ABSTRACT
BACKGROUND: In recent years, bone marrow mesenchymal stem cells have been used in clinical treatment of heart injury caused by permanent cardiomyocytopenia. However, the best inducer to promote the proliferation and differentiation of bone marrow mesenchymal stem cells into cardiomyocytes is still in the research and selection. OBJECTIVE: To investigate the protective effect of salvianolic acid B on oxidative stress injury and cardiomyocyte differentiation of bone marrow mesenchymal stem cells. METHODS: Bone marrow mesenchymal stem cells were used as study objects to establish oxidative stress injury model by 300 μmol/L H2O2 for 24 hours. Simultaneously, bone marrow mesenchymal stem cells were treated with salvianolic acid B for 24 hours. The contents of lactate dehydrogenase and creatine kinase in the supernatant and malondialdehyde and superoxide dismutase in the cell lysate were determined in each group using the kit. Western blot assay was used to detect the expression of nuclear factor E2 related factor 2 (Nrf2), Keap1, Bcl-2 and Bax in bone marrow mesenchymal stem cells. RT-PCT was used to detect cardiomyocyte differentiation-related markers 14 days after intervention. RESULTS AND CONCLUSION: (1) Compared with the model group, the levels of lactate dehydrogenase and creatine kinase in the supernatant were significantly decreased in the salvianolic acid B group (P < 0.05). (2) Compared with the model group, the level of malondialdehyde in cell lysate was decreased, and the level of superoxide dismutase was increased in the salvianolic acid B group (P < 0.05). (3) Compared with the model group, expression of Nrf2 protein in cells was significantly increased, and the expression of Keap1 protein was significantly decreased in the salvianolic acid B group. (4) The expression of Bcl-2 in salvianolic acid B group was significantly higher than that in model group, and the expression of Bax in salvianolic acid B group was significantly lower than that in model group. (5) Compared with the model group, the expression of GATA4 and cTnT mRNA was significantly increased in the salvianolic acid B group. (6) Salvianolic acid B can induce the expression of proteins that activate Nrf2-ARE signaling pathway in bone marrow mesenchymal stem cells for anti-oxidation, and salvianolic acid B can inhibit apoptosis and improve the ability of bone marrow mesenchymal stem cells to differentiate into cardiomyocytes.
ABSTRACT
To construct soluble TNF related apoptosis inducing ligand (TRAIL) expression system and investigate the effect of the expression product on tumor cell. It may provide valuable information for research into the immune system of the finless porpoise. The full-length cDNA of TRAIL (designated fTRAIL) was cloned from the total RNA of the finless porpoises blood using RT-PCR techniques and then the extracellular soluble fragments of fTRAIL (designated fsTRAIL) was ligated into pET43.1a. Recombinant soluble fTRAIL (pET43.1a-fsTRAIL) fused with Nus-his tag was efficiently expressed in Escherichia coli BL21 (DE3) and the Nus-His-fsTRAIL protein was purified. The expression of Nus-His-fsTRAIL was verified by Western blotting. In vitro, the effects of the purified Nus-His-fsTRAIL protein on Jurkat and HeLa cells were etected by the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrasodium bromide (MTT) assay, TrypanBlue and Flow Cytometry analysis. The expression system pET43.1a-fsTRAIL was constructed and Nus-His-fsTRAIL protein was expressed successfully. In vitro, the Nus-His-fsTRAIL protein was able to inhibit the proliferation and induce apoptosis of Jurkat and HeLa cells in a dose-dependent manner. The Nus-His-fsTRAIL protein has anti-tumor activity against Jurkat and HeLa cells in vitro.
Subject(s)
Animals , Humans , Apoptosis , Blotting, Western , Cloning, Molecular , DNA, Complementary , Escherichia coli , HeLa Cells , Jurkat Cells , Porpoises , TNF-Related Apoptosis-Inducing LigandABSTRACT
Objective To probe the clinical values of the human serum glycoprotein profiles for the diagnosis of common gynecological tumors. Methods A total of 123 clinical serum samples which included 31 breast cancer, 24 cervical cancer, 19 ovarian cancer and 49 healthy individuals were collected. A lectin microarray consisting of 15 lectins with different glycan binding specificities was used to determine the glycoprotein profiles of serum sam-ples. Stepwise discrimination analysis method was adopted to establish function model of clinical serum samples classification with SPSS 15. 0 software. Results Two grades of diagnostic discrimination function models were es-tablished. The first grade discrimination function could differentiate gynecological tumors from healthy individuals, the diagnostic accuracy rates of retrospective inspection were 85. 7% and 83. 8% respectively, and the total diag-nostic accuracy rate was 84.6%. The second grade discrimination function was used to differentiate breast tumor, cervical tumor and ovarian tumor, the diagnostic accuracy rates of retrospective inspection were 96.8%,75.0%and 78.9% respectively, and the total diagnostic accuracy rate was 85.1%. Conclusion The human serum gly-coprotein profiles are associated with gynecological tumors, and the established discrimination function models based on lectin microarray data have a helpful reference value for the clinical diagnosis of gynecological tumors.