Hereditas最新文献

Background: Epithelial-mesenchymal transition (EMT) of tubular epithelial cells are one of the major pathological changes of diabetic nephropathy (DN). Cluster of differentiation 248 (CD248) has been reported to be associated with fibrosis after kidney injury. The aim of this study was to investigate the mechanism of CD248 in DN and its targeted compounds.

Materials and methods: Virtual screening, molecular docking and Cellular thermal shift assays were used to explore potential small molecule compounds targeting CD248. In vitro DN model was established by treating human proximal renal tubular epithelial cell line HK-2 with high glucose (HG), and db/db mice were used as the animal model. siRNA transfection was used to knockdown CD248 in HK-2 cells, and HK-2 cells and the animals were treated with veratramine (VER) or neobavaisoflavone (NBIF). qPCR was used to detect the mRNA expression of CD248, tumor necrosis factor-α (TNF-α), interleukin (IL)-6 (IL-6), and IL-1β. Western blot was used to assess protein expression level of CD248, EMT-associated proteins, fibrosis markers, and TGF-β1/Smads pathway-associated proteins. CCK-8 assay and flow cytometry were used to detect cell viability and apoptosis, respectively. Histopathological and various biochemical indicators were used to assess renal injury in animals.

Results: CD248 was significantly up-regulated in HG-induced HK-2 cells. CD248 knockdown inhibited HG-induced cell proliferation inhibition, apoptosis and inflammatory response. HG stimulation significantly reduced the protein expression level of E-cadherin in HK-2 cells, and increased the expression levels of vimentin, α-smooth muscle actin (α-SMA), collagen I, collagen IV, fibronectin, TGF-β1, p-Smad2, p-Smad3, and Smad4, while CD248 knockdown reversed these effects. In addition, VER and neobavaisoflavone were found to bind with CD248, and they inhibited HG-induced apoptosis, inflammation, EMT and extracellular matrix synthesis in HK-2 cells, and ameliorate the renal injury of db/db mice. VER and NBIF also inhibited HG-induced activation of TGF-β1/Smads axis.

Conclusion: CD248 participates in HG-induced EMT of renal tubular epithelial cells and renal fibrosis by regulating TGF-β1/Smads pathway, and VER and NBIF are two potential natural drugs which targets it to ameliorate DN.

Background & objective: MicroRNAs (miRNAs) offer advantages in stability and therapeutic specificity. This study investigated the diagnostic and therapeutic potential of miR-143-5p in sepsis (SP) and SP-associated cardiac dysfunction (CD). ‌Methods. Quantitative Real-Time polymerase chain reaction (qRT-PCR) quantified serum and cellular miR-143-5p levels. Receiver operator characteristic (ROC) curve evaluated miR-143-5p's diagnostic efficacy. Pearson correlation analysis assessed the association between miR-143-5p and SP. Univariate logistic regression identified CD risk factors, with multivariate logistic analysis including significant variables from univariate analysis. Lipopolysaccharide (LPS)-induced macrophage (THP-1) and cardiomyocyte (AC16) models elucidated miR-143-5p mechanisms in SP, with bioinformatics predicting the potential pathways. ‌Results. MiR-143-5p downregulation demonstrated diagnostic value for SP (AUC: 0.897) and SP-CD (AUC: 0.812). MiR-143-5p expression correlated (P < 0.0001) with white blood cell count (WBC, r = -0.680), C-reactive protein (CRP, r = -0.563), procalcitonin (PCT, r = - 0.693), left ventricular ejection fraction (LVEF, r = 0.640), cardiac troponin I (cTnI, r = -0.599), Acute Physiology And Chronic Health Evaluation II (APACHE II, r = -0.695), and Sequential Organ Failure Assessment (SOFA, P < 0.05) scores. MiR-143-5p served as a risk factor for CD in SP (OR: 0.100). MiR-143-5p overexpression reduced M1 polarization and pro-inflammatory cytokines in LPS-treated THP-1. In AC16 cardiomyocytes, it enhanced viability, suppressed apoptosis, and attenuated inflammation. Bioinformatics analysis indicated miR-143-5p was involved in p53 and MAPK signal pathways regulation. ‌Conclusions. MiR-143-5p downregulation showed diagnostic potential for SP and SP with CD, correlating with disease severity and CD risk. Mechanistically, miR-143-5p overexpression mitigated macrophage and cardiomyocyte injury.

Background: Diabetic kidney disease (DKD) is a serious microvascular complication of type 2 diabetes mellitus (T2DM). miR-127-3p is dysregulated in T2DM, but the specific molecular mechanism remains unclear. We aim to probe the diagnostic value of miR-127-3p and its molecular mechanism in T2DM and DKD.

Methods: This study comprised 218 individuals, including 78 patients with T2DM, 72 patients with DKD and 68 healthy controls. All participants underwent fasting peripheral blood collection. In vitro, we simulated a hyperglycemic environment by treating human mesangial cells (HMC) with high-concentration glucose (HG). Subsequently, RT-qPCR was used to detect the levels of miR-127-3p in serum and HMC. Cell viability and inflammatory cytokine (TNF-α, IL-1β and IL-6) levels were assessed using the CCK-8 assay and ELISA, respectively. The dual-luciferase reporter assay validated the target relationship between miR-127-3p and ACO2.

Results: By comparing baseline clinical characteristics, we identified significant differences among the three groups in high density lipoprotein cholesterol (HDL-C), triglycerides (TG), fasting blood glucose (FBG), glycated hemoglobin A1c (HbA1c), blood urea nitrogen (BUN), estimated glomerular filtration rate (eGFR) and albuminuria. Additionally, miR-127-3p was elevated in T2DM and DKD patients. It could distinguish healthy individuals from T2DM or T2DM from DKD. In HG-induced HMC, miR-127-3p inhibitor elevated the cell viability and the levels of SOD while suppressing the levels of MDA. These effects were abolished by ACO2 silencing. Furthermore, downregulated miR-127-3p reduced the levels of TNF-α, IL-1β and IL-6. sh-ACO2 alleviated the inhibitory effects of miR-127-3p.

Conclusions: Upregulated miR-127-3p was involved in the progression of T2DM and DKD. In HG-induced HMC, down-regulated miR-127-3p improved cell viability and suppressed oxidative stress and inflammatory responses by negatively regulating ACO2.

Background: For more than 30 years, Qian Ji Sheng Xue Pian (QJSXP) has been used clinically to treat primary immune thrombocytopenia (ITP) with good documented efficacy. However, nothing is known about its underlying mechanisms, effective components, and possible targets. To employ several methodologies to initially investigate the possible targets and therapeutic mechanisms of QJSXP in the treatment of ITP.

Methods: Liquid chromatography-mass spectrometry (LC-MS) identified the principal chemical elements of QJSXP and assessed its probable active components based on ADME characteristics. The research incorporated multidimensional databases to pinpoint probable targets for the active components. Key pathogenic targets linked to ITP were aggregated from several illness databases, and the STRING and Metascape tools were utilized to examine protein interaction activities and related biological processes. Mendelian randomization (MR) was then utilized to determine beneficial targets for the therapy of ITP. The potential targets, including disease targets and MR-positive targets, were found at the intersection, while risk genes were excluded by heterogeneity, pleiotropy, and Steiger analysis to ascertain the core targets. Molecular docking and molecular dynamics simulations were conducted utilizing Schrodinger and Gromacs software to assess the binding affinity of compound-core targets. The toxicological effects of active molecules targeting critical sites were concurrently anticipated using several toxicity databases.

Results: A total of 67 active components and 352 potential targets were discovered in QJSXP, of which 77 were associated with ITP disease targets. Through MR analysis, a total of 12 core genes were identified. Binding scores below - 4.25 kcal/mol constituted 82.0%; docking scores below - 5 kcal/mol represented 60.1%, with an average binding energy of -5.44 kcal/mol. The majority of targets demonstrated strong binding affinity with the components. Toxicity prediction initially highlighted potential hazards, including hepatotoxicity and nephrotoxicity, establishing a foundation for future clinical surveillance.

Conclusion: This study has preliminarily identified the active constituents, associated pathways, and possible targets of QJSXP in the treatment of ITP, offering insights for additional experimental validation of QJSXP's mechanism of action in ITP.

Background: Complex chromosomal rearrangement (CCR) refers to a structural rearrangement involving at least two chromosomes or a minimum of three breakpoints. CCR may lead to intellectual disability, structural anomalies, infertility, and recurrent miscarriages. Chromosome karyotyping and chromosomal microarray analysis (CMA) are unable to detect complex chromosomal rearrangements. As multiple diagnostic approaches are available in clinical practice for detecting chromosomal structural abnormalities and copy number variations-each with its own advantages and limitations-selecting the appropriate testing method is crucial for effective clinical management. Optical genome mapping (OGM) is an advanced genomic technology that utilizes ultra-long single-molecule analysis to comprehensively detect chromosomal aberrations and structural variants at high resolution.

Material and methods: Amniocentesis was performed for a 36-year-old multipara (advanced maternal age), with subsequent comprehensive fetal genetic analysis including chromosome karyotyping, CMA, and OGM. Family members underwent peripheral blood karyotyping and OGM.

Results: The fetal karyotype derived from amniotic fluid was 46,XN,?ins(18)(q21.2;p11.31p11.2). CMA demonstrated duplications of four segments and a deletion of one segment on chromosome 18. Therefore, OGM was performed on the fetal and family members to further elucidate the chromosomal structure. The fetus has derived CCRs on chromosome 18 of maternal origin. In contrast, both the mother and the second daughter, who carried the identical CCRs, were phenotypically normal.

Conclusion: OGM is of significant importance in the diagnosis and characterization of CCRs. OGM plays a critical role in diagnosing complex chromosomal rearrangements and has proven to be invaluable in clinical utility.