To identify the genes playing a functional role in differentiation of human hepatic progenitor cells to hepatocytes by comparing the gene expression and functional profiles of the two cell types.mRNA was isolated from human fetal hepatic progenitor cells (hFHPCs) and functional hepatocyte-like cells (HLCs) that had differentiated from hFHPCs. Global gene expression profiling was performed on triplicate samples of each cell type. The differential gene expression was analyzed using volcano plot filtering and functional annotation was performed using the Database for Annotation, Visualization, and Integrated Discovery (DAVID).Compared to the hFHPCs, the HLCs had a total of 1878 significantly up-regulated genes and 1441 significantly down-regulated genes. The up-regulated genes included functional groups related to the hexose metabolic process, positive regulation of apoptosis, angiogenesis, regulation of cell motion, and protein amino acid phosphorylation. The down-regulated genes included functional groups related to cell cycle, DNA metabolic process, cytoskeleton organization, regulation cell cycle, and chromosome segregation.Differentiation of HLCs from hFHPCs may involve increased expression of genes related to hepatocyte function and decreased expression of genes related to cell cycle regulation.
Abstract Objective To identify the key drugs of Yangyin Fuzheng Jiedu prescription (YFJP) and investigate their therapeutic effects against hepatocellular carcinoma (HCC) and the potential mechanism using network pharmacology. Methods The H22 tumor‐bearing mouse model was established. Thirty male BALB/c mice were divided randomly into five groups. The mice were orally treated with either disassembled prescriptions of YFJP or saline solution continuously for 14 days. The mice were weighed every 2 days during treatment and the appearance of tumors was observed by photographing. The tumor inhibition rate and the spleen and thymus indexes were calculated. Hematoxylin and eosin and immunohistochemical staining were performed to observe the histological changes and tumor‐infiltrating lymphocytes. Cell apoptosis was determined by terminal deoxynucleotidyl transferase‐mediated dUTP nick‐end labeling staining. The proportion of CD8 + T cells and the expression of programmed cell death protein 1 (PD‐1), T cell immunoglobulin domain and mucin domain‐3 (Tim‐3), and T cell immunoreceptor with Ig and ITIM domains (TIGIT) were analyzed using flow cytometry. The production of serum cytokines was detected using the Milliplex® MAP mouse high sensitivity T cell panel kit. The active components of the key drugs and HCC‐related target proteins were obtained from the corresponding databases. The putative targets for HCC treatment were screened by target mapping, and potential active components were screened by constructing a component‐target network. The interactive targets of putative targets were obtained from the STRING database to construct the protein–protein interaction network. Gene ontology (GO) and Kyoto encyclopedia of genes and genomes pathway enrichment analyses were performed based on potential targets. The gene–gene inner and component‐target‐pathway networks were constructed and analyzed to screen the key targets. Western blotting was used to evaluate the protein expression of the key targets in the tumor‐bearing mouse model. The binding activity of the key targets and compounds was verified by molecular docking. Results Among the three disassembled prescriptions of YFJP, the Fuzheng prescription (FZP) showed significant antitumor effects and inhibited weight loss during the treatment of H22 tumor‐bearing mice. FZP increased the immune organ index and the levels of CD8 + and CD3 + T cells in the spleen and peripheral blood of H22 tumor‐bearing mice. FZP also reduced the expression of PD‐1, TIGIT, and TIM3 in CD8 + T cells and the production of IL‐10, IL‐4, IL‐6, and IL‐1β. Network pharmacology and experimental validation showed that the key targets of FZP in the treatment of HCC were PIK3CA, TP53, MAPK1, MAPK3, and EGFR. The therapeutic effect on HCC was evaluated based on HCC‐related signaling pathways, including the PIK3‐Akt signaling pathway, PD‐L1 expression, and PD‐1 checkpoint pathway in cancer. GO enrichment analysis indicated that FZP positively regulated the molecular functions of transferases and kinases on the cell surface through membrane raft, membrane microarea, and other cell components to inhibit cell death and programmed cell death. Conclusion FZP was found to be the key disassembled prescription of YFJP that exerted antitumor and immunoregulatory effects against HCC. FZP alleviated T cell exhaustion and improved the immunosuppressive microenvironment via HCC‐related targets, pathways, and biological processes.
To clarify the role of Heat shock proteins (HSPs) on the cardiac function during acute myocardial infarction (AMI) after bone marrow cell implantation (BMT), we examined the expression of HSP32 and HSP70 in a rat model of myocardial infarction.Myocardial infarction model was induced in the inbred Lewis rats by left anterior descending artery ligation, and 5 x 10(6) of bone marrow-mononuclear cells (BM-MNCs) were injected into an ischemic zone. On days 1, 3, 7 and 14 post-infarct, the differentiations of transplanted cells and the expressions of HSP32 and HSP70 were determined by immunofluorescence or RT-PCR. The cardiac function was evaluated by echocardiography.Immunofluorescence microscopy of hearts from BMT group revealed that expressions of HSP32 and HSP70 were promoted within cardiomyocytes in the infarction zone and the peri-infarct zone, and expressed within some transplanted bone marrow cells as well. RT-PCR also showed the mRNA expression levels of HSP32 and HSP70 in BMT group were significantly higher than those of the control group, peaked on day 3 post-infarct (5.0-fold and 2.9-fold, respectively, P < 0.01), and then gradually reduced. On day 7 post-infarct, cardiac function (EF and FS) was improved, more than that of the control group (14% and 22%, P < 0.05). On day 14 post-infarct, the specific markers for myocardium or vascular endothelial cells were detected in the transplanted bone marrow cells. The cardiac function was further improved in the BMT group (P < 0.01).At the early phase after BMT, the expressions of HSP32 and HSP70 were upregulated in both transplanted cells and recipient endogenous cardiomyocytes, which improved the acute ischemic cardiac function.
Yangyin Fuzheng Jiedu Prescription (YFJP) is a traditional Chinese medicine (TCM) indicated for the treatment of hepatocellular carcinoma (HCC). Its potential targets and molecular mechanisms are not clear. Therefore, this study intends to explore the molecular mechanism of YFJP based on network pharmacology analysis and in vitro validation.Through univariate and multivariate analyses and survival analysis in HCC patients with or without YFJP treatment we found that drinking alcohol, alfafeto protein ≥ 400 ng/l, baseline portal vein tumor thrombus and total bilirubin level ≥ 18.8 μM) were independent risk factors for poor prognosis, while red blood cell count ≥ 4 × 109/l and TCM treatment were independent protective factors. Besides, YFJP prolonged the cumulative survival of HCC patients. Using online pharmacological methods, we obtained 58 relevant compounds and molecular 53 targets. By using scratch test, Transwell assay, EdU assay, and TUNEL staining, we found that YFJP-containing serum repressed the migration, invasion and proliferation of HCC cells in vitro, and induced cell apoptosis. Moreover, YFJP diminished the gene expression of TP53, CCND1, p-EGFR, EGF, VEGFA, JUN, IL6, COX-2, AKT1, and MAPK1 in HCC cells, but elevated the expression of ESR1 and CASP3.Taken together, results showed that YFJP attenuated HCC progression through mediating effects on HCC-related genes.
AIM To investigate the effects of resveratrol (Res) on the proliferation of VSMCs induced by Ang I and the expression of calmodulin (CaM) and calcineurin (CaN) in the proliferation of VSMCs treated by Ang 1 and to discuss the mechanism. METHODS Rabbit arterial VSMCs were cultured in vitro and VSMCs were identified with the method of immunocytochemistry. A cell proliferating model of VSMCs induced by AngII was established. VSMCs were cultured for 4-8 passages. The experiments were randomly divided into control group, AngII group (0.1 micromol/L) and AngII + Res groups with different concentrations(20, 40, 80, 160) micromol/L. VSMCs proliferation was determined with MTT colorimetric method. CaM was detected with Coomassie brilliant blue method and CaN was determined by enzyme reaction phosphorus measurement. RESULTS Rabbit VSMCs were cultured successfully and could be passaged. After immunocytochemistry staining, all the cells cytoplasm were stained and positive. Cell proliferation, CaM and CaN activities were increased significantly in VSMCs proliferation induced by AngII (P <0.05, P < 0.01). The index of AngII + Res groups were obviously reduced compared with AngII group ( P < 0.01). CONCLUSION The VSMCs proliferation induced by AngII can be inhibited by Res significantly, and the inhibiting mechanism of Res may be related to inhibiting CaM and CaN activities then restraining the proliferation of VSMCs in a dose dependent manner.
Novel simplified Aspirin derivatives were developed, characterized by using IR, 1H NMR, 13C NMR and elemental analysis techniques and evaluated for anticancer activity in human cell lines. The results revealed that most of the compounds exhibited inhibitory effects of growth of cancer cell lines in vitro against T-acute lymphoblastic leukemia cell lines Molt-4, chronic myclogenous leukemia cell lines K-562, acute myelocytic leukemia cells lines HL-60, human breast cancer cell lines MCF-7, human hepatic carcinoma cell lines HepG-2 and human lung cancer cell lines A-549. It was observed that some of these compounds exhibited significant anticancer activity particularly 5i which had stronger antileukemia activity with IC50 values ranging from 11.12 to 19.25 µmol·L-1 against 3 leukemia cells than control fluorouracil, so some of the compounds may constitute a novel class of anticancer medicines, which deserves further study. Keywords: Anticancer activity, Aspirin derivatives, Anti-leukemia activity, Synthesis, MTT, In vitro.
Precise early diagnosis and staging are conducive to improving the prognosis of colorectal cancer (CRC) and gastric cancer (GC) patients. However, due to intrusive inspections and limited sensitivity, the prevailing diagnostic methods impede precisely large-scale screening. In this work, we reported a high-throughput serum metabolic patterns (SMP) screening strategy based on covalent organic frameworks-assisted laser desorption/ionization mass spectrometry (hf-COFsLDI-MS) for early diagnosis and staging of CRC and GC. Notably, 473 high-quality SMP were extracted without any tedious sample pretreatment and coupled with multiple machine learning algorithms; the area under the curve (AUC) value is 0.938 with 96.9% sensitivity for early CRC diagnosis, and the AUC value is 0.974 with 100% sensitivity for early GC diagnosis. Besides, the discrimination of CRC and GC is accomplished with an AUC value of 0.966 for the validation set. Also, the screened-out features were identified by MS/MS experiments, and 8 metabolites were identified as the biomarkers for CRC and GC. Finally, the corresponding disordered metabolic pathways were revealed, and the staging of CRC and GC was completed. This work provides an alternative high-throughput screening strategy for CRC and GC and highlights the potential of metabolic molecular diagnosis in clinical applications.
'/%3e%3cuse xlink:href='%23a' transform='rotate(23.036 .093 25.536)'/%3e%3cuse xlink:href='%23a' transform='rotate(45.87 1.273 16.18)'/%3e%3cuse xlink:href='%23a' transform='rotate(69.945 .996 12.078)'/%3e%3cuse xlink:href='%23a' transform='rotate(20.66 -19.689 31.932)'/%3e%3c/svg%3e)