Journal of Physiology and Pharmacology最新文献

Human cell line HS-27A represents an immortalized subpopulation of human stromal cells derived from bone marrow. HS-27A cells meet the criteria set by the International Society of Cell Therapy (ISCT) for the classification as mesenchymal stem cells (MSCs) by expression of surface molecules CD73, CD90, CD105 and HLA-ABC with no expression of CD14, CD31, CD34, CD45 and HLA-DR. We hypothesized that these cells may undergo osteogenesis similar to human bone marrow-derived stromal cells (BMSCs) and serve as a model of osteogenic cell responses under normal and inflammatory conditions. HS-27A cells were treated with: standard osteogenic factors, i.e., ascorbic acid (Asc), dexamethasone (Dex) and β-glycerophosphate (BGP); recombinant human bone morphogenetic protein 2 (rhBMP-2), MEK1/2 kinase inhibitor (PD98059) and/or lipopolysaccharide (LPS). Cells were examined for alkaline phosphate (ALP) activity, mRNA expression of osteoblastic markers (qRT-PCR) and mineralization of extracellular matrix. Besides, we analyzed mRNA expression of proinflammatory cytokines IL-1β and TNF-α in LPS-treated cells and evaluated transfection efficiency of these cells with Lipofectamine 3000 for potential future genetic manipulations. We determined that HS-27A cell line increase alkaline phosphatase activity (P<0.05), while mineralization of extracellular matrix remains low. Cells treatment with rhBMP-2 and PD98059 resulted in increased mRNA levels for osteogenesis-related transcription factor MSX-2, bone sialoprotein and osteocalcin (all P<0.05). In osteogenic cultures with and without rhBMP-2, addition of LPS led to increased ALP activity, mRNA levels for collagen type I and osteocalcin as well for IL-1β and TNF-α (all P<0.05). We also show that HS-27A cells can be transfected with phMGFP plasmid using Lipofectamine 3000 with low efficiency that may be sufficient for some genetic manipulations. Thus, HS-27A cell line appear as a useful in vitro cell culture model to study short-term osteogenic and inflammatory-related responses of human bone marrow stromal cells.

The signal transducer and activator of transcription-3 (STAT-3) is a perilous transcription factor that regulates various proliferation and anti-apoptosis factors in prostate cancer cells. Therefore, inhibiting STAT-3 signaling is considered a potential therapeutic approach for treating prostate cancer. This study investigates the effects of borneol (BNL) on the proliferation and apoptosis of human prostate cancer cells by blocking Janus kinase (JAK) and STAT-3 expression. Human prostate cancer (PC-3) cells were treated with varying concentrations of BNL (10, 20, 30 μM) for 24 hours. Subsequent analyses included MTT based cytotoxicity, reactive oxygen species (ROS) production measured by 2,7'-dichlorodihydrofluorescein diacetate (DCFH-DA) staining, and apoptosis morphological examination was studied by acridine orange/ethidium bromide (AO/EtBR) staining. BNL treatment mediated protein expression of proliferation and apoptosis-related proteins associated with the JAK-STAT-3 pathways was investigated by Western blot. Results of this study indicates that BNL treatment with PC-3 cells induces cytotoxicity, increases ROS production, and causes apoptotic morphological changes in a concentration-dependent manner. BNL significantly reduced the expression of cell proliferation markers such as cyclin-D1, cyclin-D2 and cyclin-E1 (P<0.05) compared to untreated PC-3 control cells. BNL treatment enhanced apoptosis rates by observed overexpression of Bcl-2-associated X protein (Bax), caspase-3 and down regulation B-cell leukemia/lymphoma 2 (Bcl-2) (P<0.05) expression in PC-3 cells. Additionally, BNL reduced interleukin-6, JAK1, and STAT3 phosphorylation ((P<0.05) in PC-3 cells that obstructing the expression of proliferation and anti-apoptotic proteins in PC-3 cells. Thus, BNL may be a therapeutic agent against prostate cancer by blocking the STAT3 signaling axis.

Coronavirus disease 2019 (COVID-19) has been associated with various systemic complications, including potential impacts on ocular health. Recent studies have suggested that COVID-19 may lead to changes in retinal structure, particularly in the neuroretinal and retinal nerve fiber layers (RNFL). This study aimed to examine changes in neuroretinal and RNFL of the optic dics based on optical coherence tomography (OCT) in patients hospitalized due to COVID-19 bilateral pneumonia at 2 time points after discharge. A prospective study involved 49 patients with COVID-19 bilateral pneumonia hospitalized between March and May 2021. Baseline ocular evaluations were conducted 2 months post-discharge, with follow-up examinations 6 months later. Retinal parameters, including RNFL and ganglion cell layers (GCL), were assessed. Control group of healthy individuals also underwent similar ophthalmic examinations for comparative analysis. We found that the average thickness of the retinal nerve fiber layer (RNFL) of the optic disc was lower in the COVID-19 group than in controls (p≤0.01). The ganglion cell layer (GCL) was thicker in the inner inferior ring (p=0.008) but thinner in the outer superior, outer nasal, and outer inferior rings (p=0.044, p<0.01, and p<0.01, respectively). OCT parameters were assessed according to sex. At 6 months, the RNFL of the inner inferior and outer temporal rings was thinner in women than in men (p=0.022 and p=0.020, respectively). The GCL of the inner temporal and outer temporal rings was also thinner in women than in men (p=0.004 and p=0.005, respectively). In conclusion: at 6-month follow-up, RNFL and GCL thickness was significantly lower in some areas of the retina compared with baseline. COVID-19 seems to cause changes in the macular retina, highlighting the need for ophthalmologic screening of patients with a history of SARS-CoV-2 infection.

The endocannabinoid system (ECS) and nociceptin receptor (NOP) have been implicated in the pathology of inflammatory bowel diseases (IBD) mediating pain and alleviating inflammation. In this study we searched for the possible activation of ECS and NOP system and the correlation between CB1, CB2 and NOP receptors in IBD patients. Patients diagnosed with IBDs who underwent colonic surgical resection or biopsy at colonoscopy and control group (patients without diagnosis of IBD, which colonoscopy for the different medical indications) were recruited into the study. In surgical specimen the quantification of endocannabinoids was obtained by liquid chromatography-atmospheric pressure chemical ionization-mass spectrometry. In biopsy specimen, the expression of genes encoding CB1, CB2, and NOP receptors was determined with real-time RT-PCR. The relative expression of CB1 and CB2 receptors in human samples with IBD compared to the unrelated controls (HC) group was reduced compared to the controls, but no differences in relative expression of NOP receptor were detected. Statistical significance difference was reached only between the level of a relative expression of CB1 receptor in ulcerative colitis (UC) and HC groups (498 (57.45-1890) and 4946 (2098--12818) (P<0.05)). A statistically significant positive correlation between the relative expression of CB1 and NOP receptors (r=0.83, P<0.05) as well as CB2 and NOP receptors in Crohn's disease (CD) (r=0.87, P<0.05) was found. Several endocannabinoids were significantly (P<0.05) increased in tissue collected from UC and CD patients in comparison to controls. CB1 and CB2 but not NOP receptors were found to be downregulated in IBD. Correlation of CB1, CB2 and NOP expression may suggest their common roles and shared molecular pathways in CD. Upregulated level of several endocannabinoids may point out to their role in IBD. Their estimation may be possibly useful in IBD diagnostics.

Baicalin, a predominant bioactive flavonoid derived from the traditional Chinese medicinal herb Scutellaria baicalensis Georgi, has garnered significant attention. Ferroptosis, a relatively novel form of programmed cell death, implicates critical signaling pathways, notably those involving nuclear factor E2-related factor 2 (Nrf2) and glutathione peroxidase 4 (Gpx4). Consequently, this study aims is to elucidate whether baicalin mitigates intestinal tissue damage by modulating the Nrf2-Gpx4 signaling pathway in the context of intestinal ischemia-reperfusion (II/R) injury, thereby influencing iron deposition. For this purpose it was established an II/R rat model and a cellular hypoxia-glucose deficiency/reoxygenation (OGD/R) model and administered BA to the II/R rats and OGD/R model cells. It was utilized HE staining and probe staining techniques to assess intestinal injury and iron overload, respectively and employed RT-qPCR to measure the mRNA expression levels of inflammation-related genes (interleukin-1β, interferon-γ, interleukin-4, and interleukin-10) and iron deficiency-related genes (Nrf2, Gpx4, and xCT). The cell counting kit-8 (CCK-8) assay is employed to assess cell viability, while fluorescent probes are utilized to evaluate mitochondrial membrane potential. Colorimetric methods are applied to quantify intracellular oxidative stress-related indicators (reactive oxygen species (ROS) and malondialdehyde (MDA)) levels. Flow cytometry is used to determine cell membrane lipid ROS levels. Immunofluorescence techniques are implemented to examine intestinal tight junction proteins, such as ZO-1 and Occludin. Additionally, protein immunoblotting is conducted to measure markers of iron deposition (Gpx4 and xCT). In in vivo studies, BA treatment mitigated intestinal damage in II/R rats, inhibited intestinal iron ion overload, and elevated inflammatory levels and oxidative stress. Furthermore, BA treatment reduced the loss of tight junction proteins ZO-1 and Occludin expression levels induced by II/R. In vitro studies demonstrated that BA significantly attenuated OGD/R-induced iron overload, decreased cell viability, and mitigated mitochondrial membrane potential loss. Mechanistically, BA exerts its protective effect against iron overload-induced cellular damage by activating the Nrf2-Gpx4 signaling pathway. However, these effect was significantly counteracted by the use of Nrf2-GPX4 pathway inhibitors. The positive regulation of Nrf2-Gpx4 by BA can reduce ferroptosis and alleviate II/R in rats.

Asthma is a prevalent chronic inflammatory airway disease that affects both adults and children. Inflammation-induced airway remodeling can lead to irreversible damage to the airways. Traditional Chinese medicine (TCM) plays a significant role in healthcare, offering potential improvements for chronic airway inflammation associated with asthma. The objective of this study is to investigate the efficacy of Pingchuan formula (PCF) in treating asthma and explore its underlying mechanisms. The asthmatic model mice were sensitized on days 1 and 7, followed by aerosolized OVA challenge for a total of 21 times starting from day 14, spanning weeks 3 to 9. PCF was administered daily after the first challenge. The mice were orally dosed daily based on their individual body weights for a continuous period of 47 days. The examined items encompassed proteomic analysis of the target proteins impacted by PCF. The levels of interleukins IL-1β, IL-18, and transforming growth factor-β (TGF-β) in bronchoalveolar lavage fluid (BALF) were measured using ELISA. Immunohistochemistry was employed to assess the expression levels of nucleotide-binding domain, leucine-rich-containing family, pyrin domain-containing-3 (NLRP3), caspase-1, and TGF-β in the airways. Western blotting and real-time quantitative PCR analysis were conducted to determine NLRP3 and caspase-1 expression levels. Additionally, Western blotting was performed to evaluate C-Jun N-terminal kinase (JNK), phosphorylated JNK (p-JNK), matrix metalloproteinase-9 (MMP-9), N-cadherin, E-cadherin, and tenascin C (T-NC) expression. Compared to the asthma model group, histological analysis using hematoxylin and eosin (HE) staining and Masson staining revealed that PCF exhibited a significant reduction in airway inflammation exudation and collagen fiber production in asthmatic mice. The proteomics analysis revealed that there were 174 proteins exhibiting differential expression in the PCF group compared to the asthma model group, indicating a strong association with negative regulation of the cell cycle and inflammation in the airways of asthmatic individuals. The PCF also exhibited a significant reduction in the protein and mRNA expressions of NLRP3 and caspase-1 in lung tissue (P<0.05). Additionally, it decreased the protein expressions of ASC, p-JNK, MMP-9, E-cadherin, and T-NC while increasing N-cadherin levels (P<0.05). The inflammatory factors IL-1β, IL-18 and TGF-β were reduced (P<0.05). This study revealed that Pingchuan Decoction can inhibit airway remodeling by inhibiting NLRP/IL-1β and JNK pathway activation, and effectively improve airway histological inflammation and remodeling in mice.

Anesthetic sevoflurane (Sev) causes cognitive dysfunction and neuronal death when used as an anesthetic during surgical procedures. Gynosaponin (GpS) was studied for its effects on brain morphology and cognitive behaviors in Sevanesthetized rats. The present study investigated whether GpS has an effect on Sev anesthesia-induced abnormalities in brain morphology and cognitive behaviors, as well as on apoptosis and inflammation of neurons in rats, and delved into the molecular mechanisms. Male Sprague-Dawley rats were induced by 3% Sev anesthesia, and GpS was injected into the rats via the tail vein. The in vitro model of Sev anesthesia was constructed by treating primary rat hippocampal neurons with 4.1% Sev in the presence of GpS (5, 10, and 20 μM). The neuroprotective effects of GpS against Sev-induced cognitive deficits in rats were evaluated using the open field and Morris water maze tests. The apoptosis of hippocampal neurons was observed using hematoxylin-eosin (HE staining and TUNEL assay. Cleaved caspase-3 expression and reactive oxygen species production in rat hippocampal tissue were measured by immunofluorescence. Apoptosis-related proteins and the phosphoinositide 3-kinase (PI3K)/protein kinase B (Akt)/mammalian target of rapamycin (mTOR) pathway were determined via Western blot. Pro-inflammatory factors tumor necrosis factor-α and interleukin-6 were measured via ELISA. Levels of malondialdehyde, superoxide dismutase and glutathione were assayed by commercial kits. Viability and apoptosis of hippocampal neurons were detected by Cell Counting Kit-8 and flow cytometry. Expression of cleaved caspase-3, B-cell leukemia/lymphoma 2 protein (Bcl-2) Bcl-2-associated protein (Bax) was determined by Western blot. GpS significantly reduced Sev-induced decline in short-term memory, learning and cognitive abilities, as well as neuronal degeneration apoptosis and inflammatory responses, GpS also lessened oxidative stress damage, and activated the PI3K/Akt/mTOR pathway (p<0.05). GpS therapy enhanced learning and memory abilities in rats suffering from Sev-induced cognitive deficits. The PI3K/Akt/mTOR pathway inhibitor LY294002 reversed the ameliorative effects of high-dose GpS on cognitive deficits and cell damage in primary hippocampal neurons in Sev anesthetized rats (p<0.05). We conclude that GpS ameliorates Sev-induced neurotoxicity and cognitive dysfunction by modulating the PI3K/Akt/mTOR pathway and alleviating neuronal apoptosis and oxidative stress.

Exposure to maternal diabetes is considered one of the factors during foetal development, leading not only to several complications during pregnancy but also to an increased risk of chronic diseases in later life and exerting a lasting impact on the health of offspring. Animal models play a role in establishing a clear cause-and-effect relationship between prenatal exposure to maternal diabetes and the enduring well-being of offspring, helping to control for variables that could distort the results. This study aims to systematically analysis long-term offspring complications associated with maternal diabetes, drawing insights from both human and animal studies. Analysis expands knowledge about possible new directions of scientific research concerning the prevention and treatment of hyperglycemia in pregnancy through a detailed analysis of the mechanisms of its influence on fetal development.

Melatonin (Mel) has been documented to modulate epithelial-mesenchymal transition (EMT) in cellular systems. The interstitial transformation of renal tubular epithelial cells constitutes a key pathogenic mechanism underlying renal fibrosis. This study aims to elucidate the role of Mel in the EMT process of renal tubular epithelial cells. A rat model of unilateral ureteral obstruction (UUO) was developed through unilateral ureteral ligation, followed by treatment with Mel (5, 10, and 20 mg/kg). Subsequent analyses included histopathological examination, measurement of creatinine and blood urea nitrogen levels, immunofluorescence analysis of fibronectin (FN), and immunohistochemical analysis of alpha-smooth muscle actin (α-SMA). Transforming growth factor-beta 1 (TGF-β1) initiates a fibrotic response in NRK-52E cells, which is subsequently treated with Mel (0. 1, 1, and 10 μmol/L). Evaluates cell viability, migration, and the expression of EMT related proteins, including FN, α-SMA, collagenase I, and E-cadherin. To validate the regulatory interaction between miR-153-3p and Forkhead transcription factor o subfamily member 3A (FOXO3A), cells are transfected with miR-153-3p mimics or siRNA targeting FOXO3A (si-FOXO3A). In results Mel exhibits a dose-dependent capacity to ameliorate renal injury and rectify glomerular structural abnormalities in UUO rat models. In comparison to UUO model rats, melatonin significantly reduced the expression levels of FN and α-SMA. In vitro studies demonstrated that Mel inhibited the activity and migratory behavior of cells, as well as the protein expression levels of FN, α-SMA, and collagenase I in NRK-52E cells. Relative to the control group, UUO rats and TGF-β1-induced NRK-52E cells exhibited elevated expression of miR-153-3p and reduced FOXO3A. In vitro models further revealed that upregulation of miR-153-3p or downregulation of FOXO3A can negate the protective effects of Melatonin on TGF-β1-induced EMT. Concluding, Mel inhibits miR-153-3p, thereby promoting the transcription of FOXO3A in UUO rats, which alleviates renal injury and attenuates TGF-1-induced EMT in cells.

This study aimed to analyze salivary glucose levels and pH in relation to the intensity of dental caries in healthy, nondiabetic adults. The study was conducted on 61 patients who were assigned to individual groups based on dental examinations and the calculation of the decayed, missing, and filled teeth (DMFT) index. Group 1 (control) included 15 caries-free persons (DMFT=0). Group 2 consisted of 15 patients with low caries intensity (DMFT=6.4±1.3). Group 3 comprised 15 patients with moderate caries intensity (DMFT=10.9±1.8). Group 4 included 16 patients with high caries intensity (DMFT=19.2±3.6). Salivary glucose concentration was determined using a quantitative enzymatic method, while salivary pH was measured with a combined electrode. The mean salivary glucose levels in groups 1 and 2 were 2.39±0.62 and 3.28±0.92 mg/dl, respectively. The determined cut-off value for the highest glucose level associated with low caries intensity was 6.64 mg/dl. In contrast, the mean salivary glucose levels in groups 3 and 4 were 7.55±1.16 and 9.9±1.14 mg/dl, respectively, which were significantly higher than those in groups 1 and 2. Regarding salivary pH, the mean values in groups 1 and 2 were 7.19±0.15 and 7.12±0.20, respectively, while in groups 3 and 4, the mean salivary pH were 6.69±0.17 and 6.57±0.19, respectively. These values were significantly lower than the mean values in groups 1 and 2. The data presented here show that increased levels of salivary glucose above 6.64 mg/dl may indicate moderate to high severity of dental caries. Changes in salivary pH may result from the predominant metabolic activity of different species of oral bacteria, including arginolytic, ureolytic or acidogenic bacteria. These findings have important practical implications for monitoring and preventing dental caries.