Cerebral ischemia/reperfusion (I/R) injury is a major cause of neurological disability. This study investigated the role of lncRNA PVT1 in I/R-induced cerebral injury and its clinical relevance. MCAO/reperfusion mouse and OGD/R neuronal cell models were established. Neurological function, infarct volume, brain edema, and neuronal apoptosis were assessed. For clinical validation, 50 acute ischemic stroke patients and 50 healthy controls were enrolled. Serum PVT1/miR-187-3p were measured by qRT-PCR; plasma MyD88 and inflammatory cytokines (TNF-α, IL-6, IL-18) by ELISA. Machine learning (Random Forest) and correlation analyses were performed to evaluate diagnostic utility and relationships with clinical severity (NIHSS score, infarct volume). PVT1 was significantly upregulated under I/R both in vivo and in vitro. Clinically, AIS patients showed markedly elevated PVT1 (3.92 vs. 1.02, p < 0.0001) and reduced miR-187-3p (0.42 vs. 1.01, p < 0.0001), with increased MyD88 and inflammatory cytokines (all p < 0.001). The Random Forest model achieved AUC = 0.92, with PVT1 as the top predictor. PVT1 correlated positively with MyD88 (r = 0.68, p < 0.001) and NIHSS score (r = 0.61, p < 0.001). PVT1 silencing in MCAO mice significantly improved neurological function, reduced infarct volume and brain edema, and attenuated neuronal apoptosis (all p < 0.01). Mechanistically, PVT1 sponged miR-187-3p to derepress MyD88/TLR4 signaling, which was reversed by miR-187-3p restoration. In vivo rescue experiments confirmed that miR-187-3p antagomir reversed the protective effects of PVT1 silencing across multiple endpoints. PVT1 inhibition protects against cerebral I/R injury by restoring miR-187-3p and blocking the MyD88/TLR4 pathway. PVT1 and its downstream axis are associated with stroke diagnosis and severity, and represent promising therapeutic targets and diagnostic biomarkers for ischemic stroke.
This study presents a comprehensive evaluation of the micropropagation of ‘Duke 7’ and ‘Toro Canyon’ avocado (Persea americana Mill.) clonal rootstocks. The effects of different basal media (WPM, MS, B5, and DKW) and the number of subcultures on the morphological development and physiological disorder parameters of nodal segments were evaluated. The best overall shoot development was observed on WPM for ‘Duke 7’ and on MS medium for ‘Toro Canyon’. Subsequently, the effects of two different cytokinins (BAP and mT) and their concentrations (1.0 and 2.0 mg L⁻1) on the same parameters were evaluated over three successive subcultures in the selected media. The highest shoot proliferation in both rootstocks was obtained with 2.0 mg L⁻1 BAP. However, the best overall shoot development was observed with 1.0 mg L⁻1 BAP for ‘Duke 7’ and 1.0 mg L⁻1 mT for ‘Toro Canyon’. Increasing the number of subcultures promoted shoot proliferation but also increased physiological disorders. The effects of two auxins (IBA and NAA) at different concentrations (0.0, 0.5, 1.0, 2.0, and 3.0 mg L⁻1) on in vitro rooting were evaluated. The highest rooting percentages were 45.00
Tendon stem/progenitor cells (TSPCs) play essential roles in tendon homeostasis and repair, but their regenerative capacity is compromised by oxidative stress-induced senescence and the associated loss of stemness. Here, we identified PEP472, a previously uncharacterized micropeptide encoded by a small open reading frame (smORF) within the long non-coding RNA (lncRNA) MSTRG.44472.1, as a regulator of TSPC function under oxidative stress. The expression of MSTRG.44472.1 decreased in an H₂O₂-induced TSPC senescence model, and the coding potential of its smORF and translation into PEP472 were subsequently validated. Functional analyses demonstrated that PEP472 attenuated H₂O₂-induced senescence, preserved TSPC stemness, and promoted tenogenic differentiation. Mechanistically, PEP472 increased SIRT1 protein abundance by suppressing proteasome-dependent SIRT1 degradation and enhancing SIRT1 stability. Consistent with a role for SIRT1 in mediating the protective effects of PEP472, pharmacological inhibition of SIRT1 with EX-527 markedly attenuated the anti-senescence effects of PEP472. Together, these findings identify a previously unrecognized MSTRG.44472.1-PEP472-SIRT1 regulatory axis in TSPCs and demonstrate that PEP472 attenuates oxidative stress-induced senescence and preserves TSPC stemness through SIRT1 stabilization. This study highlights PEP472 as a promising micropeptide-based biologic candidate for preventing senescence-associated TSPC dysfunction and supporting tendon regenerative repair.
Garcinia mangostana L. is a tree species in the Clusiaceae family, commonly known as mangosteen. While its fruit is consumed worldwide, it has a long history of use as a medicinal plant, particularly in Southeast Asia. It exhibits potent anti-inflammatory properties and has been used as a treatment for various symptoms such as acne and diarrhea. Furthermore, G. mangostana is characterized by the production of a group of highly modified tricyclic polyphenols known as xanthones. Many of these possess multiple hydroxyl and prenyl groups, and exhibit a wide range of pharmacological activities. Although the discovery of new compounds from G. mangostana continues to be actively reported, there are only a few comprehensive reviews summarizing its metabolic compounds and their pharmacological activities. Furthermore, in vitro cultures are being conducted as a strategy to obtain these unique secondary metabolites. In recent years, callus induction from G. mangostana and the profiling of some metabolites have been reported. However, to the best of current knowledge, no review on in vitro tissue culture of G. mangostana has yet been published. Therefore, the objective of this review is to comprehensively organize the diverse xanthones produced by G. mangostana, along with their pharmacological activities and the progress in strategies for obtaining G. mangostana-derived metabolites through in vitro tissue culture. This review will provide valuable insights for the discovery of novel lead compounds for drug discovery and further advances in cell culture.
Objective. This study aimed to investigate the protective effect of Calycosin, a major bioactive flavonoid derived from Astragalus membranaceus, on TGF-β1-induced pulmonary fibrosis in human embryonic lung fibroblast (MRC-5) cells and to elucidate the underlying mechanisms. Methods. Cell viability was assessed using the MTT assay to determine non-cytotoxic concentrations of CA for subsequent experiments. TGF-β1-stimulated MRC-5 cells were treated with varying doses of CA. The anti-fibrotic effects were evaluated by measuring the expression of fibrosis markers (α-SMA and Collagen I) via Western blotting and immunofluorescence. Intracellular reactive oxygen species (ROS) levels and the activities of antioxidant enzymes were measured to assess oxidative stress. Cell migration was monitored using a scratch wound assay. The involvement of the TLR4/MyD88/NF-κB signaling pathway and the expression of downstream inflammatory cytokines (IL-17A, IL-6, TNF-α) were analyzed by Western blot and quantitative real-time PCR (qRT-PCR). The TGF-β receptor inhibitor SB-431542 was used as a comparative control. Results. Calycosin treatment significantly attenuated TGF-β1-induced activation of MRC-5 cells, as evidenced by the marked downregulation of α-SMA and Collagen I expression. CA also ameliorated oxidative stress by reducing ROS generation and restoring glutathione (GSH) levels and total superoxide dismutase (T-SOD) activity. Furthermore, CA inhibited the enhanced migratory capacity of fibroblasts induced by TGF-β1. Mechanistically, CA downregulated the protein expression of key components (TLR4, MyD88, phospho-NF-κB p65, phospho-IκBα) in the TLR4/MyD88/NF-κB pathway and reduced the mRNA and protein levels of the pro-fibrotic inflammatory cytokines IL-17A, IL-6, and TNF-α. The efficacy of CA was comparable to that of SB-431542. Conclusion. Calycosin effectively attenuates TGF-β1-induced fibroblast activation, oxidative stress, and ECM remodeling in pulmonary fibroblasts. Its anti-fibrotic effects are primarily mediated through the suppression of the TLR4/MyD88/NF-κB signaling pathway and the reduction of the pro-inflammatory microenvironment. These findings suggest that Calycosin holds promising potential as a therapeutic agent for silicosis and other pulmonary fibrotic diseases.
