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5-Hydroxydecanoate inhibits proliferation of hypoxic human pulmonary artery smooth muscle cells by blocking mitochondrial KATP channels

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Acta Pharmacologica Sinica 2007年第10期28卷 1531-1540页

作者： Tao WANG~2,Zhen-xiang ZHANG~(2,4),Yong-jian XU~2,Qing-hua HU~3~2 department of Respiratory Medicine,Tongji Hospital,Tongji Medical college,huazhong university of science and technology,wuhan430030,china ~3 key laboratory of pulmonary diseases of the ministry of health of china and department of pathophysiology,tongjimedical college,huazhong university of science and technology wuhan 430030,china department of Respiratory Medicine Tongji Hospital Tongji Medical College Huazhong University of Science and Technology Wuhan China key laboratory of pulmonary diseases of the ministry of health of china and department of pathophysiology Tongji Medical College Huazhong University of Science and Technology Wuhan China

Aim:To study the effect of 5-hydroxydecanoate （5-HD） on the proliferation of 24h hypoxic human pulmonary artery smooth muscle cells （HPASMC） and to explorethe pharmacological mechanisms of 5-HD as an inhibitor of mitochondrial mem-brane ATP-sensitive potassium channel ***:Normoxic or hy-poxic HPASMC in culture were stimulated by either diazoxide or 5-HD for 24 *** proliferation of HPASMC was examined by 3-（4,5-dimethyl-2-thiazol-yl）-2,5-diphenyl-2H-tetrazolium bromide （MTT） assay and proliferating cell nuclear anti-gen （PCNA） immunohistochemistry *** apoptosis of HPASMC wasassessed by terminal deoxynucleotidyl transferase-mediated dUTP nick end la-beling （TUNEL） assayand flow cytometric *** relative changes inmitochondrial membrane potential （ΔΨm） were measured using the rhodamine fluo-rescence （R-123） ***:Both hypoxia and diazoxide stimulationincreased ΔΨmvalue measured by the absorbance of MTT,PCNA-positive stain-ing and decreased TUNEL-positive staining and apoptotic cells in *** and the concomitant stimulation of diazoxide obviously enhanced theeffects of hypoxia or diazoxide alone.5-HD significantly attenuated the effects ineach of the above ***,5-HD partially inhibited the effect ofhypoxia on R-123 fluorescence intensity in ***:5-HD caninhibit the proliferation of hypoxic HPASMC by blocking mitochondrial KATPchannels.

关键词： pulmonary artery smooth muscle cell hypoxia 5-hydroxydecanoate mitochondrial membrane ATP-sensitive potassium channel

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Passive sensitization increases histamine-stimulated calcium signaling and NF-κB transcription activity in bronchial epithelial cells

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Acta Pharmacologica Sinica 2006年第6期27卷 708-714页

作者： Si JIN~(2,5)Dan TIAN~3 Jian-guo CHEN~4 Li-ping ZHU~2 Sheng-yuan LIU~2 Di-xun WANG~2 2 department of pathophysiology,Tongji Medical college,huazhong university of science and technology key laboratory of pulmonary Disease of ministry of health of china,wuhan 430030,china 3 department of Pathology,Tongji Medical college.huazhong university of science and technology.wuhan 430030,china 4 department of Pharmacology,Tongji Medical college,huazhong university of science and technology,wuhan 430030,china department of pathophysiology Tongji Medical College Huazhong University of Science and Technology key laboratory of pulmonary Disease of ministry of health of china Wuhan China department of Pathology Tongji Medical College Huazhong University of Science and Technology Wuhan China department of Pharmacology Tongji Medical College Huazhong University of Science and Technology Wuhan China

Aim:To find out if the two aspects of asthma（chronic airway inflammation andbronchial hyperresponsiveness）are related to hypersensitivity of calcium signal-ing in bronchial epithelial ***:Porcine bronchial epithelial cells（PBEC）were divided into sensitized（S）and nonsesitized（N）*** group S,the cellswere preincubated with serum from ovalbumin sensitized guinea *** group N,the cells were preincubated with serum from nonsensitized guinea *** calcium imaging and ELISA-based NF-κ3 activity were used to evaluate thehistamine-stimulated intracellular free calcium level and NF-κB activity,***:First,0.1μmol/L histamine could induce [Ca2+]ioscillationsin PBEC of group S,but not in group ***,1/Jmol/L histamine could induce[Ca2+]ioscillations of PBEC in both group S and group *** [Ca2+]ioscillationfrequency of PBEC was significantly higher in group S than in group N,thoughthe[Ca2+]ioscillation amplitude showed no difference between the two ***,when 10 μmol/L histamine was used to stimulate PBEC a transient initialincrease followed by a sustained elevation（FSE）of [Ca2+]iwas observed in PBECin both *** amplitude of the FSE of [Ca2+]iin PBEC was significantlyhigher in group S than in group *** subsequent NF-κB activity was in accor-dance to the calcium oscillation frequency evoked by histamine,but not to ***:It was suggested that the increased sensitivity of calciumsignaling in bronchial epithelial cells might contribute to the exorbitant inflamma-tion or increased susceptibility in asthmatic airway epithelial cells.

关键词： asthma histamine calcium signaling bronchi epithelial cell nuclear factor of kappaB

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Role of melatonin in Alzheimer-like neurodegeneration

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Acta Pharmacologica Sinica 2006年第1期27卷 41-49页

作者： Jian-zhi WANG~(2,4) Ze-fen WANG~(2,3)2 pathophysiology department,key laboratory of Neurological diseases of Hubei Province,Tongji Medical School,huazhong university of science and technology,wuhan 430030,china 3 department of Physiology,Medical college,wuhan university wuhan 430071,china pathophysiology department Key Laboratory of Neurological Diseases of Hubei Province Tongji Medical School Huazhong University of Science and Technology Wuhan China department of Physiology Medical College Wuhan University Wuhan China

Alzheimer disease (AD),an age-related neurodegenerative disorder with progres-sive loss of memory and deterioration of comprehensive cognition,is character-ized by extracellular senile plaques of aggregated β-amyloid (Aβ),and intracellularneurofibrillary tangles that contain hyperphosphorylated tau *** showed that melatonin,an indoleamine secreted by the pineal gland,mayplay an important role in aging and AD as an antioxidant and *** decreases during aging and patients with AD have a more profoundreduction in this *** from clinical trials indicate that melatonin supple-mentation improves sleep,ameliorates sundowning,and slows down the progres-sion of cognitive impairment in Alzheimer's *** efficiently pro-tects neuronal cells from Aβ-mediated toxicity via antioxidant and anti-amyloidproperties:it not only inhibits Aβ generation,but also arrests the formation ofamyloid fibrils by a structure-dependent interaction with Aβ.Our recent studieshave demonstrated that melatonin efficiently attenuates Alzheimer-like *** the exact mechanism is still not fully understood,a direct regulatory influence of melatonin on the activities of protein kinases andprotein phosphatases is ***,melatonin also plays a role inprotecting cholinergic neurons and in ***,the neuroprotectiveeffects of melatonin and the underlying mechanisms by which it exerts its effectsare *** capacity of melatonin to prevent or ameliorate tau and Aβpathology further enhances its potential in the prevention or treatment of AD.

关键词： Alzheimer disease melatonin tau,beta amyloid

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A template of rat brain based on fMRI T2* imaging

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Progress in Natural science:Materials International 2005年第6期15卷 502-506页

作者： HU Zhenghui1, WU Yigen2, WANG Xiaochuan3, WANG Jianzhi3, CHEN Feiyan1 and TANG Xiaowei1*(1. department of Physics and Bio-X laboratory, Zhejiang university, Hangzhou 310027, china 2. Institute of High Energy Physics and key laboratory of Nuclear Analysis Techniques, Chinese Academy of sciences,Beijing 100039, china 3. department of pathophysiology, Tongji Medical college, huazhong university of science and technology, wuhan 430030, china) department of Physics and Bio-X laboratory Zhejiang University Hangzhou 310027 China key laboratory of Nuclear Analysis Techniques Institute of High Energy Physics Chinese Academy of Sciences Beijing 100039 China department of pathophysiology Tongji Medical College Huazhong University of Science and Technology Wuhan 430030 China

The development of functional magnetic resonance imaging (fMRI) technology has made it possible to carry out functional brain imaging experiments in small animals. Usually, group data is required to form the assessment of population, which can not only increase the sensitivity of the overall experiment, but also allow the generalization of the conclusion to the whole population. In order to average the signals of functional brain images from different subjects, it is necessary to put all the mapping images into the same standard space (template image). However, up to now, most animal brain templates remain unavailable and it must be done by ourselves. In this study, a template image based on the brains of eight male Wistar rats is obtained, and it is successfully used in our present Alzheimer disease (AD)-like rat model studies as template for spatially normalizing images to the same stereotaxical space. The fMRI results processed with statistical parametric mapping (SPM99) software are in agreement with the results from immunohistochemical experiment, which proves that this method is universally applicable to the pathologic models of other small animals and to human brain lesion studies.

关键词： fMRI animal model atlas template normalization T*2.

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Correlation of Behavior Changes and BOLD Signal in Alzheimer-like Rat Model

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Acta Biochimica et Biophysica Sinica 2004年第12期36卷 803-810页

作者： Zheng-Hui HU#, Xiao-Chuan WANG1#, Li-Yun LI3, Mai-Li LIU3, Rong LIU1, Zhiqun LING1, Qing TIAN1, Xiao-Wei TANG, Yi-Gen WU2*, and Jian-Zhi WANG1* department of Physics and Bio-X disciplinary laboratory, Zhejiang university, Hangzhou 310027,china 1 department of pathophysiology, Tongji Medical college, huazhong university of science and technology, wuhan 430030, china 2Institute of High Energy Physics and key laboratory of Nuclear Analysis Techniques, Chinese Academy of sciences, P.O. Box 918, Beijing 100039, china 3wuhan Institute of Physics and Mathematics, State key laboratory of Magnetic Resonance and Atomic and Molecular Physics, Chinese Academy of sciences, wuhan 430071, china department of Physics and Bio-X disciplinary laboratory Zhejiang UniversityHangzhou 310027China department of pathophysiology Tongji Medical College Huazhong University of Science and TechnologyWuhan 430030 China wuhan Institute of Physics and Mathematics State Key Laboratory of Magnetic Resonance and Atomic and Molecular Physics Chinese Academy of SciencesWuhan 430071 China Institute of High Energy Physics and key laboratory of Nuclear Analysis Techniques Chinese Academy of SciencesP.O. Box 918 Beijing 100039 China

To explore a potential means for the early diagnosis of Alzheimer disease, we studied the relationship of resting T2* signal and tau hyperphosphorylation/spatial memory deficit. The rat model with tau hyperphosphorylation and spatial memory deficit was established by bilateral hippocampi injection of isoproterenol (IP). Then, the correlative alteration between resting T2* signal and spatial memory retention was assessed with blood oxygenation level dependent (BOLD) functional magnetic resonance imaging (fMRI) study and Morris Water Maze test, and Western blot was employed to confirm tau hyperphosphorylation. The analysis showed following results. (1) Tau phosphorylation at Ser396/Ser404 and Ser199/Ser202 was signifi- cantly increased in IP-injected rats as detected by PHF-1 and tau-1, respectively. (2) An AD-like spatial memory retention disturbance was induced at 24 h after isoproterenol injection. (3) A sensitivity threshold of resting T2* signal intensity, which separated the IP-treated rats from vehicle control, was obtained by applying linear regression analysis, and an estimated sensitivity statistical threshold was at 32.62. These results suggest that resting T2* signal may serve as a noninvasive quantitative marker in predicting AD-like spatial memory deficits and tau hyperphosphorylation.

关键词： Alzheimer disease (AD) functional magnetic resonance imaging (fMRI) isoproterenol (IP) Morris Water Maze diagnosis

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5-Hydroxydecanoate inhibits proliferation of hypoxic human pulmonary artery smooth muscle cells by blocking mitochondrial KATP channels

5-Hydroxydecanoate inhibits proliferation of hypoxic human p...

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湖北省暨武汉市病理生理学会第十四届学术年会

作者： Tao WANG~2,Zhen-xiang ZHANG~(2,4),Yong-jian XU~2,Qing-hua HU~3 2 department of Respiratory Medicine,Tongji Hospital,Tongji Medical college,huazhong university of science and technology,wuhan 430030,china 3 key laboratory of pulmonary diseases of the ministry of health of china and department of pathophysiology,Tongji Medical college,huazhong university of science and technology,wuhan 430030,china

Aim:To study the effect of 5-hydroxydecanoate(5-HD)on the proliferation of 24 h hypoxic human pulmonary artery smooth muscle cells(HPASMC)and to explore the pharmacological mechanisms of 5-HD as an inhibitor of mitochondrial membrane ATP-sensitive potassium channel activation. Methods:Normoxic or hypoxic HPASMC in culture were stimulated by either diazoxide or 5-HD for 24 *** proliferation of HPASMC was examined by 3-(4,5-dimethyl-2-thiazol-yl)-2,5-diphenyl-2H -tetrazolium bromide(MTT)assay and proliferating cell nuclear antigen(PCNA) immunohistochemistry *** apoptosis of HPASMC was assessed by terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling(TUNEL)assay and flow cytometric *** relative changes in mitochondrial membrane potential(△Ψm)were measured using the rhodamine fluorescence(R-123)***:Both hypoxia and diazoxide stimulation increased△Ψm value measured by the absorbance of MTT,PCNA-positive staining and decreased TUNEL-positive staining and apoptotic cells in *** and the concomitant stimulation of diazoxide obviously enhanced the effects of hypoxia or diazoxide alone.5-HD significantly attenuated the effects in each of the above ***,5-HD partially inhibited the effect of hypoxia on R-123 fluorescence intensity in ***:5-HD can inhibit the proliferation of hypoxic HPASMC by blocking mitochondrial Kchannels.

关键词： pulmonary artery smooth muscle cell hypoxia 5-hydroxydecanoate mitochond-rial membrane ATP-sensitive potassium channel

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VEGF stimulates calcium oscillations and activates NFAT in human pulmonary valve endothelial Cells

VEGF stimulates calcium oscillations and activates NFAT in h...

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第八届海峡两岸心血管科学研讨会

作者： Lei CAI,Li-ping ZHU,Qing-hua HU~# (department of pathophysiology,key laboratory of pulmonary diseases of ministry of health of china and department of pathophysiology,Tongji Medical college,huazhong university of science & technology,wuhan 430030,china)

Calcific aortic stenosis(AS),previously thought of as a degenerative disease,results from an active process involving cell proliferation and the expression of an osteoblast phenotype in diseased valve leaflets. The central roles of neoangiogenesis and bone formation in the pathobiology of AS suggest the involvement of vascular endothelial growth factor(VEGF),known to regulate endothelial cell proliferation and the expression of bone - forming *** hypothesized that VEGF leads to cell proliferation in cardiac valves by stimulating the Ca- dependent activation of the transcription factor NFAT which is involved in VEGF - induced proliferation of valvular endothelial *** also hypothesized that this would be inhibited by an HMG CoA reductase inhibitor(i.e.a"statin"),since statins inhibit proliferative changes in a rabbit model of AS and decrease progression of AS in ***(100 ng/mL) - stimulated human pulmonary valve endothelial cells(HPVEC ) loaded with the Ca - sensitive fluorescent indicator fura - 2 demonstrated repetitive intracellular Ca oscillations that were blocked by an overnight exposure to 10?? M *** also stimulated the cytoplasmic - to - nuclear translocation of a GFP - NFAT fusion construct in HPVEC,which was inhibited by *** reversed the inhibitory effects of *** data suggest that VEGF - stimulated valvular endothelial cell proliferation and NFAT activation may play a role in the pathobiology of AS and point to a mechanism whereby statins may inhibit this process.

关键词： calcific aortic stenosis vascular endothelial growth factor human pulmonary valve endothelial cells

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Dracorhodin Perchlorate Suppresses Proliferation and Induces Apoptosis in Human Prostate Cancer Cell Line PC-3

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Journal of huazhong university of science and technology(Medical sciences) 2011年第2期31卷 215-219页

作者：何远桥鞠文郝华刘庆吕磊曾甫清 department of laboratory Animal science Nanchang University department of Urology Union Hospital Tongji Medical College Huazhong University of Science and Technology department of pathophysiology Tongji Medical College Huazhong University of Science and Technology

The growth inhibition and pro-apoptosis effects of dracorhodin perchlorate on human prostate cancer PC-3 cell line were examined. After administration of 10-80 μmol/L dracorhodin perchlorate for 12-48 h, cell viability of PC-3 cells was measured by MTT colorimetry. Cell proliferation ability was detected by colony formation assay. Cellular apoptosis was inspected by acridine orange-ethidium bromide fluorescent staining, Hoechst 33258 fluorescent staining, and flow cytometry （FCM） with annexin Ⅴ-FITC/propidium iodide dual staining. The results showed that dracorhodin perchlorate inhibited the growth of PC-3 in a dose- and time-dependent manner. IC50 of dracorhodin perchlorate on PC-3 cells at 24 h was 40.18 μmol/L. Cell clone formation rate was decreased by 86% after treatment with 20 μmol/L of dracorhodin perchlorate. Some cells presented the characteristic apoptotic changes. The cellular apoptotic rates induced by 10-40 μmol/L dracorhodin perchlorate for 24 h were 8.43% to 47.71% respectively. It was concluded that dracorhodin perchlorate significantly inhibited the growth of PC-3 cells by suppressing proliferation and inducing apoptosis of the cells.

关键词： dracorhodin perchlorate prostate cancer proliferation apoptosis

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Inhibitory Activity of Nuclear Factor-κB Potentiates Cisplatin-induced Apoptosis in A549 Cells

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Journal of huazhong university of science and technology(Medical sciences) 2008年第3期28卷 251-256页

作者：张建徐永健熊维宁张珍祥杜春玲乔礼芬倪望陈士新 department of Respiratory Medicine Tongji HospitalTongji Medical CollegeHuazhong University of Science and TechnologyKey Laboratory of Pulmonary Diseases of Ministry of Health of China

Whether inhibiting the activity of nuclear factor （NF）-κB potentiates cisplatin-induced apoptosis in non-small cell lung cell line A549 cells was investigated. The recombinant plasmid pcDNA3.1（＋）/IκBα expressing IκBα was constructed. The in vitro cultured A549 cells were transfected with pcDNA3.1 （＋）/IκBα alone, or pcDNA3.1（＋）/IκBα combined with cisplatin. The mitochondrial membrane potential （△ψm） was determined by rhodamine 123, the activity of caspase-3 was tested by colorimetric assay, and cell apoptosis was detected by flow cytometry with the annexin V/propidium iodide assay. The results showed that the activity of NF-κB in A549 cells was inhibited by transfecting pcDNA3.1（＋）/IκBα. Transfection of pcDNA3.1（＋）/IκBα alone did not promote apoptosis. Treatment of cisplatin alone had a little effect on cell apoptosis. Transfection of pcDNA3.1（＋）/IκBα combined with cisplatin treatment significantly induced apoptosis of A549 ceils. It was concluded that inhibiting the activity of NF-κB potentiated cisplatin-induced apoptosis of A549 cells.

关键词： nuclear factor-kappaB lung neoplasms cisplatin apoptosis

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Extracellular Calcium-sensing Receptor is Critical in Hypoxic pulmonary Vasoconstriction

Extracellular Calcium-sensing Receptor is Critical in Hypoxi...

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第八届海峡两岸心血管科学研讨会

作者： Ji-wei Zhang~△,Juan Zhou~△,Yan-kai Lu,Tao Wang,Li-ping Zhu,Qing-hua Hu~# (department of pathophysiology,key laboratory of pulmonary diseases of ministry of health and key laboratory of Environment and health of ministry of Education, Tongji Medical college,huazhong university of science and technology,wuhan 430030,china)

The initiation of hypoxic pulmonary vasoconstriction relies mainly on an increase in cytosolic calcium ([Ca]) in pulmonary artery smooth muscle *** of the processes completely depend on extracellular *** Ca can be sensed by extracellular calcium - sensing *** potential role of extrtacelllular Ca sensing pathway in hypoxic pulmonary vasoconstriction has never been *** study aims to determine whether extracellular calcium - sensing receptor is pivotal in the initiation of hypoxic pulmonary vasoconstriction. Experiments were performed in cultured pulmonary artery smooth muscle cells,isolated pulmonary arteries and rats including extracellular calcium - sensing receptor knockdown *** hypoxia and HO equivalent to the level achieved by hypoxia increased[Ca] in an extracellular Ca- dependent manner in pulmonary artery smooth muscle cells and this was inhibited by extracellular calcium - sensing receptor knockdown or its negative allosteric modulator,*** - increased HO generation was diminished by mitochondria *** depletion abolished hypoxia - induced[Ca] increase, which was reversed by HO *** calcium - sensing receptor knockdown or Calhex231 however prevented the reversible effect of *** - induced[Ca] increase was abolished by PEG-Catalase, not PEG-SOD pretreatment,attenuated by ryanodine receptor3 - knockdown or inhibition of store -operated Ca *** pulmonary vasoconstriction in vitro and in vivo was inhibited by Calhex 231 and by extracellular calcium - sensing receptor knockdown. In conclusion,the activation of mitochondrial HO sensitized extracellular calcium - sensing receptor by extracellular Ca mediates hypoxia - induced[Ca] increase in pulmonary artery smooth muscle cells and thus initiates hypoxic pulmonary vasoconstriction.

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