1.Inhibitory effect of taurine in hypoxia-induced rat pulmonary artery smooth muscle cell proliferation and signal transduction mechanism.
Xiao-Dan ZHANG ; Peng SUN ; Da-Ling ZHU ; Nan XIE
China Journal of Chinese Materia Medica 2014;39(10):1902-1907
OBJECTIVETo discuss the effect of taurine (Tau) on the proliferation of hypoxia-induced pulmonary artery smooth muscle cells (PASMCs), and study whether the extracellular signal-regulated kinase 1/2 (ERK1/2) signal pathway participated in the Tau-inhibited PASMC proliferation process and the possible molecular mechanism.
METHODThe primary culture was performed for PASMCs in rats. The second to fifth generations were adopted for the experiment. The Tau concentration was 80 mmol x L(-1). The concentration of ERK1/2 blocker (PD98059) was 50 micromol x L(-1). The drug administration time was 24 h. The effect of Tau on the PASMC proliferation was detected by MTT assay, immunofluorescence staining method and western blot under different conditions. The PASMCs were growing were divided into four groups: the normoxia group, the normoxia + Tau group, the hypoxia group and the hypoxia + Tau group. The Western blot was adopted to detect whether the ERK1/2 signal pathway participated in the Tau-inhibited PASMC proliferation process. Subsequently, the PASMCs were divided into five groups: the normoxia group, the hypoxia group, the hypoxia + Tau group, the hypoxia + Tau + PD98059 group and the hypoxia + PD98059 group.
RESULTHypoxia could induce the PASMC proliferation. Under the conditions of normoxia, Tau had no effect on the PASMC proliferation. Under the conditions of normoxia and hypoxia, Tau had no effect on the expression of the tumor necrosis factor-alpha (TNF-alpha) among PASMCs. Tau could reverse the expression up-regulation of hypoxia-induced proliferative cell nuclear antigen (PCNA) (P < 0.01) and Cyclin A (Cyclin A) (P < 0. 05). Under the conditions of normoxia, Tau had no effect on the expression of phosphoryl extracellular signal-regulated kinase 1/2 (p-ERK1/2). Hypoxia could up-regulate the p-ERK1/2 expression (P < 0.01). Tau could reverse the up-regulation of the hypoxia-induced p-ERK1/2 expression(P < 0.01). Both PD98059 and Tau could inhibit the up-regulated expressions of PCNA, Cyclin A and p-ERK1/2. According to the comparison between the single addition of Tau and PD98059 under conditions of hypoxia, the hypoxia + Tau + PD98059 group showed more significant down-regulation in the expressions of PCNA, Cyclin A and p-ERK1/2.
CONCLUSIONTau could inhibit the hypoxia-induced PASMC proliferation, and may regulate it through ERK1/2 pathway.
Animals ; Cell Hypoxia ; drug effects ; Cell Proliferation ; drug effects ; Cells, Cultured ; MAP Kinase Signaling System ; drug effects ; Myocytes, Smooth Muscle ; cytology ; drug effects ; metabolism ; Oxygen ; metabolism ; Pulmonary Artery ; cytology ; drug effects ; metabolism ; physiopathology ; Rats ; Rats, Wistar ; Taurine ; pharmacology ; Tumor Necrosis Factor-alpha ; genetics ; metabolism
2.Expression of Heat-Shock Protein 70 in Kidney of Acute Injury Rats Caused by Gentamicin
hong-yan, ZHANG ; ling-di, SUN ; da-ya, ZHAO
Journal of Applied Clinical Pediatrics 1992;0(05):-
Objective To illustrate the expressional distribution of heat shock protein 70(HSP 70) in the kidney of acute renal injury rats caused by gentamicin.Methods One hundred and fifty rats were randomly divided into 3 groups: group A (normal group), group B [80 mg/(kg?d) gentamicin to be injected], group C [160 mg/(kg?d) gentamicin to be given]. The variations of renal pathology were observed at different time phasess by light scope and electromicroscope. Simultaneously, the expression of HSP 70 in kidney was evaluated by immunohistochemistry.Results HSP 70 distributed in epithelial cells of renal tubular in acute injury rats. The expression of HSP 70 increased markedly from the 6th hour after injection, peaked at the 12th hour and lasted for 48 hours. The expression of HSP 70 in group C was higher than that in group B(P
3.The Assembly of a Novel Enzyme Biosensor for Aflatoxin B_1 Detection
Da-Ling LIU ; Yi SHEN ; Jing ZHANG ; Dong-Sheng YAO ;
China Biotechnology 2006;0(03):-
A novel biosensor for aflatoxin B1 detecting has been reported. The biosensor electrode for AFB1 detecting was assembled by immobilized aflatoxin-oxidoreductase using open-ended multi-walled carbon nanotubes as matrix. Its linear range was between 0.16?M and 3.2?M. And if the specific anti-aflatoxin B1 antibody and aflatoxin oxidoreductase were both immobilized on the electrode with Multi-Walled carbon nanotubes, the detection limit of the modified electrode could be 16 nM with a 10 times improved sensitivity. The aflatoxin enzyme biosensor assembled this way strode one step forward its practical application.
4.Role of dynophin and substance P in end stage renal disease rats with pruritus
Xiaomei HUANG ; Ying ZHANG ; Xiaoyong ZHOU ; Da HE ; Shuihuai GUO ; Ling ZHANG ; Nijun ZHENG ; Lanqian XIE
Journal of Chinese Physician 2008;10(7):895-897
Objective To investigate the role of dynophin and substance P in pruritus with 5/6 subtotal nephrectomy rats (STNx). Methods 5/6 subtotal nephrectomy rots were prepared. After 24 weeks , STNx rats entered in end stage renal disease(ESRD). Sixty male SD rats were divided into 4 groups, STNx+P group(n=15), STNx+dynophin group( n=15), STNx control group(n=15) and sham operation group(n=15). Substance P (SP), dynophin and saline were intradermal injected separately. After injection, scratch reaction of rats in 30 minutes were recorded. And then SP concentration in blood was estimated by ELISA and SP expression in skin was examined by immunohistochemisty. Results SP concentration in blood of STNx rats were (1010.2±103.5)pg/ml, which increased to (2530.0± 236.3) pg/ml in STNx+SP group and decreased to (612.4±72.2)pg/ml in STNx+dynophin group, and it was the lowest in sham opera- tion group (240.2±36.5)pg/ml. SP expression in skin was similar to that of in blood. The scratch times in STNx+SP group were highest (7.3±1.9 times), there was no significant difference between STNx + dynophin group and STNx control group. Conclusion Pruritus in ESRD rats was correlated to the increase of SP in blood and skin. Intradermal injection of dynophin can decrease SP in blood and skin, but can not induce scratch in STNx rats.
5.Correlation study between ~(18)F-FDG uptake and hypoxia inducible factor-1α level,microvessel density in human gliomas
Li, CAI ; Chuan, ZHANG ; Yan-sheng, LI ; Da-cheng, LI ; Xi-ling, XING ; Shuo, GAO
Chinese Journal of Nuclear Medicine 2010;30(1):10-14
Objective To investigate the correlation between ~(18)F-fluorodeoxyglucose (FDG) uptake and hypoxia inducible factor1α (HIF-1α) level,microvessel density (MVD) in human gliomas.Methods ~(18)F-FDG PET scan was performed preoperatively in 41 patients with gliomas (including 23 highgrade and 18 low-grade tumors).The ratios of maximum standardized uptake value(SUV_(max))between tumor (T)and contralateral white matter (WM) were calculated (T/WM).Immunohistochemical stain methods were used to evaluate the level of HIF-1α and measure the MVD in tumors.Correlation analysis between SUV_(max) of T/WM and HIF-1α level,MVD wag performed.The t-test,one-way ANOVA test,Spearman rank correlation and Wilcoxon signed-rank test were calculated using SPSS 11.5 software.Results (1)The SUV_(max) of T/WM,HIF-1α level and MVD in high-grade and low-grade tumors groups were 3.39±1.43,95.7% and 44.13±16.1 vs 1.46±0.55.55.6% and 18.83±7.07,respectively.The difierences of SUV_(max) of T/WM,HIF-1α level and MVD between two groups were statistically significant (t=-5.921,z=-3.938,t=-6.745,all P<0.05).(2)Among 41 gliomas,the strong positive expression of HIF-1α was observed in 8,mederate in 9,weak in 15 and negative expression was found in 9,SUV_(max) of T/WM and MVD increased with increasing HIF-1α level.The differences of SUV_(max) of T/WM and MVD among 4 different groups were statistically significant (F=7.41,P<0.05).(3) The MVD of all gliomas was ranged from 9.76 to 94.52,which correlated with SUV_(max) of T/WM(r=0.759,P<0.05).Conclusions The SUV_(max) of T/WM correlates with HIF-1α level and MVD in gliomas.Therefore,~(18)F-FDG PET provides preoperatively a noninvasive assessment of hypoxia or angiogenesis in human glionma.
6.The Assembly of a Novel Enzyme Biosensor for Aflatoxin B1 Detection
Da-Ling LIU ; Yi SHEN ; Jing ZHANG ; Dong-Sheng YAO
China Biotechnology 2008;28(3):44-52
A novel biosensor for aflatoxin B1 detecting has been reported. The biosensor electrode for AFB1 detecting was assembled by immobilized aflatoxin-oxidoreductase using open-ended multi-walled carbon nanotubes as matrix. Its linear range was between 0.16μM and 3.2μM. And if the specific anti-aflatoxin B1 antibody and aflatoxin oxidoreductase were both immobilized on the electrode with Multi-Walled carbon nanotubes, the detection limit of the modified electrode could be 16 nM with a 10 times improved sensitivity. The aflatoxin enzyme biosensor assembled this way strode one step forward its practical application.
7.Effects of acupuncture at different acupoints on behaviors in depression model rats.
Jian-bin ZHANG ; Ling-ling WANG ; Mei LÜ ; Lan-ying LIU ; Da LI
Chinese Acupuncture & Moxibustion 2005;25(9):639-643
OBJECTIVETo observe therapeutic effects of acupuncture at different acupoints on depression.
METHODSSeventy adult male rats with similar Open-Field score were randomly divided into a normal control group, a model group, a blank control group, a drug treatment group, an acupuncture group I ["Baihui" (GV 20), "Shenting" (GV 24)], II ]"Neiguan" (PC 6), "Sanyinjiao" (SP 6)], and III ("Baihui", "Shenting", "Neiguan" and 'Sanyinjiao"), 10 rats in each group. Separation feeding, long term unpredictability and medium stimulation stress techniques were applied to develop depression model rats. Changes of behaviors were investigated with Open-Field method and sucrose consumption trial.
RESULTSCompared with the normal control group, the score for the behavior and the consumption of sucrose reduced in the depression model group; compared with the blank control group, the score for the behavior and the consumption of sucrose increased in all the treatment groups during treatment. There was no significant difference among the acupuncture groups in the increase of behavior scores 21 days after treatment (P > 0.05), and there were significant differences in the increase of consumption of sucrose in the acupuncture group I and the increase of score for vertical movement in the acupuncture group III 14 days after treatment (P < 0.05, P < 0.01).
CONCLUSIONAcupuncture can change the behavior abnormality induced by separation feeding, long term unpredictability and medium degree stimulation depression model rats and different acupoint selection is a factor in acuounture treatment.
Acupuncture Points ; Acupuncture Therapy ; Animals ; Depression ; therapy ; Depressive Disorder ; Humans ; Problem Behavior ; Rats
8.Kv3.4 channel is involved in rat pulmonary vasoconstriction induced by 15-hydroxyeicosatetraenoic acid.
Qian LI ; Hai-Rong BI ; Rong ZHANG ; Da-Ling ZHU
Acta Physiologica Sinica 2006;58(1):77-82
We have reported that hypoxia increases the activation of 15-lipoxygenase (15-LO), which converts arachidonic acid (AA) into 15-hydroxyeicosatetraenoic acid (15-HETE) in small pulmonary arteries (PAs). Through inhibition of Kv channels, 15-HETE causes more robust concentration-dependent contraction of PA rings from the hypoxic compared to the normoxic controls. However, the subtypes of Kv channels inhibited by 15-HETE are incompletely understood. The aim of the present study was to identify the contribution of Kv3.4 channel in the process of pulmonary vasoconstriction induced by 15-HETE using the tension studies of PA rings from rat with Kv3.4 channel blocker in tissue bath; to explore the role of vascular endothelium in15-HETE-induced pulmonary vasoconstriction through denuded endothelia of PA rings; and to define the downregulation of 15-HETE on the expression of Kv3.4 channel in cultured pulmonary artery smooth muscle cells (PASMCs) with RT-PCR and Western blot. In the present study, healthy Wistar rats were divided randomly into two groups: Group A with normal oxygen supply and group B with hypoxia. Six days later, the rats were killed. Pulmonary artery rings were prepared for organ bath experiments. Firstly, different concentrations of 15-HETE (10~1 000 nmol/L) were added to the Krebs solution. The isometric tension was recorded using a four-channel force-displacement transducer. Then Kv3.4 channel blocker, 100 nmol/L BDS-I, was added, followed by adding 1 mumol/L 15-HETE, and the isometric tension was recorded. Furthermore, RT-PCR and Western blot were employed to identify the influence of 15-HETE on the expression of Kv3.4 channel in cultured rat PASMCs.The results showed the PA tension was significantly increased both in groups A and B by 15-HETE in a concentration-dependent manner (P<0.05), especially in group B (P<0.05 compared to control); denuded endothelia enhanced 15-HETE concentration-related constrictions in rat PA rings; Kv3.4 channel blocker, BDS-I, significantly decreased the PA ring constriction induced by 15-HETE (P<0.05); the expressions of Kv3.4 mRNA and protein in rat PASMCs were significantly downregulated by 15-HETE (P<0.05). Based on all the information above, we conclude that Kv3.4 channel is involved in vasoconstriction induced by 15-HETE in rat PAs.
Animals
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Cells, Cultured
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Female
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Hydroxyeicosatetraenoic Acids
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pharmacology
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Hypertension, Pulmonary
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physiopathology
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Hypoxia
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physiopathology
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Male
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Muscle, Smooth, Vascular
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cytology
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pathology
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Pulmonary Artery
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cytology
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physiopathology
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RNA, Messenger
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genetics
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metabolism
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Rats
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Rats, Wistar
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Shaw Potassium Channels
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antagonists & inhibitors
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genetics
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metabolism
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Vasoconstriction
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drug effects
9.Effects of stable isotope labeled internal standard on determination of ivabradine and N-demethylivabradine in human plasma.
Dong-qin LIU ; Jing-hua YU ; Yi-fan ZHANG ; Da-fang ZHONG ; Ling HE ; Xiao-yan CHEN
Acta Pharmaceutica Sinica 2015;50(3):348-354
This study aims to develop a liquid chromatography with tandem mass spectrometry (LC-MS/MS) method for the simultaneous determination of ivabradine and N-demethylivabradine in human plasma, and investigate effects of stable isotope labeled (SIL) internal standard (IS) on ivabradine. The analytes and IS were extracted from plasma by protein precipitation with acetonitrile, and chromatographied on a Capcell PAK C18 (100 mm x 4.6 mm, 5 μm) column using a mobile phase of methanol and 5 mmol x L(-1) ammonium acetate. Multiple reaction monitoring with electrospray ionization (ESI) was used in the positive mode for mass spectrometric detection. The effect of ivabradine isotope peak [M+H+3] + on IS and the effect of SIL IS purity on ivabradine were evaluated. An appropriate concentration of SIL IS was chosen to permit method selectivity and linearity of the assay over the required range. The standard curves were demonstrated to be linear in the range of 0.100 to 60.0 ng x mL(-1) for ivabradine, and 0.050 0 to 20.0 ng x mL(-1) for N-demethylivabradine. The intra and inter day precision and accuracy were within the acceptable limits for all concentrations. Besides, the interaction between IS and ivabradine did not impact the determination of analytes. This method was successfully applied to a pharmacokinetic study of hydrogen sulfate ivabradine sustained release tablets on Chinese healthy volunteers.
Benzazepines
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blood
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Chromatography, High Pressure Liquid
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Delayed-Action Preparations
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Humans
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Isotope Labeling
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standards
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Reference Standards
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Reproducibility of Results
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Sensitivity and Specificity
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Spectrometry, Mass, Electrospray Ionization
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Tablets
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Tandem Mass Spectrometry
10.Therapeutic effects of FuZhiSan on Alzheimer's disease rat model:evaluation with PET imaging
Zhe, GUO ; Jin-ming, ZHANG ; Shu-lin, YAO ; Hui-ru, FENG ; Xu-ling, LI ; Da-yi, YIN ; Jia-he, TIAN
Chinese Journal of Nuclear Medicine 2010;30(4):246-250
Objective To assess the feasibility of using PET molecular imaging to evaluate the therapeutic effects of traditional Chinese medicine FuZhiSan (FZS) on the model of aging Alzheimer's disease (AD) rats. Methods Twenty aged AD rats (Sparague-Dawley rats,male) were randomly divided into FZS treated group (n = 10) and control group (n = 10). Another 10 healthy adult rats were as blank controls. Morris water maze record system was used for cognitive function assessment. Before and after FZS treatment 18 F-fluorodeoxyglucose (FDG) and 11 C-2- [4'-(methylamino) phenyl] benzothiazol-6-ol ( PIB )PET imaging was undertaken. After post-treatment imaging procedures the brain tissues of all animals were taken for histochemical study,such as staining with HE,congo red,amyloid β (Aβ) immunofluorescence,5-bromo-2-deoxyuridine (BrdU) immunofluorescence and NeuN immunofluorescence. Paired t-test was performed with SPSS 13.0 software for the data analysis. Results The cognitive dysfunction of aging AD rats was improved after FZS treatment. The escape latency in FZS treated group was significantly shorter than that of control group ((32.5 ±10.8) s vs (102.6±8.8) s,t =15.7987,P=0. 0001). Diffuse neuronal loss and Aβ deposition were detected in the hippocampus and cortex in the aged AD rats. The imaging data showed that brain glucose metabolism was amended in FZS treated group while the abatement of amyloid deposition was not significant. Immunofluorescence results indicated that the neuronal proliferation was more remarkable in FZS treated group. Conclusions It may be feasible to use PET imaging as a method to evaluate the therapeutic effect in AD rats. FZS may ameliorate memory dysfunction of aged AD rats. Its mechanism may be partly contributed to the enhancement of the neuronal proliferation and survival.