clinical implications of KHDRBS3 in gastric cancer were evaluated using the TCGA-STAD database. circ_0024107 expression was elevated in gastric cancer cells, where it promotes migration and invasion. GC-MSCs further enhanced these capabilities by upregulating circ_0024107. KHDRBS3 was identified and validated as a regulator of circ_0024107 expression in both gastric cancer cells and GC-MSCs. Knocking down KHDRBS3 significantly reduced circ_0024107 levels, hindering gastric cancer cell migration and invasion, and weakening the influence of GC-MSCs on tumor cells. KHDRBS3 was abnormally elevated in gastric cancer tissues and correlated with patients' poor prognosis. KHDRBS3-mediated upregulation of circ_0024107 in gastric cancer cells and GC-MSCs synergistically enhances gastric cancer progression. This elucidates novel molecular interactions between GC-MSCs and gastric cancer cells, thereby presenting a promising therapeutic target for effectively mitigating gastric cancer metastasis.

As a persistent osteoarticular degenerative condition, intervertebral disc deterioration (IDD) has been established as a principal causative element in lumbar spine discomfort development. The present investigation seeks to assess the protective effects of casticin against IDD progression and elucidate associated molecular pathways. The CCK8 kit was used to assess the cytotoxicity of casticin on rat nucleus pulposus cells (NPCs). Western blot assay, qRT-PCR, enzyme-linked immunosorbent assay, reactive oxygen species assay, and immunofluorescence were used to detect the expression levels of inflammatory mediators and ROS production between different groups. The nuclear translocation of NF-κB p65 and expression of Nrf2/HO-1 signal pathway in lipopolysaccharide (LPS)-induced NPCs were detected by confocal microscopy. Moreover, histological analysis was used to evaluate the degree of disc degeneration in rats. Casticin treatment inhibited the production of oxygen free radicals and inflammatory mediators induced by LPS, such as ROS, TNF-α, IL-1β, and PGE2. Not only that, we also found that casticin retained the content of type II collagen and aggrecan in NPCs and inhibited the expression of MMP-13 and ADAMTS-5. Moreover, casticin treatment activated the Nrf2/HO-1 signal axis and inhibited nuclear translocation of NF-κB p65 in LPS-exposed NPCs. Histological analysis found that the treatment of casticin in rat IDD models prevented the loss of notochordal cells and the disordered arrangement of fiber loops. Casticin inhibits LPS-stimulated oxidative stress, inflammatory response, and ECM degradation by activating the Nrf2/HO-1 signaling axis and indirectly blocking the NF-κB pathway.

Valvular interstitial cells (VICs) are integral to the progression of calcific aortic valve disease (CAVD). Dentin matrix protein-1 (DMP-1), a member of the Sibling family protein, is implicated in the calcification process. This study aims to investigate the role and mechanisms of DMP-1 in the osteogenic differentiation of VICs. Between April 2018 and December 2018, aortic valve tissues were collected from 14 patients undergoing aortic valve replacement or heart transplantation. DMP-1 expression was quantified in calcified valves versus normal controls. An in vitro model of VICs' osteogenic differentiation was established to study the regulatory mechanism of DMP-1 on valvular calcification using immunoblot, immunohistochemistry, immunofluorescence, etc. The expression of DMP-1 was significantly increased in the calcified aortic valves patients (P < 0.01). DMP-1, both short and long arginine-glycine-aspartic acid (RGD) peptides, induced the osteogenic differentiation in VICs, an effect that was inhibited by an integrin αvβ3 antagonist (P < 0.05). Furthermore, the expression levels of RAF, RAS, MEK, and phosphorylated ERK1/2 were significantly elevated in VICs upon stimulation of DMP-1 (P < 0.05). DMP-1 is involved in the progression of valvular calcification and promotes the osteogenic differentiation of VICs via the integrin αvβ3 receptor. The combination of DMP-1 and integrin αvβ3 via its RGD domain activates the MAPK signaling pathway, leading to enhanced osteogenic gene expression in VICs. Clinical trial number: not applicable.

The mesothelial-mesenchymal transition (MMT) of peritoneal mesothelial cells is a critical factor contributing to the progression of peritoneal fibrosis. This study aimed to explore the effect of cGAS-STING signaling pathway on the MMT process in peritoneal mesothelial cells. The expressions of cGAS, STING, α-SMA, and Vimentin in HMrSV5 cells treated with high glucose were analyzed using WB. Subsequently, si-cGAS and the cGAS inhibitor RU.521 were employed to intervene in HMrSV5 cells. qPCR was utilized to evaluate the expression levels of genes involved in the cGAS-STING signaling pathway (cGAS, STING, IRF3, TBK1) and MMT-related genes (E-cadherin, Vimentin, α-SMA, TGF-β1). The protein expressions of the cGAS-STING signaling pathway and MMT-related proteins were detected by WB. The invasive capacity of cells in each cell was assessed using a Transwell assay, and the levels of pro-inflammatory cytokines (IL-6, TNF-α) in the supernatants of each cell were measured by ELISA. In the present study, we found that the expressions of cGAS, p-STING/STING, p-IRF3/IRF3, and p-TBK1/TBK1 proteins were significantly upregulated in HG-treated HMrSV5 cells. Furthermore, the activation of the cGAS-STING signaling pathway could be effectively suppressed in HMrSV5 cells transfected with si-cGAS or treated with RU.521. Additionally, treatment with si-cGAS or RU.521 not only attenuated the invasive capacity of HMrSV5 cells but also decreased the levels of pro-inflammatory cytokines and inhibited the expression of MMT-related markers. Suppression of the cGAS-STING signaling pathway mitigates HG-induced MMT in the human peritoneal mesothelial cell line HMrSV5.

Can artificial intelligence match or even outperform experienced embryologists in embryo selection for IVF transfer? To explore this question, we conducted a retrospective cohort study of 82 IVF cycles comparing embryo selections by an embryologist and AI (iDAscore), analyzing pregnancy outcomes. Embryologist and AI agreed in 64.6% of cases. Pregnancy rates were 45.2% in concordant vs. 44.8% in discordant selections (p > 0.05). AI-based embryo selection aligns closely with human judgment and may aid IVF decision-making. Larger studies are needed to confirm clinical utility.

The present study aimed to (1) evaluate the effects of different concentrations of the polysaccharide extract of Cissus sicyoides (PE-Cs) during in vitro culture of preantral follicles included in goat ovarian tissue on (i) follicular morphology and activation, (ii) ovarian stromal density, (iii) follicular and oocyte diameters, (iv) antioxidant enzymes activity (SOD, CAT, and GPx), (v) quantification of MDA, thiol, and nitrite levels; as well as to (2) measure the total antioxidant capacity of the extract. The ovarian cortex fragments were cultured at 39 °C in a humidified atmosphere with 5% CO₂ for 6 d in alpha-modified minimum essential medium (αMEM) supplemented with insulin, transferrin, and selenium; hypoxanthine; glutamine; and bovine serum albumin, which was called αMEM⁺ alone or added of PE-Cs at 20, 40, or 80 µg/mL. At the end of the culture period, a reduction in the percentage of normal follicles in all treatments using PE-Cs compared to fresh control and αMEM⁺. Moreover, 80 µg/mL of PE-Cs reduced stromal density and follicular diameter, as well as ultrastructural changes were observed in ovarian tissue. On the other hand, a decrease in MDA levels was observed in all treatments cultured with PE-Cs, although its antioxidant capacity was proven. In summary, supplementation of the culture medium with PE-Cs induced in vitro follicular degeneration. Thus, more studies are needed to evaluate the effect of Cissus sicyoides in reproductive cells and culture systems.

S100 protein family members S100A8 and S100A9 function primarily as a heterodimer complex (S100A8/A9) in vivo. This complex has been implicated in various cancers, including gastric cancer (GC). Recent studies suggest that these proteins play significant roles in tumor progression, inflammation, and metastasis. However, the exact mechanisms by which S100A8/A9 contributes to GC pathogenesis remain unclear. This study investigates the role of S100A8/A9 and its receptor in GC. Immunohistochemical analysis was performed on GC tissue samples to assess the expression of the S100A8/A9 receptor melanoma cell adhesion molecule (MCAM). In vitro transwell migration and invasion assays were used to evaluate the motility and invasiveness of GC cells. Cell proliferation was assessed using a growth assay, and Western blotting (WB) was employed to examine downstream signaling pathways, including ERK and the transcription factor c-Jun, in response to S100A8/A9-MCAM interaction. S100A8/A9 stimulation enhanced both proliferation and migration through MCAM binding in GC cell lines. These cellular events were accompanied by ERK activation and c-Jun induction. Downregulation of MCAM suppressed both ERK phosphorylation and c-Jun expression, highlighting the importance of the S100A8/A9‒MCAM‒ERK‒c-Jun axis in promoting GC progression. These findings indicate that S100A8/A9 contributes to GC progression via MCAM, which activates the ERK‒c-Jun pathway. The S100A8/A9‒signaling axis may represent a novel therapeutic target in GC.