1.Effects of conotoxin SO3 on free calcium ion concentration in rat cultured hippocampal neurons induced by hypoxia.
Shi-Wei JIANG ; Xiao-Wei ZHOU ; Ai-Shi DING
Chinese Journal of Applied Physiology 2005;21(4):375-422
Animals
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Calcium
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metabolism
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Cell Hypoxia
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Cells, Cultured
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Hippocampus
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cytology
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Neurons
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drug effects
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metabolism
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Rats
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Rats, Wistar
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omega-Conotoxins
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pharmacology
3.A method for culturing neonatal mice cardiomyocytes in vitro and its applications in toxicity evaluation
Hai-ying, YANG ; Wei, DING ; Ai-shi, DING ; Shuang-qing, PENG
Bulletin of The Academy of Military Medical Sciences 2010;34(1):30-33
Objective To establish a method for culturing neonatal mice cardiomyocytes, and construct an in vitro model of cardiomyocytes for assessing the cardiac toxicity of chemicals. Methods Hearts of neonatal mice of 1 day old were digested with enzyme mixture of trypsin/collagenase type Ⅱ/dispase and the cell suspensions were pre-plated to flask for a short time and then seeded on coated dishes. The cultured cells were treated with 5-fiuorine-deoxy-uridine(15 μg/ml)and uridine(35 μg/ml)to enrich cardiomyocytes that were identified according to the morphology and immunocytochemistry of α-actin antigen. Cardiac myocytes were incubated with 0-64 μg/ml of a fusarium mycotoxin butenolide(BUT) for 12 h. Cell viability was then evaluated by MTT assay. Microscopic observation showed that BUT induced significant morphological changes including cellular swelling, vacuolation and breakage of muscle fibers. Results It was found that 96% of the cultured cells were cardiomyocytes and the myocytes kept beating after 90 days of culture. Concentration-dependent decreases in cell viability following exposure of cardiac myocytes to BUT were observed. Conclusion The results indicated that relatively pure primary culture of neonatal mice cardiomyocytes is successfully established. BUT possesses the potential to induce myocardial toxicity.
4.Effects of Extracorporeal Shock Wave Therapy on Early to Mid-term Knee Osteoarthritis: A Randomized Controlled Clinical Tri-al
Zhe ZHAO ; Zhan SHI ; Jun YAN ; Quan AI ; Zekun DING ; Gengyan XING
Chinese Journal of Rehabilitation Theory and Practice 2014;20(1):76-78
Objective To investigate the effect of extracorporeal shock wave therapy (ESWT) on early and mid-term knee osteoarthritis (KOA). Methods 70 patients were randomly assigned to ESWT group (n=34) or control group (n=36). The ESWT group was given shock wave on unilateral knee, while the control group accepted shock wave with the energy density of 0. They were evaluated with Visual Analog Scale (VAS) on movement, Lequesne Index and Western Ontario and McMaster Universities (WOMAC) Osteoarthritis Index before and 12 weeks after the intervention. Results The scores of VAS, Lequesne Index and WOMAC Osteoarthritis Index improved more in the ESWT group than in the control group after intervention (P<0.01). Conclusion ESWT is effective on KOA as a non-surgical treatment.
5.Serum low-density lipoprotein and heart rate related to the effectiveness in the treatment of angina pectoris with two similar Chinese medicine herbal formulae.
Shi-yan YAN ; Chen YAO ; Xiao-rong DING ; Miao JIANG ; Ai-ping LU
Chinese journal of integrative medicine 2011;17(12):903-907
OBJECTIVETo explore the effectiveness related indicators which might help identify the indications of Tongxinluo Capsule () and Kangxin Capsule () targeting on qi deficiency and blood stasis pattern in Chinese medicine (CM) in the treatment of angina pectoris.
METHODSThe data from a multicenter, randomized and double-blinded study conducted at 5 centers in China were obtained for the analysis. A total of 239 patients with angina pectoris and CM syndrome of qi deficiency and blood stasis were randomly assigned in a 1:1 ratio to Tongxinluo Capsule group (119 cases) and Kangxin Capsule group (120 cases). Angina effectiveness and electrocardiogram (ECG) improvement were selected as the therapeutic outcomes.
RESULTSAfter a 4-week treatment, the effective rates of Tongxinluo Capsule and Kangxin Capsule were 43.70% and 25.00%, respectively (P <0.05). Serum low-density lipoprotein (LDL) level was found to influence the effectiveness of Tongxinluo Capsule which had higher effective rate in the patients with lower level of LDL. Heart rate was found to influence the effectiveness in the patients treated with Kangxin Capsule which had higher effective rate in the patients with heart rate [Symbol: see text]80 beats/min.
CONCLUSIONLDL level and heart rate were the indicators which help indentify the indications of Tongxinluo Capsule and Kangxin Capsule, respectively, in the treatment of angina pectoris with CM syndrome of qi deficiency and blood stasis.
Adult ; Aged ; Angina Pectoris ; blood ; drug therapy ; physiopathology ; Drugs, Chinese Herbal ; pharmacology ; therapeutic use ; Female ; Heart Rate ; drug effects ; physiology ; Humans ; Lipoproteins, LDL ; blood ; Male ; Middle Aged ; Patient Dropouts ; Platelet Count ; Treatment Outcome
6.CoCl2-induced enhancement of glucose transport activity in mediating hypoxic tolerance in cultured hippocampal neurons.
Shun YU ; Ming FAN ; Tong ZHAO ; Ai-Shi DING ; Fu-Zhuang WANG
Acta Physiologica Sinica 2002;54(6):508-512
The effect of CoCl(2) pretreatment on glucose transport activity of cultured newborn rat hippocampal neurons and its role in neuronal hypoxic tolerance were observed. The results showed that the 2-deoxy-D-[1-(3)H ]glucose uptake rate and the mRNA expressions of glucose transporters (GLUT1 and GLUT3) in the hippocampal neurons were significantly increased after a 24-hour pretreatment with CoCl(2). The cell injury induced by 6-hour or 8-hour hypoxic exposure was also greatly reduced by CoCl(2) pretreatment. The protective effect of CoCl(2) on the neurons was largely abolished by cytochalasin B, a specific inhibitor of glucose transporters. The results suggest that CoCl(2) can increase mRNA expressions of GLUT1 and GLUT3 and glucose transporter activity of the neurons, which may be an important mechanism for the increased tolerance of the neurons to hypoxia.
Animals
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Animals, Newborn
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Cell Hypoxia
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Cell Survival
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drug effects
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Cells, Cultured
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Cobalt
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pharmacology
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Glucose Transporter Type 1
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metabolism
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Glucose Transporter Type 3
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metabolism
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Hippocampus
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cytology
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Hypoxia
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metabolism
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Neurons
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drug effects
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metabolism
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Organometallic Compounds
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pharmacology
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RNA, Messenger
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genetics
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Rats
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Rats, Wistar
7.Effects of rhIL-6 on Bcl-2 and Bax expression and apoptosis after anoxia-reoxygenation in cultured rat hippocampal neurons.
Ai-Shi DING ; Fu-Zhuang WANG ; Li-Ying WU ; Ming FAN
Acta Physiologica Sinica 2002;54(2):115-120
The purpose of the present study was to determine the effects of recombinant human interleukin-6 (rhIL-6) on the Bcl-2 and Bax expression and apoptosis after anoxia-reoxygenation in cultured rat hippocampal neurons. The control and rhIL-6 treated hippocampal neurons cultured for 12 d were exposed to anoxia environment (90% N2+10% CO2) for 2 and 4 h and then were reoxygenated for 24 and 72 h. The expression of Bcl-2 and Bax was revealed immunocytochemically using the antiserum against Bcl-2 and Bax. The apoptosis was examined by the terminal deoxynucleotidyl transferase-mediated biotinylated deoxyuridine triphosphate nickel end labeling (TUNEL) method and flow cytometric analysis. The results showed that in cultured hippocampal neurons the Bcl-2 expression decreased while Bax expression and the percentage of apoptotic neurons increased after anoxia-reoxygenation compared with those before anoxia. In comparison with the control, after anoxia-reoxygenation the Bcl-2 expression in hippocampal neurons was higher than that in rhIL-6 group; however the Bax expression and the percentage of the apoptosis were decreased in rhIL-6 group. It is suggested that rhIL-6 may play a role in protecting neurons from the damage induced by anoxia-reoxygenation.
Animals
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Apoptosis
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drug effects
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Cell Hypoxia
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drug effects
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physiology
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Cells, Cultured
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Hippocampus
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cytology
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Interleukin-6
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pharmacology
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Neurons
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drug effects
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physiology
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Proto-Oncogene Proteins
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biosynthesis
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Proto-Oncogene Proteins c-bcl-2
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biosynthesis
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Rats
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Rats, Wistar
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Recombinant Proteins
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pharmacology
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bcl-2-Associated X Protein
8.Establishment of the model of oxygen-glucose deprivation in vitro rat hippocampal neurons.
Yan WU ; Ai-shi DING ; Li-ying WU ; Zi-min MA ; Ming FAN
Chinese Journal of Applied Physiology 2003;19(2):197-200
AIMTo establish the model of oxygen-glucose deprivation in vitro rat hippocampal neurons.
METHODSThe hippocampal neurons cultured for 12 d were exposed to combined oxygen-glucose deprivation for 0.5-4 h and then cultured with original medium in normoxia for 24 h. Auto-biochemical analyzer determined LDH activity. The change of neuronal morphology and neuron survival were observed by converted contrast microscope and assessed by photography analysis system. Neuron apoptosis was detected by using the terminal deoxynucleotidyl transferase-mediated biotinylated deoxyuridine triphosphate nickel end labeling (TUNEL) method.
RESULTSThe neurons swelled, LDH release increased and neuron survival decreased after gradually oxygen-glucose deprivation. The percentage of apoptosis increased obviously 24 h after recovering the supply of oxygen and glucose.
CONCLUSIONThe model of oxygen-glucose deprivation in vitro rat hippocampal neurons is established successfully by using the modified ACSF (artificial cerebral spinal fluid) with serum and glucose free.
Animals ; Animals, Newborn ; Cell Hypoxia ; Cells, Cultured ; Glucose ; deficiency ; Hippocampus ; cytology ; Neurons ; cytology ; Oxygen ; physiology ; Rats ; Rats, Wistar
9.Protective effect of Quinacrine on striatum neurons from heat treatment injury.
Yong-Qi ZHAO ; Yan WU ; Shu-Hong LIU ; Xue-Ming GE ; Ai-Shi DING ; Ming FAN
Chinese Journal of Applied Physiology 2004;20(4):319-323
AIMTo study the protective effect of Quinacrine(QA) on rat striatum neurons from the injury caused by heat environment treatment, to probe the relationship between cell membrane injury and cellular injury protection, and to seek the possibility of QA as a preventive agent to heat injury.
METHODSPrimary cultured striatum neurons from newborn rats were pretreated with QA at different concentration for 1 h, and then heat-treated at 43 degrees C for another 1 h. Cell necrosis was detected by Trypan blue staining, and apoptosis was evaluated through Activated Caspase-3 dye and TdT dye.
RESULTSHeat treatment effected the survival of striatum neurons and resulted in great number of cell death, which was mainly mediated by cell necrosis process. It was shown that treatment of QA itself had little effect on the survival of striatum neurons, while QA pretreatment decreased cellular necrosis caused by following heat treatment.
CONCLUSIONQA protects striatum neurons from heat environment injury at about 20 pmol/L, and the protection may mediated by reduction of necrosis.
Animals ; Apoptosis ; drug effects ; Caspase 3 ; metabolism ; Cell Death ; drug effects ; Cells, Cultured ; Corpus Striatum ; cytology ; Heat-Shock Response ; Neurons ; drug effects ; Quinacrine ; pharmacology ; Rats ; Rats, Wistar
10.Effects of hypoxia on the proliferation of embryonic stem cells.
Hui-qing ZHAO ; Ling-ling ZHU ; Tong ZHAO ; Li-ying WU ; Ai-shi DING ; Ming FAN
Chinese Journal of Applied Physiology 2004;20(3):209-213
AIMTo investigate the effects of hypoxia on the proliferation of mouse embryonic stem cells (mouse ES cells) in vitro.
METHODSWe observed the proliferation of ES cells by hematometery and BrdU-labeled flow cytometry (FCM), and we also detected the expression of hypoxia inducible factor-1a (HIF-1a) by RT-PCR.
RESULTS(1) The number of ES cells after culturing in the hypoxia environment (3% O2 and 10% O2) for 24 hours were lesser than those in normoxia (20% O2). (2) The number of ES cells significantly increased after intermittent hypoxia (3% O2) stimulus for 10 minutes per day for 4 days. (3) We also observed the relation between the expression of HIF-1a and the proliferation of ES cells by RT-PCR. The results showed that the expression of HIF-1a had no significant change after ES cells were culturing in hypoxia environment (3% O2 and 10% O2) for 24 hours or in intermittent hypoxia (3% O2 and 10% O2) for 4 days.
CONCLUSIONThese results suggest that intermittent hypoxia (3% O2) can significantly promote the proliferation of ES cells in vitro, while persistent hypoxia inhibits those, and the mechanism of these should be addressed in further.
Animals ; Cell Hypoxia ; Cell Proliferation ; Cells, Cultured ; Embryonic Stem Cells ; cytology ; Mice