Cell Biochemistry and Biophysics最新文献

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.

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.

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.

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.

Hepatic steatosis, commonly referred to as fatty liver disease, is defined by the abnormal buildup of fat within liver cells. Currently, primary treatments mainly focus on lifestyle changes, underscoring a lack of direct pharmacological options. Passion fruit seed extract (PFSE) has been reported to decrease hepatosteatosis; however, the mechanism underlying this effect has not been clarified. Therefore, the objective of this research was to investigate the effects and mechanisms of action of PFSE against oleic acid (OA)-induced hepatosteatosis in HepG2 cells. OA-induced HepG2 cells were analyzed by using various cell-based experiments, including assessments of cytotoxicity, reactive oxygen species (ROS) production, apoptosis, and protein and gene expression. LC-MS-MS analysis showed that PFSE contains a variety of phytochemical compounds such as alkaloids, flavonoids, stilbenoids, coumarins, terpenoids, lipids, and fatty acid derivatives, which have the potential to exhibit various pharmacological activities. In this study, PFSE demonstrated antioxidant, anti-inflammatory, and lipid metabolism-regulating activities. It also influenced key genes related to lipid metabolism, including SREBP-1c, ACC, FASN, PPARα, CPT-1A, LPL, SCD1, and LDLR. The positive effects of PFSE on OA-induced hepatic steatosis in HepG2 cells were modulated through the Akt and ERK signaling pathways, suggesting that PFSE may offer a comprehensive approach to managing hepatic steatosis.

In the regulation of gene expression, epigenetic factors, including non-coding RNAs (ncRNAs) play a role in genetics. Among the ncRNA family, microRNAs (miRNAs) have gained significant attention for their involvement in post-transcriptional gene regulation, with profound implications for both normal and pathological processes including neurological diseases such as hypoxic-ischemic brain injury. A specific miRNA, called miR-122-5p, has gained attention in hypoxic-ischemic conditions, where it modulates critical pathways such as inflammation, oxidative stress, and neuronal survival. The purpose of this review is to highlight recent advances in the biogenesis, expression, and regulation of miR-122-5p, focusing on its role in hypoxic-ischemic conditions and its potential as a therapeutic target. We first studied the therapeutic strategies and potential clinical applications of miR-122-5p, our research showing it interacts with key transcription factors, such as HIF-1α and NF-κB, influencing cellular responses to low oxygen levels. Our findings revealed that miR-122-5p plays a vital role in hypoxic-ischemic brain injury, with its abnormal levels strongly associated with increased brain damage and neuroinflammation, suggesting its potential as a promising therapeutic target. Furthermore, miR-122-5p influences various biological processes in the brain, such as metabolism and blood vessel formation. The use of miR-122-5p inhibitor has been shown to increase autophagy, reduce apoptosis, and decrease oxidative stress and inflammation, thereby protecting neurons and improving outcomes in hypoxic encephalopathy by targeting multiple genes related to these processes. Conversely, miR-122-5p mimics exacerbate oxidative stress and reduce autophagy. These findings highlight the therapeutic potential of miR-122-5p inhibition in reducing brain injury and promoting recovery in hypoxic-ischemic encephalopathy through enhanced neuroprotective mechanisms and the suppression of harmful cellular processes. However, further experimental studies are needed to fully understand the therapeutic potential of targeting miR-122-5p and its related genes in hypoxic-ischemic encephalopathy.