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AIM:To elucidate whether ZFP580 is involved in the cardioprotective effects of hypoxic precondi-tioning (HPC) against hypoxia-reoxygenation (H/R) injury in H9c2 myocardial cells.METHODS: Rat heart-derived H9c2 cells were cultured in DMEM .H/R was induced by incubation under ischemic hypoxia for 3 h and reoxygenation for 2 h.HPC was induced by exposing the H 9c2 cells to 10 min of hypoxia and 20 min of reoxygenation for 3 cycles before H/R treatment.MTT staining and LDH leakage detection were used to evaluate the effects of HPC .Western blotting was used to detect the protein levels of ZFP580, phosphorylated ERK1/2 and cleaved caspased-3.The effects of ZFP580 overexpre-ssion or knockdown on H/R induced apoptosis were determined .RESULTS:The results of MTT staining and LDH leakage detection showed evidence of HPC cytoprotection against H /R-induced cell death in H9c2 cells.ZFP580 protein level and ERK1/2 phosphorylation were significantly increased in the HPC group compared with control group and H /R group. PD98059, an inhibitor of ERK1/2 phosphorylation , significantly suppressed the HPC-induced up-regulation of ZFP580 pro-tein expression.ZFP580 overexpression significantly inhibited apoptosis and caspase-3 activation in H9c2 cells.CON-CLUSION:HPC exhibits cytoprotection against H/R and leads to high level of ZFP 580 protein in H9c2 cells.ZFP580 is regulated by ERK1/2 activation and mediates the anti-apoptotic effect of the ERK1/2 signaling pathway in HPC cytoprotec-tion.
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BACKGROUND: Fibrin is a kind of high polymer materials with biodegradation and good histocompatibility, and is a vector that can promote celland exogenous growth factor release. Fibrin stabilizing factor XIII has been verified to contribute to the migration of undifferentiated mesenchymal stem cels in gel scaffold with high crosslinking, and promote cellproliferation and differentiation. OBJECTIVE:To observe rat mesenchymal stem cellbehavior in a fibrin gel. METHODS:The rat fetal limbs cels was separated under the aseptic condition. The passage 3 cels were seeded in 0, 5, 10 and 20 g/L fibrin gel. cellmorphology was observed by inverted phase microscope and laser scanning confocal microscopy. Alkaline phosphatase activity and calcium deposition were measured respectively using a microplate reader and von Kossa staining. RESULTS AND CONCLUSION: 5 g/L fibrin gel contributed to cellmorphological changes, and 20 g/L fibrin gel contributed to osteogenic differentiation. Compared with the control group, alkaline phosphatase activity was higher in the formulations containing a 20 g/L fibrinogen concentration. Smal mineralization nodules were observed at 21 and 28 days in a formulation containing both 10 and 20 g/L fibrinogen concentration, but no mineralization was detected in the control group. These results indicate that morphology and osteogenic differentiation of rat mesenchymal stem cels depended on the fibrinogen concentration, suggesting that fibrin gel is conducive to osteogenic differentiation of mesenchymal stem cels.
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<p><b>OBJECTIVE</b>To study the effect of pseudolaric acid B (PLAB) on cell proliferation and cycle of human prostate carcinoma DU-145 cells. method: Its inhibitory effect on the cell growth was measured by MTT method. Characteristics of cell death were determined by Hoechest 33342 staining. The cell cycle was detected by flow cytometry. The expressions of cyclin B1, cyclin D1 and CDK1 were detected by Real time-PCR and Western blot, respectively.</p><p><b>RESULT</b>PLAB notably inhibited DU-145 cell growth in a dose- and time dependent manner (P < 0.05). Its IC50 values of PLAB for DU-145 cells for 24, 48 and 72 h were 4.53, 2.39 and 2.08 micromol x L(-1), respectively. Having been treated with 5 micromol x L(-1) PLAB for 24 h, the cells showed such apoptosis characteristics as nuclei chromatin condensation and apoptotic body. With the increase in PLAB concentration, the proportion of G2/M phase cells strikingly increased in a dose- and time dependent manner (P < 0.05), meanwhile cyclin B1 and CDK1 showed over-expressions (P < 0.05), and the cyclin D1 showed under-expression (P < 0.05).</p><p><b>CONCLUSION</b>PLAB can inhibit the growth of DU-145 cells and induce the cell cycle G2/M arrest, accompanied with the over-expression of cyclin B1 and CDK1, which may be related with its regulation cycle-associated protein degradation.</p>
Subject(s)
Humans , Male , Apoptosis , Cell Cycle Checkpoints , Cell Line, Tumor , Cell Proliferation , Diterpenes , Pharmacology , Drugs, Chinese Herbal , Pharmacology , Prostatic Neoplasms , Drug TherapyABSTRACT
BACKGROUND: Fibrin is a natural biodegradable polymer material with good histocompatibility. Basic fibroblast growth factor (bFGF) is an important polypeptide growth factor with mitogenic effect. Fibrin gel (FG) combined with bFGF is conducive to cell migration and promotes proliferation and differentiation. OBJECTIVE: To investigate the effects of fibrin gel combined with bFGF on alkaline phosphatase activity (ALP) in fetal rat limb cells. METHODS: Four groups were included: FG group: fetal rat limb cells were seeded in FG (three-dimensional, 3D) with normal medium; FG + bFGF group: cells were seeded in FG containing 10 μg/L bFGF for 24-hour 3D culture; bFGF group: cells were seeded in 10 μg/L bFGF medium; normal control group: cells were seeded in normal culture medium. Under aseptic condition, rat fetal limb cells were isolated. Passage 3 cells were used for each group above mentioned. ALP activity, mRNA expression and cell morphology were analyzed at 3, 5, 7, 14, 21, and 28 days after culture. RESULTS AND CONCLUSION: The cells in the FG had more processes and formed an interconnected network after 7 days of incubation, while the cells in no gels remained cuboidal or "spindle-shaped". At 7, 14, and 21 days, ALP activity was greater in the FG+ bFGF group than in the other groups (P < 0.05). At each time point, mRNA expression of ALP was greater in the FG+ bFGF group than in the control group (P < 0.01). These findings indicate that FG combined with bFGF is conducive to cell morphogenesis and markedly enhances ALP activity.
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Objective To investigate the expression and correlation of NKG2D and sMICA in lung cancer patients. Methods By collecting 30 lung cancer patients as the test group,and taking 30 healthy volunteers as the contrast group, the expression of NKG2D and sMICA in the two groups were examined separately by FACS and ELISA method. Results The expressions of NKG2D in the two groups were (81.56±8.78) %, (85.63±6.62) %. The lung cancer patients were high remarkable. There was a significant difference between the two groups (P <0.05). The expression of sMICA in the two groups were (354.13 ±80.575) pg/ml,(216.53±48.175) pg/ml. The lung cancer patients were low remarkable. There was a significant difference between the two groups (P <0.01). There was a significant relation between the two groups (r =-0.349, P =0.006). Conclusion The expression of NKG2D and sMICA may provid one of the immune targets for diagnosing that can forecast the immune state and malignant metastasis of the lung cancer patients. The significant relation between NKG2D and sMICA may take on main role in the immune escaping of tumor. It may provide the suitable target of the patients for tumor organisms and immune treatment.