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Journal of Huazhong University of Science and Technology(Medical Sciences) 2004年第5期24卷 424-426页

作者：张蕲胡波孙圣刚邓学军梅元武童萼塘 Department of Phathophysiology School of Basic Medical Sciences Tongji Medical College Huazhong University of Science and Technology Wuhan 430030 China Department of Neurology Union Hospital Tongji Medical College Huazhong University of Science and Technology Wuhan 430030 China

By using Fura-2/AM, the effects of magnesium (Mg 2+) on the glutamate-induced increase of intracellular free calcium ([Ca 2+]i) in the cultured hippocampal neurons and the features were investigated by integrated photoelectric detecting system. The experiments were designed to three groups (The drug was spit to the cells for 20 s): Group A receiving 1×10 —5 mol/L glutamate; Group B receiving 1×10 —5 mol/L glutamate and1×10 —5 mol/L Mg 2+ simultaneously; Group C receiving 1×10 —5 mol/L glutamate again after [Ca 2+]i in group B back to the baseline. The results showed that in group A, [Ca 2+]i was obviously increased. In group B, the changes in [Ca 2+]i and the peak value were significantly decreased. Moreover, the elevation of Phase 1 was slowed down and Phase 2 was shortened to some extent, and the plateau phase between them was relatively prolonged. In group C, calcium oscillation similar to that in group A occurred, but both the Phase 1 and Phase 2 were shortened and the △[Ca 2+]i was slightly decreased. It was suggested that Mg 2+ could quickly inhibit the rise of [Ca 2+]i induced by glutamate in the cultured hippocampal neurons in rats.

关键词： magnesium glutamate intracellular free calcium

Both Hypoxic Endothelial Cell Conditioned Medium and Hypoxia Elevate intracellular free calcium in Pulmonary Artery Smooth Muscle Cells

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Journal of Huazhong University of Science and Technology(Medical Sciences) 1994年第4期14卷 200-203页

作者：胡清华王迪浔 Department of Pathophysiology Tongji Medical University Wuhan

By using Ca2+ -sensitive fluorescent probe, Fura-2 , the effects of endothelial cell-conditioned medium and hypoxia on intracellular free calcium ( [Ca2+]i) in cultured pulmonary artery smooth muscle cell (PASMC) were studied. Normoxic porcine pulmonary artery endothelial cell-conditioned medium (NPAECCM) obviously elevated [Ca2+]i in PASMC,whereas the hypoxic porcine pulmonary artery endothelial cell conditioned medium (HPAECCM)significantly elevated [Ca2+]i in PASMC much more than NPAECCM. Both the effects of NPAECCM and HPAECCM were dependent on the cultured endothelial cell extracellular calcium concentrations, ranged from 1.8 mmol/L to 2. 4 mmol/***, hypoxia directly increased, which was partially inhibited by verapamil,[Ca2+]i in PASMC through Ca2+ influx *** data suggest that the augmented regulation of endothelial cell on PASMC via Ca2+ second messenger system and the hypoxia-induced Ca2+ influx into PASMC,particularly the former, may be components of mechanisms underlying hypoxic pulmonary vasoconstriction and chronic pulmonary hypertension.

关键词： intracellular free calcium hypoxia,endothelial cell smooth muscle cell

Effects of Total Flavonoids ofHippophae *** intracellular free calciumin Cultured Vascular Smooth Muscle Cells of Spontaneously Hypertensive Rats and Wistar-Kyoto Rats

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Chinese Journal of Integrative Medicine 2005年第4期11卷 287-292页

作者：朱福黄波胡春燕蒋庆渊卢振国陆铭王美华龚敏乔春萍陈维黄盼华 Cardiovascular Depart ment Nanhui District CentralHospital Shanghai 201300 Cardiovascular Depart-ment Shanghai Renhe Hospital Internal Medicine De-part ment Longhua Hospital Shanghai University of Tradi-tional Chinese Medicine

Objective: To explore the effects of total flavonoids of Hippophae rhamnoides L. (TFH), quercetin (Que) and isorhamnetin (Isor) on the intracellular free calcium (Ca~ 2+ _ i) in vascular smooth muscle cells (VSMC) of spontaneously hypertensive rats (SHR) and Wistar-Kyoto rats (WKY).Methods: Fluo 3-acetoxymethylester(Fluo-3/AM) was used to observe the effects of TFH (100mg/L) and its essential monomers, namely Que (10~ -4 mol/L) and Isor (10~ -4 mol/L) on changes of Ca~ 2+ _ i in cultured SHR and WKY VSMC (abbr. to Ca-SHR & Ca-WKY) following exposure to high K~+, norepinephrine (NE) and angiotensin Ⅱ(AngⅡ), and to compare with the effects of verapamil (Ver). Results: (1) TFH, Que and Isor had inhibitory effects on resting Ca-SHR (P0.05). (2) High K~+ could increase Ca-SHR more significantly than Ca-WKY (Pcalcium channels (VDC) and receptor-operated calcium channels (ROC) in physiological or pathological state, which may be one of the important mechanisms of their hypotensive and protective effects on target organs in patients with hypertension.

关键词： total flavonoids of Hippophae rhamnoides L. spontaneously hypertensive rats Wistar-Kyoto rats vascular smooth muscle cells intracellular free calcium

High concentration of calcium ions in Golgi apparatus

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Cell Research 1994年第1期4卷 97-108页

作者： XUE SHAOBAI M. RoBERT NICOUD JIE CUI D.J.ARNDT JOVIN(Depariment of Biology, Beijing Normal University, Beijing 100875, China)(Max-Planck-Institute fur Biophysikalische Chemie,Gottingen, Germany) Department of Biology Beijing Normal University Beijing China Max-Planck-Institute fSr Biophysikalische Chemie GSttingen Germany

The interphase NIH3T3 cells were vitally fluorescentstained with calcium indicator fluo-3 and Glogi probe C6NBD-ceramide, and then the single cells were examined by laser scanning confocal microscopy (LSCFM) for subcellular distributions of Ca2+ and the location of Golgi apparatus. In these cells, the intracellular Ca2+ were found to be highly concentrated in the Golgi apparatus. The changes of distribution of cytosolic high Ca2+ region and the Golgi apparatus coincided with the cell cycle *** calcium free medium, when the plasma membrane of the cells which had been loaded with fluo-3/AM were permeated by digitonin, the fluorescence of the Golgi region decreased far less than that of the cytosol. Our results indicated that the Glogi lumen retained significantly high concentration of free calcium.

关键词： intracellular free calcium fluo-3/AM Golgi apparatus C6-NBD-ceramide laser scanning confocal microscopy intracellular calcium store

Effect and Mechanism of Epidermal Growth Factor on Proliferation of GL15 Gliomas Cell Line

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Journal of Huazhong University of Science and Technology(Medical Sciences) 2006年第5期26卷 604-606页

作者：王和平郭东生叶飞席桂发王宝峰陈坚雷霆 Department of Neurosurgery Tongji HospitalTongJi Medical CollegeHuazhong University of Science and TechnologyWuhan 430030China

The effects of epidermal growth factor （EGF） on proliferation of G15 glioma cells and the possible mechanisms were investigated. GFAP and EGFR expression was detected by immunohistochemical method. After the cells were treated with EGF at different concentrations, cell count method was used to determine the proliferation of glioma cells, cell cycle and apoptosis were analyzed by flow cytometry （FCM）, and laser scan confocal microscope （LSCM） was used to measure the cytoplasmic free calcium. The results showed that GFAP was diffusedly expressed in GLI5 cells and EGFR was over-expressed. EGF at doses of ≤ 1 ng/mL could significantly stimulate cell proliferation, cells in phase G0/G1 decreased, and those in phase S increased. EGF at doses of 10 and 100ng/ml could inhibit the cell proliferation significantly, and the apoptosis ratio in high dose of EGF group was higher than in control group. EGF could significantly induce a quick rise of intracellular free calcium, but the peak value of intracellular free calcium activated by high dose of EGF was higher than by low dose of EGF. It was suggested that EGF had a dual effect on gliomas： low dose of EGF could stimulate the cell proliferation of gliomas, but high dose of EGF could induce the cell apoptosis and inhibit the proliferation of gliomas, which might be contributed to the difference of intracellular free calcium.

关键词： epidermal growth factor GL15 cell line proliferation intracellular free calcium apoptosis

Set up Alzheimer＇s Disease Cell Apoptosis Model with PC-12 Cell Induced by Aβ25-35

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Journal of Nanjing Medical University 2003年第5期17卷 208-214,260页

作者：王亚利王中卫宋潍杨林 DepartmentofOncology theAffiliatedHospitalofMedicalCollegeofQingdaoUniversityQingdao266003 DepartmentofOncology theSecondHospitalofXi’anJiaotongUniversityXi’an710061P.R.China

Objective: To prepare an apoptosis cell model of Alzheimer Disease (AD) by PC-12 cells treated with β-amyloid protein (Aβ). Methods: PC-12 cells were incubated with different concentrations of Aβ25-35 for different duration in vitro. The cell viability was detected by MTT assay. Morphological features of apoptosis were analyzed, with Hoechst 33258/ Propidium iodide dual staining, The level of intracellular free calcium ([Ca2+]i) was calculated by Fura-2/AM fluorescence ratio imaging. Results: ① The viability of PC-12 cells was significantly decreased in proportion to concentration of Aβ25-35 and duration of exposure to Aβ25-35. ② The apoptotic cells appeared in a time and concentration-dependent manner, and the maximal apoptosis happened at 48 h after exposure to 20 μmol/L of Aβ25-35 and 36 h to 30 μmol/L, Cell death reached the peak at 12-24 h later than the apoptotic peak. (3) [Ca2+]i of PC-12 cells was increased in proportion to duration of exposure to the same concentration of Aβ25-35. The time of the highest increase rate of [Ca2+]i was about 12 h earlier than that of apoptosis. Conclusion: An AD cell model using the PC-12 cells induced with Aβ25-35 displays a series of changes related to apoptosis, which may be related to elevation of [Ca2+]i.

关键词： β-amyLoid protein intracellular free calcium apoptosis Alzheimer dis-ease cell model