Cell Biochemistry and Biophysics最新文献

Berberine (BER) and Cyperus rotundus rhizomes extract (CRE) are phytochemicals characterized by broad-spectrum pharmacological activity that could tackle the side effects of conventional mastitis therapies, however, they undergo a modest bioavailability. In the current study, nanoformulations of BER and CRE chitosan hydrogel (BER/CH-NPs, CRE/CH-NPs) were investigated for their antibacterial, antioxidant, anti-inflammatory and anti-apoptotic effects against S. aureus-induced mastitis in a rat model. The experiment was conducted on 80 early lactating female albino rats allocated into 6 groups; control, mastitis, BER/CH-NPs (1 and 0.5 mg), CRE/CH-NPs (0.5 and 0.25 mg), BER/CH-NPs + CRE/CH-NPs (0.5 + 0.25 and 0.25 + 0.125 mg). The nanoparticles were given by oral gavage once every other day from day 2 to day 12 after parturition. On the 13^th day, intra-mammary inoculation with 100 µl of S. aureus suspension containing 2.1 × 10⁸ CFU/ml in all groups except the control group. The results expressed the effect of BER/CH-NPs and CRE/CH-NPs on mammary gland tissue including significantly diminished viable bacterial load as well as attenuated the levels of MPO, MDA, caspase-3 with elevating Nrf2 level, and modulating glutathione redox. Also, the nanoformulations resulted in attenuation of the mRNA expression of TLR2, NOD2, Keap-1 and MAPK signaling pathway additional to the immune reactivity of NF-κB P65 and p-ERK as well as the preservation of the regular alveolar architecture. The supplementation of the berberine and Cyperus rotundus extract nanoformulations could be a prospective protective approach against Staphylococcal mastitis via their antibacterial, antioxidant, antiapoptotic, anti-inflammatory and modulation of MAPK signaling pathway.

Apical periodontitis is an inflammatory disease caused by bacterial infection in the root canal that spreads to the apical periodontal tissues, resulting in bone resorption around the root apex as the disease progresses. Vascular endothelial growth factor (VEGF), a growth factor involved in angiogenesis, plays an important role in bone remodeling. We reported that caffeic acid phenethyl ester (CAPE), a bioactive substance of propolis, induces VEGF in odontoblast-like cells and dental pulp cells. However, the effects of CAPE on bone tissues remain unclear. This study was aimed to investigate the effects of CAPE on MC3T3-E1 cells, mice preosteoblast line. As a result, CAPE up-regulated the production of VEGF and induced the phosphorylation of extracellular signal-regulated kinases (ERK), p38 mitogen-activated protein kinase (MAPK), and stress-activated protein kinase/c-Jun N-terminal kinase (SAP/JNK) in MC3T3-E1 cells. Furthermore, CAPE increased the expression of factors involved in osteoblast differentiation, runt-related transcription factor 2 (Runx2), Osterix, and Wnt5a/b in MC3T3-E1 cells. In this study, we show that CAPE could induce bone repair-related factors in MC3T3-E1 cells.

Glutathione (GSH) is a master antioxidant that counters oxidative stress. Clinical studies have confirmed significant depletion of GSH in the hippocampus and the substantia nigra as an early diagnostic biomarker for Alzheimer's disease (AD) and Parkinson disease (PD), respectively. External agents like anesthetics (inhaled and intravenous) have a different impact on GSH. There is significant depletion of the serum GSH peroxidase level after surgery with isoflurane anesthesia that is not found in patients administered intravenous propofol. The objective of this study is to evaluate the GSH level associated with isoflurane in vitro phantom model using non-invasive magnetic resonance (MR) spectroscopy and to detect residual isoflurane in a solution. MRS data was generated utilizing a 3T MR scanner (Prisma, Siemens) equipped with a 64-channel ¹H head coil and dual tune (¹⁹F/¹H) head coil. The GSH data acquisition was performed using the MEGA-PRESS pulse sequence using experimental parameters: ON = 4.40 ppm, OFF = 5.00 ppm, TE = 120 ms, TR = 2500 ms, voxel size = 25 × 25 × 25 mm and average = 32. Isoflurane was detected using ¹⁹F MRS studies using ¹⁹F/¹H head coil. GSH data was processed using KALPANA package and ¹⁹F data was processed using Siemens package. The GSH peak area (without isoflurane) in a phosphate-buffered solution (PBS) solution (control) showed a slow decline over time due to natural oxidation of GSH to dimeric glutathione (GSSG). On the contrary, the GSH peak area in similar model is reduced significantly (p = 0.016) due to isoflurane induced oxidation of GSH to GSSG compared to control. We also report a concise general method for data generation and processing of ¹H MRS data for GSH as well as ¹⁹F monitoring platform using ¹⁹F MR spectroscopy. This is the first report wherein both ¹H and ¹⁹F spectroscopy are applied to generate MRS data along with a unique data processing method. This method is highly sensitive and specifically detects GSH without ambiguity as well as isoflurane due to the unique chemical shift patterns of CF₃ and CHF₂ moieties. This non-invasive MRS approach is developed to monitor GSH-isoflurane interaction leading to oxidative stress and this approach can be extended for other inhaled anesthetics. This methodology using non-invasive ¹⁹F MR spectroscopy needs further development for future clinical studies.

To investigate the regulatory mechanism of indoleamine 2, 3-dioxygenase (IDO) in T lymphocyte differentiation and its role in promoting the growth of gastric cancer (GC) cells through the PI3K/Akt/mTOR pathway. GC cell lines (MFC and NCI-N87) and PBMC cells were co-cultured and IDO inhibitor 1-methyl-tryptophan (1-MT) was added. The proliferation was detected by CCK-8, the apoptosis was detected by flow cytometry, and the contents of TNF-α, IL-1β, IL-6, IL-8, and INF-γ were detected by ELISA. The expression levels of PI3K, p-PI3K, Akt, p-Akt, mTOR, and p-mTOR were tested using Western blot, and the proportion of CD4⁺/CD8⁺, CD4⁺CD25⁺Foxp3⁺Treg cells was detected by flow cytometry. C57BL/6 mice were used to establish the MFC GC mouse model and treated with 1-MT. The changes in body weight and tumor diameter were measured. Ki-67, CD4⁺, CD8⁺, and CD25⁺ expressions were detected by immunohistochemistry. IDO promoted the proliferation of MFC and NCI-N87 cells, inhibited apoptosis, and decreased the levels of TNF-α, IL-1β, IL-6, IL-8, and INF-γ in the supernatant after co-culture with BPMC. The expressions of p-AKT, p-mTOR, and p-PI3K increased after 1-MT treatment. The proportion of CD4⁺/CD8⁺ cells was increased and the proportion of Treg cells was decreased in PBMC cells after the addition of 1-MT. Overexpression of IDO suppressed T cells differentiation by inhibiting the PI3K/Akt/mTOR pathway. In vivo, 1-MT treatment reduced the tumor size and weight, increased CD4⁺ and CD8⁺ positive area proportion, and decreased Ki-67 and CD25⁺ positive area proportion. Co-culture of GC cells and immune cells promotes the proliferation of GC cells and inhibits apoptosis, which can be reversed by 1-MT. IDO may suppress the proliferation of T lymphocyte through inhibiting the PI3K/Akt/mTOR signaling pathway. This provides new evidence for the potential of exploiting IDO inhibitors for GC treatment.

Lung cancer (LC) accounts for approximately 25% of all cancer cases, with 80-85% of these being non-small cell lung cancer (NSCLC). VS-5584 is a novel anti-cancer agent that specifically inhibits mTORC1/2 and class I PI3K isoforms. There is cross-talk between the PI3K-Akt-mTOR and WNT signaling pathways that are abnormally activated in NSCLC. In this study, we aimed to evaluate the anti-cancer effects of VS-5584 on A549 lung adenocarcinoma cells and changes in WNT signaling gene expression in vitro, while also correlating differentially expressed genes in silico. The effect of VS-5584 on A549 cell viability was assessed by the MTT assay. Apoptosis and cell cycle profiles were analyzed by flow cytometry, while WNT signaling gene expression was measured by quantitative RT-PCR. Differentially expressed genes (DEGs) in the TCGA LUAD and LUSC datasets were identified using the GEPIA2 platform. VS-5584 treatment induced apoptosis and caused cell cycle arrest at the G0/G1 phase in A549 cells. The mRNA expression levels of WNT signaling genes significantly decreased in treated cells. The expression of some upregulated DEGs in the datasets decreased in A549 cells treated with VS-5584. VS-5584 shows promise as an anti-cancer agent in the treatment of NSCLC by downregulating the expression of WNT signaling genes.

Traumatic brain injury (TBI) is a common traumatic event that imposes a significant burden on families and society. Lipocalin (LCN) is a class of multifunctional secreted lipoprotein molecules. This study aimed to explore the role and possible mechanism of LCN2 in TBI. A rat model of TBI was constructed and adeno-associated virus-coated shRNA-LCN2 was used to silence LCN2 expression. The modified neurological severity score (mNSS), learning and memory ability, pathological injury of brain tissue, number of neurons, and expression of neurotrophic factors were analyzed, and the expression of inflammatory factors, M1/M2 polarization of microglia, and p38MAPK-PGC-1α-PPARγ pathway after LCN2 silencing were further detected. Results found that LCN2 was highly expressed in the brain tissue of TBI rats, and there were obvious learning and cognitive impairments and pathological injury of brain tissue. After silencing LCN2, the mNSS was further increased, and the learning and cognitive ability was weakened. Similarly, silencing LCN2 increased the brain tissue water content, aggravated the histopathology degree, decreased the number of surviving neurons, and reduced the expression of neurotrophic factors in TBI model rats. In addition, the expression of M1 proinflammatory cytokines and polarization markers in microglia of TBI was increased, and the expression of M2 cytokines and markers was decreased after silencing LCN2. Silencing LCN2 also inhibited the activation of the p38MAPK-PGC-1α-PPARγ pathway. In conclusion, LCN2 was released by surviving neurons after TBI, and the increased LCN2 activated the p38MAPK-PGC-1α-PPARγ pathway, which promoted M2 polarization of microglia, and secreted neurotrophic factors, thereby alleviating secondary brain injury.

Periodontitis, a chronic inflammatory condition, often results in gum tissue damage and can lead to tooth loss. This study explores the role of methyltransferase-like 3 (METTL3) in promoting osteogenic differentiation of human periodontal ligament stem cells (hPDLSCs) within an inflammatory microenvironment. An inflammatory environment was simulated in hPDLSCs using lipopolysaccharide (LPS). Both adipogenic and osteogenic differentiation capacities of hPDLSCs were assessed. In LPS-treated hPDLSCs, METTL3 was overexpressed, and alkaline phosphatase (ALP) staining was performed alongside measurements of ALP activity, pro-inflammatory cytokines, METTL3, miR-141-3p, pri-miR-141, Zinc finger E-box binding homeobox 1 (ZEB1), runt-related transcription factor 2 (RUNX2), osteocalcin (OCN). N6-methyladenosine (m6A) and pri-miR-141 levels were quantified, and the binding of miR-141-3p to ZEB1 was analyzed. The results demonstrated that osteogenic differentiation in hPDLSCs was diminished under inflammatory conditions, coinciding with downregulated METTL3 expression. However, METTL3 overexpression enhanced osteogenic differentiation. METTL3 facilitated the conversion of pri-miR-141 into miR-141-3p via m6A modification, resulting in increased miR-141-3p levels, which in turn suppressed ZEB1 expression. Inhibition of miR-141-3p or overexpression of ZEB1 partially counteracted the positive effects of METTL3 on osteogenic differentiation. In conclusion, these findings suggest that METTL3-mediated m6A modification promotes osteogenic differentiation of hPDLSCs within an inflammatory microenvironment through the miR-141-3p/ZEB1 axis.

Oral submucous fibrosis (OSF) is a precancerous lesion of the oral cavity. Areca nut consumption can cause OSF through sustained activation of buccal mucosal fibroblasts (BMFs). This study explored the effect of curcumin on arecoline-induced BMF activation and its mechanism of action. BMFs were isolated and identified by immunofluorescence detection of fibroblast surface markers vimentin and S100A4. After transfection with FOSL1- or MAPK8-related vectors, BMFs were activated by arecoline and treated with curcumin. Scratch and transwell assays were performed to detect cell migration. ChIP and luciferase reporter assays were conducted to detect the binding of FOSL1 to the MAPK8 promoter. RT-qPCR was used to detect FOSL1 and MAPK8 mRNA expression. Western blotting was used to detect FOSL1, MAPK8, COL1A1, α-SMA, Smad2, and p-Smad2 proteins. Curcumin treatment inhibited arecoline-induced fibroblast migration, reduced the expression of myofibroblast markers COL1A1, α-SMA, and p-Smad2, and downregulated the expression of FOSL1 and MAPK8. FOSL1 or MAPK8 overexpression enhanced migration and increased COL1A1, α-SMA, and p-Smad2 expression in curcumin-treated cells. FOSL1 bound to the MAPK8 promoter and promoted MAPK8 expression. Simultaneous FOSL1 overexpression and MAPK8 knockdown, compared to FOSL1 overexpression, reduced cell migration and inhibited COL1A1, α-SMA, and p-Smad2 expression. In conclusion, curcumin targets FOSL1 to reduce MAPK8 expression, thereby suppressing arecoline-induced fibroblast activation.

Breast cancer is a disease that seriously endangers the health of women. However, it is difficult to treat due to the emergence of metastasis and drug resistance. Exploring the metastasis mechanism of breast cancer is helpful to aim for the appropriate target. The epithelial-mesenchymal transition (EMT) is an important mechanism of breast cancer metastasis. Sodium channel 1.5(Na_V1.5) and the GTPase Rac1 are factors related to the degree of malignancy of breast tumors. The expression of Na_V1.5 and the activation of Rac1 are both involved in EMT. In addition, Na_V1.5 can change the plasma membrane potential (Vm) by promoting the inflow of Na⁺ to depolarize the cell membrane, induce the activation of Rac1 and produce a cascade of reactions that lead to EMT in breast cancer cells; this sequence of events further induces the movement, migration and invasion of tumor cells and affects the prognosis of breast cancer patients. In this paper, the roles of Na_V1.5 and Rac1 in EMT-mediated breast cancer progression were reviewed.