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0.05).SK & F96365 at the concentration of 5～20 ?mol?L-1 inhibited the Ca2+ influx induced by 1.0 ?mol?L-1 Thapsigargin in a concentration-dependent manner.The inhibitory effect of SK & F96365 on Ca2+ influx was decreased by overexpression of ClC-3 protein.Conclusion ClC-3 chloride channel was involved in the regulation of store-operated Ca2+ entry(SOCE).
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Aim To investigate the method of cell culture for smooth muscle cells from rat cerebral basilar artery and understand cells growth and biological characteristics.Methods The explant attached method was applied for cell culture of rat basilar artery smooth muscle cells(BASMCs).The cultured BASMCs were identified by immunocytochemical staining.The activities of cells were indicated by the dynamic changes of intracellular calcium concentration observed by RF-5 000 fluorospectro-photometer.Results BASMCs grew out of tissue blocks by 5 days,reached confluency could be subcultured after 2 weeks.Cultured cells were identified by intensely positive immunocytochemical staining to smooth muscle actin-specific.Introduction of calcium channel agonists induced significant increase in Fura-2 fluorescence ratio(F340/F380)and cells were in good condiction.Conclusion Explant attached method is simple,efficient and economic.It provides an ideal cell model for the study of pathogenesis of the cerebral vascular diseases.
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Objective:To study the influence of Yizhiping Capsules on hemorrheology of experimental rats with hyperlipoidemia and its embryonic toxicity, teratogenicity. Methods:① The rat experimental hyperlipoidemia models were established by feeding with high lipid diet for 10 days. Meanwhile Yizhiping was taken orally at dosages of 75、150mg?kg -1 , some indices of hemorrheology markers were observed. ②According to the procedure of toxicology, the rat in the experimental groups were administered with Yizhiping capsules by gavage at a dose of 250、500、1000mg?kg -1 respectively, the embryonic toxicity and teratogenicity were observed. Results:① Yizhiping Capsules could significantly raise electrophoresis mobility of red cell of experimental hyperlipoidemia model rats, and lower the rate of red cell deposit and the viscosity of whole blood as well as plasma ( P
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AIM To investigate the role of Trp3 in the Ca 2+ influx induced by ? 1B AR in HEK293 cells and the effect of tyrosine kinase on it. METHODS With lipofect AMINE2000 reagent, hTrp3 cDNA was transfected to HEK293 cells and ? 1B HEK293 cells respectively. The expression of Trp3 was examined by Western blot. With Fura 2/AM spectrophoto fluorometry, Ca 2+ influx was determined. RESULTS HTrp3 was expressed endogenously in HEK293 cells, and the expression increased in hTrp3 transfected cells. Compared with untransfected cells, transfection of hTrp3 cDNA increased Ca 2+ influx induced by ? 1B AR ( P 0 05). 5～30 ?mol?L -1 genistein inhibited Ca 2+ influx induced by ? 1B AR in hTrp3 cDNA transfected cells and the maximum inhibitory rate was (75 2?12 6)% . CONCLUSION Transfection of hTrp3 cDNA increased Ca 2+ influx induced by ? 1B AR in HEK293 cells. This process was regulated by tyrosine kinase.
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AIM To investigate the effect of ClC-3 antisense oligonucleotide on apoptosis induced by thapsigargin in PC12 cells. METHODS Western-blot was performed to detect the protein expression of ClC-3 in PC12 cells. MTT assay was used to measure the effect of ClC-3 antisense oligonucleotide on growth inhibition induced by thapsigargin. The effect of ClC-3 antisense oligonucleotide on apoptosis was studied with the fluorescent microscopy, DNA agarose gel electrophoresis, flow cytometry analysis. RESULTS Compared with control group, transient transfection of PC12 cells with antisense oligonucleotide specific to ClC-3 caused an inhibitory effect on expression of ClC-3 protein in a time-and concentration-dependent manner,whereas the thapsigargin-induced reductions of viability of PC12 cells and apoptosis were markedly enhanced (P
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Aim To investigate the blockade of cadmium on store operated calcium channels and the fluorescence interference of cadmium with Fura-2.Methods PC12 cells were used to determine the intracellular calcium concentration [Ca~(2+)]i indicated by change in Fura-2 fluorescence ratio(F_(340)/F_(380)).Results Introduction of cadmium induced a significant increase in Fura-2 fluorescence ratio following thapsigargin-evoked calcium entry;Besides,cadmium gave rise to a remarkable elevation in Fura-2 fluorescence ratio following breakdown of the plasma membrane by triton,a detergent.The Fura-2 fluorescence was increased in the presence of cadmium and Fura-2/AM,simultaneously in the absence of PC12 cells.Conclusion Cadmium can interfere with the Fura-2 fluorescence.