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Effects of sodium β-aescin on expression of adhesion molecules and migration of neutrophils after middle cerebral artery occlusion in rats

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Acta Pharmacologica Sinica 2004年第7期25卷 23-29页

作者： Xia-min HU, Yan ZHANG2.Fan-dian ZENG3. Instituteof Clinical pharmacology 2.epartment of Anatomy, TongjiMedical College of {3. {2.of Science and Technology, wuhan 43.03., china Institute of Clinical pharmacology Department of Anatomy Tongji Medical College of Huazhong University of Science and Technology

AIM: To investigate the effects of sodium β-aescin on neutrophil migration and expression of adhesion molecules (ICAM-1 and E-selectin) after middle cerebral artery occlusion (MCAO) in rats. METHODS: Rats were pretreated with sodium β-aescin for 7 d and then subjected to cerebral ischemia/reperfusion (I/R) injury induced by an MCAO. After a 2.h ischemia and a 2.-h reperfusion, the infarct volume and neurological deficit were determined by the method of TTC staining and the Longa’s score. The effect of sodium β-aescin on the migration of neutro- phils was evaluated by measuring the activity of myeloperoxidase (MPO) enzyme. The expressions of adhesion molecules were determined by immunohistochemistry and Western blot. RESULTS: Sodium β-aescin significantly reduced the cerebral infarct volume and ameliorated the neurological deficit (P<0.05 or P<0.01). The MPO activity and the expressions of ICAM-1 and E-selectin in the vehicle-treated rats were increased significantly (P<0.01) after cerebral I/R. After treatment with sodium β-aescin, the enzymatic activity of MPO and the expressions of these adhesion molecules were significantly reduced compared with the vehicle-treated group (P<0.05 or P<0.01). CONCLUSION: Sodium β-aescin can attenuate brain injury, down-regulate the protein expressions of ICAM-1 and E-selectin, and reduce the migration of neutrophils after cerebral I/R.

关键词： brain ischemia aescin neutrophils peroxides intercellular adhesion molecule-1 E-selectin

Minocycline inhibits 5-lipoxygenase activation and brain inflammation after focal cerebral ischemia in rats

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Acta Pharmacologica Sinica 2007年第6期28卷 763-772页

作者： Li-sheng CHU~(2.3.,San-hua FANG~2.Yu ZHOU~2.Guo-liang YU~2.Meng-ling WANG~2.Wei-ping ZHANG~2.Er-qing WEI~(2.4)~2.Department of pharmacology,School of Medicine,Zhejiang university,Hangzhou 3.0058,{2. ~3.Department of Physiology ZhejiangChinese Medical university Hangzhou 3.0053.{2. Department of pharmacology School of Medicine Zhejiang University Hangzhou 310058 China

Aim:To determine whether the anti-inflammatory effect of minocycline on postis-chemic brain injury is mediated by the inhibition of 5-lipoxygenase （5-LOX）expression and enzymatic activation in ***:Focal cerebral ischemiawas induced for 3. min with middle cerebral artery occlusion,followed *** ischemic injuries,endogenous IgG exudation,the accumulationof neutrophils and macrophage/microglia,and 5-LOX mRNA expression weredetermined 72.h after reperfusion.5-LOX metabolites （leukotriene B4and cysteinylleukotrienes） were measured 3.h after ***:Minocycline （2..5and 45 mg/kg,ip,for 3.d） attenuated ischemic injuries,IgG exudation,and theaccumulation of neutrophils and macrophage/microglia 72.h after *** inhibited 5-LOX expression 72.h after reperfusion and the production ofleukotrienes 3.h after reperfusion,Conclusion:Minocycline inhibited postis-chemic brain inflammation,which might be partly mediated by the inhibition of5-LOX expression and enzymatic activation.

关键词： minocycline 5-1ipoxygenase leukotriene focal cerebral ischemia inflammation

Rapid quantification of the metabolite of valacyclovir hydrochloride in human plasma by liquid chromatography-tandem mass spectrometry

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Journal of Pharmaceutical Analysis 2010年第2期22卷 83-90页

作者： Yuan Tian1,2.Hui Lin1,2.Xue-Yu Zhang1,2.Zun-Jian Zhang1,2.Guo-Guang Mao3.. Key Laboratory of Drug Quality Control and Pharmacovigilance (china Pharmaceutical university),Ministry of Education,Nanjing 2.0009 2. Center for Instrumental Analysis,china Pharmaceutical university,Nanjing 2.0009 3. Department of Clinical pharmacology,Wannan Medical College,Wuhu 2.1000,china. Key Laboratory of Drug Quality Control and Pharmacovigilance (china Pharmaceutical university) Ministry of Education Nanjing 210009 China Center for Instrumental Analysis China Pharmaceutical University Nanjing 210009 China Department of Clinical pharmacology Wannan Medical College Wuhu 241000 China

Objective To establish a rapid,sensitive and selective liquid chromatography-tandem mass spectrometry (LC-MS/MS) method for the determination of acyclovir (the metabolite of valacyclovir hydrochloride) in human plasma. Methods After addition of ganciclovir as internal standard (IS),plasma samples were prepared by one-step protein precipitation using acetonitrile as precipitant,followed by an isocratic elution with 0.1% formic acid solution-methanol (95∶5,v/v) on an Agilent ZORBAX SB-C18 (150mm×2.1mm i.d.,3.5μm) column. Detection was performed on a triple-quadrupole mass spectrometer utilizing electrospray ionization (ESI) interface operating in positive ion and selected reaction monitoring (SRM) mode with the precursor to product ion transitions m/z 2.6.2.152.1 for acyclovir and m/z 2.6.2.152.1 for the IS. Results The analytical results demonstrated a good linearity over the ranges from 0.005 to 4μg/mL (r=0.9999) for valacyclovir hydrochloride. The relative standard deviations (RSD) of intra-batch and inter-batch were less than 4.06% and 9.2.%,respectively. The limit of detection and lower limit of quantification in human plasma were 2.g/mL and 5ng/mL,respectively. Conclusion The method was simple,sensitive,accurate and reproducible and has been successfully applied to a bioequivalence study of valacyclovir hydrochloride capsules in Chinese healthy male volunteers.

关键词： valacyclovir hydrochloride acyclovir liquid chromatography-tandem mass spectrometry method validation human plasma

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Circadian expression of clock genes and angiotensin Ⅱ type 1 receptors in suprachiasmatic nuclei of sinoaortic-denervated rats

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Acta Pharmacologica Sinica 2007年第4期28卷 484-492页

作者： Hui LI~2.Nmg-ling SUN~(2.4),Jin WANG~3.Ai-jun LIU~3.Ding-feng SU~(3.4)~2.epartment of Cardiology,Peking {3. People's Hospital,Beijing 100044,china ~3.epartment of {2.,Second MilitaryMedical {3.,Shanghai 2.043.,china Department of Cardiology Peking University People's Hospital Beijing China Department of pharmacology Second Military Medical University Shanghai China

Aim:To investigate whether the circadian expression of central clock genes andangiotensin Ⅱ type 1 (AT1) receptors was altered in sinoaortic-denervated (SAD)***:Male Sprague-Dawley rats underwent sinoaortic denervation or asham operation at the age of 12.*** weeks after the operation,bloodpressure and heart period were measured in the conscious state in a group ofsham-operated (n=10) and SAD rats (n=9).Rest SAD and sham-operated ratswere divided into 6 groups (n=6 in each group).The suprachiasmatic nuclei (SCN)tissues were taken every 4 h throughout the day from each group for the determi-nation of the mRNA expression of clock genes (Per2.and Bmal1) and the AT1receptor by RT-PCR;the protein expression of Per2.and Bmal1 was determined byWestern ***:Blood pressure levels in the SAD rats were similar tothose of the sham-operated ***,blood pressure variabilities signifi-cantly increased in the SAD rats compared with the sham-operated *** variation of clock genes in the SCN of the sham-operated rats was char-acterized by a marked increase in the mRNA and protein expression during ***2.and Bmal1 mRNA levels were significantly lower in the SAD rats,especially during dark *** blot analysis confirmed an attenuation ofthe circadian rhythm of the 2.clock proteins in the SCN of the SAD ***1receptor mRNA expressions in the SCN were abnormally upregulated in the lightphase,changed to a 12.h cycle in the SAD ***:The circadian varia-tion of the 2.central clock genes was attenuated in the SAD *** baroreflexdysfunction also induced a disturbance in the expression of AT1 receptors in theSCN.

关键词： circadian rhythm arterial baroreflex clock genes sinoaortic denervation reninangiotensin system

Inhibition of β-lactamase-mediated oxacillin resistance in Staphylococcus aureus by a deoxyribozyme

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Acta Pharmacologica Sinica 2007年第11期28卷 1775-1782页

作者： Zheng HOU~2.Jing-ru MENG~2.Jin-rong ZHAO~3.Ben-quan HU~2.Jie LIU~2.Xiao-jun YAN~3.Min JIA~2.Xiao-xing LUO~(2.4)~2.Department of pharmacology and ~3.Institute of Genetic Diagnosis,The Fourth Military Medical university,Xi’an 71003.0}{2. Department of pharmacology The Fourth Military Medical University Xi'an 710032 China Institute of Genetic Diagnosis The Fourth Military Medical University Xi'an 710032 China

Aim:To investigate the oxacillin susceptibility restoration of methicillin-resistantStaphylococcus aureus (MRSA) by targeting the signaling pathway of blaR1-blaZ with a ***:ADNAzyme (named PS-DRz602. targeting blaR1mRNA was designed and *** DRz602.was introduced into a MRSAstrain WHO-2.the colony-forming units of WHO-2.on the Mueller-Hinton agarcontaining 6 mg/L oxacillin and the minimum inhibitory concentrations of oxacillinwere *** inhibitory effects of DRz602.on the expressions of antibi-otic-resistant gene blaR1 and its downstream gene blaZ were detected by realtime ***:PS-DRz602.significantly decreased the transcription ofblaR1 mRNA and led to the significant reduction of blaZ in a concentration-dependent ***,the resistance of S aureus WHO-2.to the β-lactam antibiotic oxacillin was significantly ***:Our resultsindicated that blocking the blaR1-blaZ signaling pathway via DNAzyme mightprovide a viable strategy for inhibiting the resistance of MRSA to β-lactam antibi-otics and that BlaR1 might be a potential target for pharmacological agents com-bating MRSA.

关键词： methicillin-resistant Staphylococcus aureus BlaR1 phosphorothioate deoxyribozyme antibiotic resistance

Propranolol and verapamil inhibit mRNA expression of RyR2.and SERCA in L-thyroxin-induced rat ventricular hypertrophy

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Acta Pharmacologica Sinica 2004年第03期25卷 93-97页

作者： rat ventricular hypertrophy1 Xiao-dong WU2.De-zai DAI2.Qiu-pin ZHANG3.Feng GAO3.Research Division of pharmacology,{3. Pharmaceutical {2. 3.aboratorial Center ofMedicine,Southeast {2.,Nanjing 2.0009,{3. Research Division of pharmacology China Pharmaceutical University Laboratorial Center of Medicine Southeast University

AIM: To study the alteration in the mRNA level of cardiac ryanodine receptor 2.(RyR2. and sarco-endoplasmic reticulum Ca2.-ATPase (SERCA) in L-thyroxin-induced hypertrophy. METHODS: L-thyroxin (500 g/kg) daily was injected for 10 d. RT-PCR was used to determine mRNA expression. RESULTS: An increase in the relative amount of RyR2.(111 %) and SERCA mRNA (65 %) expression was observed in the hypertrophied rats (RyR2. 77±11; SERCA: 87±10, n=9) compared with the normal rats (RyR2. 3.±10; SERCA: 53.10, n=9). Propranolol was effective to inhibit the increase in RyR2.(51±7) and SERCA (63.13. mRNA expression in hypertrophied rats, respectively. Verapamil also reduced RyR2.(62.5) and SERCA (75±8) mRNA expression. CONCLUSION: Both RyR2.and SERCA mRNA level in L-thyroxin-induced cardiac hypertrophy was over-expressed and propranolol or verapamil inhibited the alteration.

关键词： heart hypertrophy arrhythmia ryanodine receptors Ca2.-transporting ATPase L-thyroxin propranolol verapamil reversed transcriptase polymerase chain reaction

Inhibitory effect of leflunomide on hepatic fibrosis induced by CCl4 in rats

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Acta Pharmacologica Sinica 2004年第7期25卷 69-74页

作者： Hong-wei YAO2.3. Jun LI3.4, Ji-qiang CHEN2. Shu-yun XU3. 2.Zhejiang Respiratory Drugs Research Laboratory of State Drugs Administration of{3., School of Medicine, Zhejiang university, Hangzhou 3.003. 3.nstitute of Clinical {2., Anhui Medical university, Hefei 2.003.0}{3. Zhejiang Respiratory Drugs Research Laboratory of State Drugs Administration of china School of Medicine Zhejiang University Hangzhou 310031 Chin

AIM: To study the effect of leflunomide on CCl4-induced hepatic fibrosis in rats. METHODS: Hepatic fibrosis was induced by subcutaneous injection with 50 % CCl4 in Sprague-Dawley rats. The amount of CCl4 administered was 1 mg/kg. The alanine aminotransferase (ALT), aspartate aminotransferase (AST), nitric oxide (NO) levels in plasma and hydroxyproline (Hyp) contents in liver tissue were assayed by spectrophotometry. The hyaluronic acid (HA) and procollagen III (PC III) were assessed by radioimmunoassay. The transforming growth factor-β1 (TGF-β1) in serum was determined by ELISA. The nuclear factor-kappa B (NF-κB) in liver tissue was examined by immunohistochemistry. Liver samples collected after 12.weeks of CCl4 treatment were stained with hematoxy- lin and eosin. RESULTS: Leflunomide (1, 3. and 9 mg/kg) significantly decreased indices of liver and spleen, the serum transaminase (AST, ALT) activities, HA and PC III levels, and Hyp contents in liver tissue in rats of hepatic fibrosis. Histopathological examination showed leflunomide had inhibitory effect on fibrogenesis and formation of pseudolobulus. Furthermore, leflunomide significantly inhibited NF-κB expression in liver tissue, and reduced elevated serum TGF-β1 and NO levels in rats of hepatic fibrosis. CONCLUSION: Leflunomide showed inhibitory action on hepatic fibrosis induced by CCl4 in rats.

关键词： leflunomide hepatic fibrosis nuclear factor-kappa B transforming growth factor-beta nitric oxide

Dispersion of ventricular mRNA of RyR2.and SERCA2.associated with arrhythmogenesis in rats

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Acta Pharmacologica Sinica 2004年第6期25卷 36-41页

作者： Hong-lan WANG, De-zai DAI2. Feng GAO3. Yan-pin ZHANG3. Feng LU Research Division of pharmacology, china Pharmaceutical {2., Nanjing 2.0009, china 3.aboratorial Center ofMedicine, Southeast {2., Nanjing 2.0009, china Research Division of pharmacology China Pharmaceutical University Nanjing 210009 China

AIM: To investigate the effect of CPU86017 on the changes of mRNA abundance of different calcium handling system in infarcted heart. METHODS: Rats were subjected to left coronary ligation to induce myocardial infarc- tion (MI). The treatment with either propranolol (Pro) 5 mg/kg ip or CPU86017 1, 2. and 4 mg/kg ip was initiated on the next day of operation and continued for 2. d. Medication with isoproterenol (Isop) 3.mg/kg sc started on the d 17-2.. Ventricular mRNA abundance of ryanodine receptor 2.(RyR2., sarcoplasmic reticulum Ca2.-ATPase (SERCA2., L-type Ca2. channel, and Na+/Ca2.exchanger (NCX1) were measured. RESULTS: Arrhythmic scores (AS) in the Isop group was raised up to 5.2.±1.75 (P2.2.±2.04 and sham group 1.50±1.73. The AS was depressed by Pro (1.63.1.53. P2.and 4 mg/kg (3.00±1.2., and 1.70±1.85, P2. and SERCA2.was associated with an increase in AS in Isop group, and it was much depressed in the left than the right ventricle. The dispersion and depression of mRNA were restored significantly by Pro and CPU86017, asso- ciated with suppression on AS. In Isop group, the mRNA abundance of L-type Ca2. channel was not changed; and a moderate increase in the mRNA of NCX1 was seen, the changes were regressed by Pro and CPU86017. CONCLUSION: Isop-induced arrhythmogenesis in MI heart was correlated mainly with a dispersion of depressed mRNA abundance in ventricle likely due to the consequence of PKA over-phosphorylation. A suppression of arrhythmia by Pro and CPU86017 resulted from a regression of the dispersion and depression of RyR2.and SERCA2.

关键词： myocardial infarction isoproterenol arrhythmia calcium 4-chlorobenzyltetrahydroberberine

Alterations of placental cytochrome P450 1A1 and P-glycoprotein in tobacco-induced intrauterine growth retardation in rats

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Acta Pharmacologica Sinica 2005年第11期26卷 1387-1394页

作者： You-e YAN~2. Hui WANG~(2.3., Ymg-hong FENG~2.~2.Department of pharmacology, Medical College of wuhan {2., wuhan 43.071, china Departmentofpharmacology MedicalCollegeofWuhanUniversityWuhan430071China

Aim: To investigate the alterations of placental P-glycoprotein (P-gp) and cytochrome P450 1 Al (CYPlAl) at different gestational days (GD), and to explore the possible significance of placental P-gp and CYPlAl in tobacco smoke-induced intrauterine growth retardation (IUGR) in rats. Methods: An IUGR model was produced by passive tobacco smoking from GD 7 to parturition (GD 2.) and predicted using fetal development parameters. Placental structure and function were monitored by observing pathological alteration and antioxidative function, including the content of malondialdehyde and the activities of superoxide dismutase and catalase (CAT). The expressions of CYPlAl and P-gp (mdr la and mdr lb) were detected using a reverse transcription polymerase chain reaction and immunohistochemistry. Results: Placental pathological changes occurred and the malondialdehyde content increased, whereas the activities of superoxide dismutase and CAT lowered, when compared to their controls. In the rat placenta of the tobacco group, the level of CYPlAl mRNA increased significantly; the level of mdr1a mRNA increased significantly at GD 2. but not at GD 14, whereas the level of mdr1b mRNA in different term remained stable; the expression of P-gp increased significantly only in full-term placenta. Conclusion: The expression of placental CYPlAl and P-gp increased in tobacco-induced IUGR. Overexpression of placental CYPlAl can attribute to the metabolism of tobacco and the generation of reactive metabolites, which can trigger IUGR. As a compulsory mechanism, upregulation of P-gp might decrease tobacco exposure to a developing fetus with IUGR

关键词： intrauterine growth retardation tobacco placenta cytochrome P450 1A1 P-glycoprotein rats