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Objective To investigate the effect of serum-deprivation on the changes of [ Ca2+ ] i and the protein release of S100A13 and fibroblast growth factor 1 ( FGF-1 ) in thyroid cancer TT cell,and to reveal the role of Ca2+in the protein release of S100Al3 and FGF-1.Methods The protein expressions of FGF-1 and S100A13 in TT cells under serum-deprivation were detected by Western blot.The released FGF-1 protein from TT cells in the supernatant fluid was detected by ELISA.Realtime dynamic examinations on the change of 1 h [ Ca2+ ] i in TT cells under serum-deprivation were detected by confocal laser scanning microscopy.Then,the effect of EGTA( 2.5 mmol/ L),BAPTA-AM (2.5 μmol/L)on distributions of the fluorescence of S100AI 3 and FGF-1 in TT cells under serumdeprivation for6 h were detected by indirect immunofluorescence.Results The expressions of FGF-1 and S100A13 in TT cells after serum-deprivation for4 h and 6 h were reduced( P＜0.05 or P＜0.01 ),but the released FGF-1 protein from TT cells in the supernatant fluid was elevated ( P＜0.05 or P＜0.01).Confocal laser scanning of Ca2+ imaging indicated that [ Ca2+ ] i of serum-deprivation TT cells maintained the relative stabilization within 23 win,but the rapid rise of [ Ca2+ ] i achieved peak value 1.6 μmol/L after 30 min,and remained stable for about 17 win,and thereafter 40 win slowly dropped to a low level From 40 win to 60 win the [ Ca2+ ] i was about 0.3-0.6 μ mol/L.The average [ Ca2+ ] i was higher than that in normal group,EGTA group,and BAPTA-AM group within 1 h.The protein expressions of S100A13 and FGF-1 did not drop obviously in EGTA group and BAPTA-AM group.Conlusion The release of S100A13 and FGF-1 from TT cell under serum-deprivation is possibly related with the change of [ Ca2+ ]i.Both Ca2+-chelating agents EGTA and BA PTA-AM are able to inhibit the rise of [ Ca2+ ] i and release of S100A 13 and FGF-1 from TT cells under serum-deprivation.
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AIM To observe protective effects of doxepin on focal cerebral ischemia-reperfusion(I-R)injured rats. METHODS 120 rats were randomly divided into ischemia-reperfusion group,doxepin, nimodipine group and sham-operated group respectively, ischemia-reperfusion model was made by using the suture emboli method, the content of NO and MDA, activities of NOS and SOD in the cerebral tissues and the intracellular [Ca 2+ ] i in the cerebral cortex were determined respectively. RESULTS The content of NO, MDA and [Ca 2+ ] i and the activities of NOS decreased and the activities of SOD increased significantly in the doxepin and nimodipine group compared with ischemia-reperfusion group respectively(P
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AIM: To investigate the influences of nimodipine on HCT cells proliferation and explore the mechanism. METHODS: HCT cells were treated with different concentrations of nimodipine, and its proliferation was inspected by MTT assay. The apex areas of sub diploid were measured by Flow Cytometry and the DNA ladder were found by agarose gel electrophoresis. The characteristic changes in morphology were observed under the light microscopy. The cellular distribution and concentration of calcium were studied by using the laser confocus scanning microscopy. RESULTS: The growth of HCT cells was inhibited by different concentrations of nimodipine. Data from Flow Cytometry showed the apex areas of sub diploid enlarge in the drug treating group, suggesting that the number of apoptosis cells increased in dose dependent manner. Gel electrophoresis displayed DNA cleavage pattern typical of apoptosis: DNA ladder in high dose nimodipine treated HCT cells. The light microscopy presented cellular morphological changes: cell membrane blebs, the cytoplasm and nuclear chromatin condensation. The concentration of cytosolic free calcium increased when treated with nimodipine showed by the laser confocus scanning microscopy. CONCLUSION: Nimodipine can cause inhibition of the cell proliferation of HCT cells. The mechanism of inhibition might involve the cell apoptosis.
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Object To study the effect of emodin, the active principle in rhubarb on the secretion of TNF ?, IL 1, IL 6, and [Ca 2+ ] i by rat peritoneal macrophage under different conditions Methods Hypernomic inflammation models of isolated rat peritoneal macrophage were prepared by lipopolysaccharide excitation and the levels of [Ca 2+ ] i and the proinflammatory cytokines TNF ?, IL 1 and IL 6 secreated determined by fluorescence spectrophotometry and bioassay Results Emodin showed a dual regulatory effect on the secretion of proinflammatory cytokines and [Ca 2+ ] i Conclusion It seemed that emodin has a biphasic immuno regulatory effect
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AIM To examine the change of intracelluar Ca 2+ level regulated by EPA in bovine aorta endothelium of different stages of culture and determine the time that cultured endothelium loses functional EPA. METHODS As fluorescence probe, Fluo 3 and Fura 2 were used in assaying [Ca 2+ ] i in endothelium by confocal microscopy and fluorophotometry. RESULTS Ach could activate EPA and elicite the [Ca 2+ ] i change of endothelium in certain periods of culture, especially in primary stage of cul ture. The rises of [Ca 2+ ] i exhibited as oscillation characterized by iso timing and immediate tolerance. The functional EPA was lost in passage 13. CONCLUSION EPA can be activated by Ach, which mediated the rise of intracelluar Ca 2+ in bovine aorta endothelia cell. This character of EPA is lost during passaging.
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AIM To study the effects of propofol on membrane fluidity and intracellular free Ca 2+ concentration ([Ca 2+ ] i ) in PC12 cells and discuss its relevant mechanism. METHODS PC12 cell lines were divided into seven groups: control, solvent and propofols(1,3,10,30,100 mg?L -1 ). Fluorescence depolarization method was used to measure dynamically microviscosity in PC12 cells and [Ca 2+ ] i was detected using calcium fluorescentprobe Fluo 3/AM and a laser scanning confocal microscope. RESULTS ①Acute administration of various doses of propofol induced a significant decrease of microviscosity in PC12 cells dose dependenty. ② Solvent, propofol at dose of 10 mg?L -1 had no effect on [Ca 2+ ] i in PC12 cells, however, after 30 and 100 mg?L -1 administration, [Ca 2+ ] i increased markedly at 20～30 seconds (increase percentage were 119% and 140% respectively) and then recovered to their pre administration levels within 50 seconds. CONCLUSION The propofol can significantly increase membrane fluidity in PC12 cells in a dose dependent manner and elevate [Ca 2+ ] i in PC12 cells at doses of 30 and 100 mg?L -1 . These changes are consistent with each other and related closely with anesthetic effect of propofol.