In Vitro Cellular & Developmental Biology. Animal最新文献

Multiple sclerosis (MS) is a neurodegenerative and autoimmune disease affecting the central nervous system (CNS). Recently, mesenchymal stem cells (MSCs) and their extracellular vesicles (EVs) have been extensively studied as a potential treatment for MS. In this study, we examined the impact of therapy using EVs derived from murine bone marrow MSCs (BMSC-EVs) on the proliferation of splenocytes, frequency of regulatory T cells (Tregs), and cytokine secretion in mice induced with experimental autoimmune encephalomyelitis (EAE), comparing the effects with those of their parent cells. After inducing EAE in 30 mice, the animals were divided into three groups and treated with PBS, BMSCs, or BMSC-EVs. The mice were sacrificed on day 30 post-immunization, and their splenocytes were isolated for further analysis. The proliferation of splenocytes was assessed by measuring the fluorescent intensity of CFSE dye using a FACSCalibur flow cytometer, the frequency of Treg cells was determined by flow cytometry, and cytokine levels of tumor necrosis factor-alpha (TNF-α), interleukin (IL)-1β, IL-6, IL-10, and transforming growth factor-beta (TGF-β) were measured using enzyme-linked immunosorbent assay (ELISA). The results showed that treatment with BMSC and BMSC-EV both significantly reduced splenocyte proliferation, increased Treg cell frequency, and shifted cytokine profiles toward reduced pro-inflammatory (TNF-α, IL-1β, IL-6) and increased anti-inflammatory (IL-10, TGF-β) cytokines compared to untreated EAE controls, with comparable efficacy between BMSCs and BMSC-EVs. These findings emphasize the capability of BMSC-EVs to serve as a cell-free therapy for immune response modulation in EAE.

Recurrent spontaneous abortion (RSA) represents a substantial challenge in reproductive medicine, attributed to a variety of complex factors, among which aberrations in long non-coding RNAs (lncRNAs) play a crucial role. The present study delves into the functional dynamics of the lncRNA H19 in the context of RSA, particularly focusing on its regulatory interplay with miR-29a-3p and the Suppressor of Cytokine Signaling 3 (SOCS3). A notable downregulation of H19 in villous tissues from RSA patients was observed, highlighting its potential involvement in RSA pathophysiology. Functional assays demonstrated that overexpression of H19 in HTR-8/SVneo cells enhances cellular viability while concurrently attenuating apoptotic processes, thereby indicating a pivotal role of H19 in cellular survival pathways. This study identifies miR-29a-3p as a direct regulatory target of H19, exerting significant influence on cellular viability and apoptosis. The inhibition of miR-29a-3p was observed to mitigate its pro-apoptotic effects, thereby reinforcing its critical regulatory capacity in cellular homeostasis. Moreover, SOCS3 was delineated as a downstream effector of miR-29a-3p, with its expression being inversely modulated by miR-29a-3p. Co-transfection experiments involving H19, miR-29a-3p, and SOCS3 unraveled their intricate regulatory nexus in modulating cellular survival mechanisms. Collectively, these findings elucidate that H19 orchestrates the regulation of cell viability and apoptosis in RSA through the miR-29a-3p/SOCS3 signaling axis, thereby providing valuable insights into the molecular underpinnings of RSA and unveiling novel avenues for therapeutic intervention.

Effective neovascularization is critical for tissue repair and the enhancement of cardiac function following myocardial infarction (MI). However, the hypoxic microenvironment post-MI significantly impedes neovascular formation. Although ATF4 has been implicated in heart failure and myocardial cell regeneration and repair, its role in angiogenesis remains unclear. This study utilized both in vitro and in vivo models to investigate the role of ATF4 in neovascularization after MI. In hypoxia-cultured murine endothelial cells (ECs), hypoxia was observed to inhibit EC proliferation, migration, and tube formation. In contrast, overexpression of ATF4 ameliorated these hypoxia-induced impairments. Conversely, inhibition of ATF4 further exacerbated the reduction in EC proliferation, migration, and tube formation induced by hypoxia. Notably, the beneficial effects of ATF4 were reversed by the PI3K/AKT inhibitor LY294002. Under hypoxic conditions, ATF4 overexpression significantly upregulated phosphorylated (p)-PI3K, p-AKT (T308), and p-AKT (S473) in ECs. LY294002, however, markedly reduced the expression of p-PI3K, p-AKT (T308), and p-AKT (S473) in hypoxic ECs overexpressing ATF4. In a murine MI model, ATF4 overexpression partially mitigated cardiac dysfunction and promoted neovascularization, effects that were significantly attenuated by LY294002. These findings suggest that ATF4 plays a crucial role in endothelial cell-mediated neovascularization under post-MI hypoxia by modulating the PI3K/AKT signaling pathway.

Recent studies have highlighted the role of ferroptosis in neuronal damage during neonatal hypoxic-ischemic encephalopathy (HIE). Nuclear protein 1 (NUPR1), a newly identified crucial modulator of ferroptosis, remains unexplored in the context of HIE. This study aimed to investigate whether NUPR1 modulates ferroptosis and influences hypoxic-ischemic brain injury through specific molecular mechanisms. NUPR1-knockdown neurons presented increased sensitivity to Erastin-induced neuronal ferroptosis, whereas NUPR1 overexpression conferred resistance. Notably, silencing NUPR1 exacerbated OGD/R-induced neuronal damage and ferroptosis, as evidenced by increased lipid peroxidation, malondialdehyde (MDA) levels, and iron concentrations, as well as decreased glutathione (GSH) levels and altered expression of ferroptosis-related proteins (elevated PTGS2 and reduced GPX4). Conversely, NUPR1 overexpression alleviated OGD/R-induced neuronal damage and ferroptosis. HIE animal model experiments demonstrated that NUPR1 overexpression mitigated brain damage, reduced infarct size, and decreased brain edema, which were correlated with diminished ferroptosis markers. Furthermore, NUPR1 knockdown reduced ferritin heavy chain 1 (FTH1) expression, whereas NUPR1 overexpression increased FTH1 levels, indicating a regulatory role in iron metabolism. Silencing FTH1 reversed the inhibitory effect of NUPR1 on neuronal ferroptosis. Collectively, our findings indicate that NUPR1 protects against ferroptosis in HIE, making it a potential therapeutic target for reducing neuronal injury associated with this condition. NUPR1 suppresses neuronal ferroptosis by increasing FTH1 expression and improving iron storage, enhancing our understanding of the mechanisms involved in ferroptosis in neonatal HIE.