Cell Biochemistry and Biophysics最新文献

This study aims to elucidate the role of miR-378a-3p in facilitating the proliferation and differentiation of synovium-derived mesenchymal stem cells (SMSCs) into chondrocytes. The effects of overexpressing miR-378a-3p on SMSCs were investigated through histological analysis, quantitative PCR, and western blotting. Then we identified binding sites of miR-378a-3p with BMP2 through Gene Ontology and Kyoto Encyclopedia of Genes and Genomes analyses and predictions from the RegRNA 2.0 database. Subsequently, BMP2 was confirmed as the target by which miR-378a-3p promotes the chondrogenic differentiation of SMSCs using a luciferase reporter gene assay and an miR-378a-3p RNA interference plasmid. Finally, by constructing a rat model with articular cartilage damage, we detected the reparative effects of miR-378a-3p overexpression on cartilage damage. Additionally, we verified the mechanism by which miR-378a-3p promotes chondrogenic differentiation in SMSCs. MiR-378a-3p enhances the proliferation and differentiation of SMSCs into chondrocytes by modulating the BMP2-Smad signaling pathway, thereby facilitating repair processes for articular cartilage injuries in rats. Notably, knockdown of BMP2 diminished the reparative efficacy of miR-378a-3p on articular cartilage damage. Upregulation of miR-378a-3p promotes chondrogenic differentiation in SMSCs through activation of the BMP2-Smad pathway, positioning it as a potential therapeutic target for osteoarthritis.

This study aimed to investigate the role and underlying mechanisms of the platelet-derived growth factor (PDGF)/protein kinase B (AKT) signaling pathway in pressure overload-induced ventricular remodeling. Ventricular remodeling, a critical pathological process in heart failure, is commonly triggered by pressure overload. While PDGF is known to promote cell proliferation and growth, the AKT pathway is crucial for cell growth, survival, and metabolism. However, the specific role of the PDGF/AKT pathway in pressure overload-induced ventricular remodeling remains unclear. Thus, this study aimed to elucidate the precise mechanisms of PDGF/AKT involvement in this process using animal models and cell experiments. 45 female C57BL/6 mice were utilized, randomly divided into three groups: model group (M group, n = 15), control group (C group, n = 15), and experimental group (E group, n = 15). M group mice underwent thoracotomy without aortic constriction (AC). C group mice received phosphate-buffered saline (PBS) and dimethyl sulfoxide (DMSO) treatment following AC surgery. E group mice were treated with the PDGF receptor inhibitor AG1296 and PBS solution after AC surgery. Additionally, 293 T cells were categorized into three groups: PDGF shRNA transfected group (downregulating PDGF expression, D group), PDGF overexpression group (B group), and control group (NV group). Left ventricular end-systolic volume (LVESV) and ejection fraction (FS) of the mice were measured via echocardiography. Western blot analysis was conducted to assess the expression levels of p-AKT and t-AKT in myocardial tissues. Furthermore, myocardial cell area was measured using hematoxylin and eosin (HE) staining and image analysis software. The LVESV in the C group was significantly higher than in the M and E groups (48.32 ± 3.08 mL vs. 18.24 ± 3.19 mL and 25.44 ± 3.12 mL, P < 0.05). The FS in the C group was significantly lower compared to the M and E groups (21.18 ± 2.99% vs. 42.45 ± 3.02% and 26.89 ± 2.54%, P < 0.05). Western blot analysis revealed that p-AKT and t-AKT levels were significantly elevated in the C group and PDGF overexpression group (B group) compared to the M and PDGF shRNA groups (D group) (P < 0.05). HE staining showed a significant increase in myocardial cell cross-sectional area in the C and D groups, with the most pronounced enlargement in the D group (P < 0.05). PDGF facilitates pressure overload-induced ventricular remodeling and myocardial fibrosis. Inhibition of the PDGF/AKT signaling pathway effectively mitigates myocardial cell hypertrophy and ventricular remodeling. These findings offer novel potential targets and therapeutic strategies for the treatment of pressure overload-related heart failure.

Type 2 diabetes mellitus (T2DM) is usually depicted by relative insulin deficiency, raised blood glucose levels, and the predominant risk factor, insulin resistance. Hence, the development of insulin sensitizer drugs targeting PPAR-γ receptors has expanded enormous interest as an attractive choice for T2DM treatment. Thiazolidinediones (TZD) enhance insulin sensitivity either by directly functioning on gene transcription of the PPARγ receptor related to glucose homeostasis or by systemic sensitization of insulin and, therefore, improved hyperglycemia in a wide range of patients. However, severe complications and adverse effects of TZDs necessitate the development of an efficacious and reliable insulin sensitizer from alternative resources. On the contrary, Nature is a rich source of anticipated effective and safer medicine; more than fifty percent of drugs on the market are developed from natural products. Hence, searching for a new PPAR-γ agonist from bioactive secondary compounds of medicinal plants along with greater efficacy and safety is a recognized and consistent tactic for developing novel antidiabetic agents. Pulicaria jaubertii is a fragrant perennial aromatic plant with anti-inflammatory, antidiabetic, antimicrobial, antimalarial, and insecticidal properties. The current study was designed to use a computer-aided drug design to explore the best antidiabetic compounds from P. jaubertii. Herein, the molecular docking study of 80 investigated ligands against the PPAR-γ receptor identifies the highest docking score for five ligands ranging from -8.9 kcal/mol to 8.0 kcal/mol, which is also more significant than the standard drug pioglitazone (-7.7 kcal/mol) determined by the PyRx 8.0 virtual screening software. GLN286, CYS285, SER289, TYR473, MET364, ARG288, ILE341, and LEU333 residues are found to be significant contributors to the non-bonded interaction between ligands and receptors. Molecular electrostatic potential (MEP), DFT, molecular orbital (MO), ADMET, and toxicological analyses were performed on the selected five high-scored ligands of P. jaubertii. Results documented that all investigated ligands, especially L4, show considerably excellent profiles in molecular docking, MEP, DFT, MO, ADMET, and toxicological predictions, suggesting our drug-designing approaches may contribute to the development of a novel antidiabetic drug for the treatment of T2DM from natural resources.

Neurodegenerative disorders are associated with the accumulation of disease-related proteins intracellularly and extracellularly. Extracellular chaperones play a crucial role in clearing the extracellularly accumulated proteins. In this study, we observed the extracellular chaperone-like potential of BSA at physiological concentrations on model protein cytochrome c (cyt c). Kinetics of heat-induced aggregation of cyt c suggest the nucleation independent first order aggregation kinetics. Aggregation of cyt c was studied in the presence of varying concentrations of BSA to assess its chaperone nature. At lower concentrations of BSA when the sub molar ratio of cyt c:BSA are 1:0.6 and 1:1.2, heat-induced unfolded cyt c promotes the aggregation of BSA. However, as the ratio of cyt c:BSA increases to 1:1.8, the aggregation of cyt c is reduced. When the concentration of BSA reaches physiological levels, yielding a cyt c:BSA ratio of 1:2.4, the rate of aggregation drastically decreases reflecting its chaperone potential. These observations indicate that under physiological conditions, macromolecular crowding stabilizes the native structure of both proteins and enhances their interaction that results in the reduced aggregation of cyt c. Additionally, the presence of the phytochemical chlorogenic acid at a sub-molar ratio of 1:1 stabilizes cyt c and prevents its unfolding and facilitates the binding of cyt c to BSA at physiological concentrations. This interaction further decreases the overall aggregation of cyt c and stabilizes its native fold.

Growth regulatory factors (GRFs) are transcription factors that encode the proteins involved in plant growth and development. However, no comprehensive analysis of Vitis vinifera GRF genes has yet been conducted. In the current study, we performed a genome-wide analysis of the GRF gene family to explore the VvGRF gene's role in Vitis vinifera. We identified 30 VvGRF genes in the Vitis vinifera genome, localized over 20 chromosomes. Based on evolutionary analysis, 49 GRF genes (nine AtGRF, ten FvGRF, and 30 VvGRF) were clustered into six groups. Many cis-elements involved in light control, defense, and plant growth have been identified in the promoter region of VvGRF genes, and multiple miRNAs have been predicted to be involved in regulating VvGRF gene expression. Protein-protein interaction analysis showed that nine VvGRF proteins formed a complex protein interaction network. Furthermore, the gene expression analysis of VvGRF revealed that VvGRF-5 and VvGRF-6 were highly upregulated suggesting that these genes are involved in biotic responses. This study provides comprehensive insights into the functional characteristics and occurrence of the VvGRF gene family in Vitis vinifera, which may be applied in breeding programs to enhance the growth of Vitis vinifera varieties under stress and growth changes.

Major depressive disorder (MDD) is a severe mental disorder with largely unknown mechanisms. Carbonic anhydrases convert CO₂ to carbonates and protons, playing roles in various brain functions. Carbonic anhydrase 1 (Car1) is particularly abundant and may be linked to microbiota at interstitial sites. We developed Car1-deficient mice to explore the relationship between depression-like behaviors and gut microbiota. Behavioral tests confirmed depression-like behavior in Car1^-/- mice. Fecal samples from Car1^-/- and WT mice were collected, and 16S rRNA gene sequencing identified distinct microbiota components between the groups. Car1^-/- mice exhibited significantly increased immobility in the tail suspension test (TST) compared to WT mice. The gut microbiota composition differed at the phylum level in p_Bacteroidetes, p_Verrucomicrobia, p_Firmicutes, and p_Tenericutes. At the family level, Car1^-/- mice had significantly different abundances in eight microbiota groups compared to WT mice. Car1 deficiency is associated with depressive-like behavior and gut microbiota dysbiosis, potentially linked to depressive-like phenotypes.

Periodontitis is a prevalent condition characterized by inflammation and tissue destruction within the periodontium, with hypoxia emerging as a contributing factor to its pathogenesis. Hypoxia-inducible factor 1α (HIF-1α) has a crucial role in orchestrating adaptive responses to hypoxic microenvironments and has been implicated in various inflammatory-related diseases. Understanding the interplay between HIF-1α, matrix metalloproteinases (MMPs), and inflammatory responses in periodontitis could provide insights into its molecular mechanisms. We investigated the relationship between HIF-1α, MMP2, and MMP9 in gingival crevicular fluid (GCF) and periodontal ligament stem cells (PDLSCs) from periodontitis patients. The expression levels of HIF-1α, MMP2, MMP9, and inflammatory factors (IL-6, IL-1β, TNF-α) were assessed using enzyme-linked immunosorbent assay (ELISA) and real-time PCR (RT-PCR). Additionally, osteogenic differentiation of PDLSCs was identified by alkaline phosphatase activity. Significantly elevated levels of HIF-1α, MMP2, and MMP9 were observed in GCF of periodontitis patients compared to controls. Positive correlations were found between HIF-1α and MMP2/MMP9, as well as with IL-6, IL-1β, and TNF-α. Modulation of HIF-1α expression in PDLSCs revealed its involvement in MMP2/9 secretion and inflammatory responses, with inhibition of HIF-1α mitigating these effects. Furthermore, HIF-1α inhibition alleviated the reduction in osteogenic differentiation induced by inflammatory stimuli. Our findings elucidate the regulatory role of HIF-1α in MMP expression, inflammatory responses, and osteogenic differentiation in periodontitis. In conclusion, targeting HIF-1α signaling pathways may offer therapeutic opportunities for managing periodontitis and promoting periodontal tissue regeneration.

In male rats, the flaxseed oil (FS-oil) modulatory properties were investigated on diazinon (DZN)-induced nephrotoxicity. Adult male Wistar rats were divided randomly into five groups. To induce nephrotoxicity, animals received DZN (70 mg/kg/day, p.o.). Also, treatment groups received FS-oil (100 and 200 mg/kg/day, p.o.). The animal treatment was 28 consecutive days. On the 29th day, serum and kidney tissue samples were removed and serum levels of the creatinine, blood urea nitrogen (BUN), malondialdehyde (MDA), glutathione peroxidase (GPx), and catalase (CAT), were measured. Also, hematoxylin and eosin (H&E) staining was applied for histological studies. DZN significantly increased the BUN, creatinine, and MDA levels compared to the control group. Besides, DZN significantly decreased the GPx and CAT activity in the kidney tissue. However, the modulatory effects of FS-oil were observed by improving renal enzyme factors, inhibiting oxidative stress, and histological change. This study demonstrated that FS-oil ameliorated DZN-induced nephrotoxicity and can be used as a preventive agent against DZN toxicity because of the FS-oil antioxidant characteristics.

Parkinson's disease (PD) is a prevalent neurodegenerative disorder for which novel treatment approaches are continuously sought. This study investigates the role of high-mobility group A1 (HMGA1) in modulating inflammatory responses and oxidative stress injury in PD. We utilized the murine dopaminergic neuronal cell line MN9D, treating cells with 1-methyl-4-phenylpyridinium ion (MPP⁺) to mimic PD conditions. The expression levels of HMGA1 and insulin receptor substrate 2 (IRS2) were measured using quantitative polymerase chain reaction and Western blot assay. Cell damage was assessed with cell counting kit-8 and lactate dehydrogenase assays. Inflammatory response and oxidative stress were evaluated by quantifying interleukin (IL)-1β, IL-6, tumor necrosis factor-α, reactive oxygen species, superoxide dismutase, and malondialdehyde (MDA) levels using enzyme-linked immunosorbent assay and commercial kits. The binding interaction between HMGA1 and IRS2 was analyzed using chromatin immunoprecipitation (ChIP) and dual-luciferase reporter assays. Our findings revealed that MPP⁺ treatment increased the expression of HMGA1 and IRS2. Downregulation of HMGA1 enhanced cell viability, reduced inflammation, and mitigated oxidative stress in MPP⁺-induced cells. Further investigation demonstrated that HMGA1 bounded to the IRS2 promoter, enhancing IRS2 expression. Overexpression of IRS2 counteracted the protective effects of HMGA1 downregulation. In conclusion, HMGA1 exacerbates MPP⁺-induced cell damage by activating IRS2 transcription, which in turn heightens inflammation and oxidative stress. These findings suggest that targeting HMGA1 could be a potential therapeutic strategy for PD.

Silene vulgaris (Moench) Garcke and Stellaria media (L.) Vill is a perennial wild weed species belonging to the Caryophyllaceae family and is widely available and abundant in the environment. The present study has aimed to evaluate the anti-inflammatory potential of two underutilized wild edible plants, Silene vulgaris (Moench) Garcke and Stellaria media (L.) Vill. fractions employing in-vitro COX inhibitory assay. Invitro COX-2 inhibitory potential of MESV and MESM fractions was carried out using BioVision^R "COX Activity Assay Kit (Fluorometric)". LC-MS analysis of selected fractions was conducted to identify bioactive compounds that were further validated for their affinity determination toward target enzymes employing molecular docking studies using the LibDock program. In-vitro COX inhibitory assay revealed that hexane fraction of S. vulgaris (HFSV) and hexane fraction of S. media (HFSM) caused impressive inhibition of COX-2 enzyme with IC₅₀ values 1.38 µg/mL and 1.51 µg/mL respectively. Further, LC-MS analysis revealed the presence of 46 compounds in HFSV and 44 compounds in HFSM respectively. Amongst identified bioactive compounds in HFSV and HFSM, sinapinic acid and syringic acid showed good docking scores with COX-2 i.e., 89.256, and 82.168 respectively. Also, the availability of chrysin in HFSM and rhamnetin in HFSV exhibited good docking scores i.e., 115.092, and 112.341 with a selective affinity towards COX-2. The findings of in-vitro COX Inhibitory Activity and molecular docking studies highlighted the impressive anti-inflammatory properties of S. vulgaris and S. media, and require further investigations to establish them as therapeutic candidates in the management of inflammation and related issues.