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Blocking Effect of Salvianolic Acid A on calcium channels in Isolated Rat Ventricular Myocytes

Chinese Journal of Integrative Medicine 2012年第5期18卷 366-370页

作者：王宝刘建勋孟红旭林成仁 The laboratory Research Center Xiyuan HospitalChina Academy of Chinese Medical Sciences

Objective： To study the effect of salvianolic acid A （SAA） on l-type calcium current （I-Cal） in isolated ventdcular myocytes of Sprague-Dawley rats. Methods： SPA powder was dissolved in normal Tyrode＇s solution to reach the concentrations of 1, 10, 100, and 1000 μmol/l. The traditional whole-cell patch-clamp recording technique was employed to evaluate the effects of SAA on I-Cal in single ventricular myocytes which were prepared by langendorff perfusion apparatus from Sprague-Dawley rats. Results： SPA （1, 10, 100, and 1000 μmol/l） inhibited I-Cal peak value by 16.23%± 1.3% （n=6, P〈0.05）, 22.9% ± 3.6% （n=6, P〈0.05）, 53.4% ± 3.0% （n=8, P〈0.01）, and 62.26% ± 2.9% （n=8, P〈0.01）, respectively. SAA reversibly inhibited I-Cal in a dose-dependent manner and with a half-blocking concentration （IC50） of 38.3 μmol/l. SAA at 100 μmol/l elevated the I-V curve obviously, and shifted the half-active voltage （V0.5） from （-15.78± 0.86） mV to （-11.24 ± 0.77） mV （n=6, P〈0.05） and the slope （K） from 5.33 ±0.74 to 4.35±0.74 （n=6, P〉0.05）. However, it did not alter the shapes of I-V curve, steady-state inactivation curve, or recovery from inactivation curve. Conclusions： SAA inhibited I-Cal in a dose-dependent manner. It shifted the steady-state activation curve to a more positive voltage, which indicated that the drug affected the activated state of calcium channels, and suggested that the Ca2. antagonistic effect of SPA be beneficial in the treatment of myocardial ischemia reperfusion injury.

关键词： salvianolic acid A ventricuiar myocytes patch-clamp technique l-type calcium channels

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Effect of resveratrol on l-type calcium current in rat ventricular myocytes

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Acta Pharmacologica Sinica 2006年第2期27卷 179-183页

作者： li-ping ZHANG Jing-xiang YIN Zheng lIU Yi ZHANG Qing-shan WANG~1 Juan ZHAO Department of Physiology,Institute of Basic Medicine,Hebei Medical University.Shijiazhuang 050017.China Department of Physiology Institute of Basic Medicine Hebei Medical University Shijiazhuang China

Aim:To study the effect of resveratrol on l-type calcium current(ICa-l)in isolatedrat ventricular myocytes and the mechanisms underlying these ***:ICa-lwas examined in isolated single rat ventricular myocytes by using the wholecell patch-clamp recording ***:Resveratrol(10-40 μmol/l)re-duced the peak amplitude of ICa-land shifted the current-voltage(I-V)curveupwards in a concentration-dependent ***(10,20,40 μmol/l)decreased the peak amplitude of ICa-lfrom -14.2±1.5 pA/pF to -10.5±1.5 pA/pF(Pl/l)shifted the steady-state activation curve of ICa-lto theright and changed the half-activation potential(V0.5from -19.4+0.4 mV to-15.4±1.9 mV(Pl at a concentration of 40 μmol/l did notaffect the steady-state inactivation curve of ICa-l,but did markedly shift the time-dependent recovery curve of ICa-lto the right,and slow down the recovery ofICa-lfrom *** orthovanadate(Na3VO4;1 mmol/l),a potentinhibitor of tyrosine phosphatase,significantly inhibited the effects of resveratrol(Plusion:Resveratrol inhibited ICa-lmainly by inhibiting the activa-tion of l-type calcium channels and slowing down the recovery of l-type cal-cium channels from *** inhibitory effect of resveratrol was medi-ated by the inhibition of protein tyrosine kinase in rat ventricular myocytes.

关键词： resveratrol patch-clamp techniques myocardium l-type calcium channels

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Comparative identification of Ca～(2+) channel expression in INS-1 and rat pancreatic β cells

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World Journal of Gastroenterology 2009年第24期15卷 3046-3050页

作者： Fei li Zong-Ming Zhang Department of General Surgery Digestive Medical Center The First Affiliated Hospital Medical School Tsinghua University Beijing 100016 China

AIM： To identify and compare the profile of Ca^2＋ channel subunit expression in INS-1 and rat pancreatic β cells.METHODS： The rat insulin-secreting INS-1 cell line was cultured in RPMI-1640 with Wistar rats employed as islet donors. Ca^2＋ channel subunit expression in INS-1 and isolated rat β cells were examined by reverse transcription polymerase chain reaction （RT-PCR）. Absolute real-time quantitative PCR was performed in a Bio-Rad iQ5 Gradient Real Time PCR system and the data analyzed using an iQ5 system to identify the expression level of the Ca^2＋ channel subunits. RESUlTS： In INS-1 cells, the l-type Ca^2＋ channel 1C subunit had the highest expression level and the TPRM2 subunit had the second highest expression. In rat β cells, the TPRC4β subunit expression was dominant and the expression of the l-type lC subunit exceeded the 1D subunit expression about two-fold. This result agreed with other studies, confirming the important role of the l-type lC subunit in insulinsecreting cells, and suggested that non-voltage-operated Ca^2＋ channels may have an important role in biphasic insulin secretion. CONClUSION： Twelve major Ca^2＋ channel subunit types were identified in INS-1 and rat β cells and significant differences were observed in the expression of certain subunits between these cells.

关键词： l-type calcium channels Expression profile Insulin-secreting cells Rats pancreatic β cell Reverse transcription-polymerase chain reaction

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Effects of AMP579 and adenosine on l-type Ca2+current in isolated rat ventricular myocytes

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Acta Pharmacologica Sinica 2005年第5期26卷 559-562页

作者： Xiong WANG~1,Bo-wei WU,Dong-mei WUDepartment of Physiology Shanxi Medical University,Taiyuan 030001,China Department of Physiology Shanxi Medical University Taiyuan China

Aim:To compare the effects of AMP579 and adenosine on l-type Ca2+current(ICa-l)in rat ventricular myocytes and explore the mechanism by which AMP579acts on ***:ICalwas recorded by patch-clamp technique in ***:Adenosine(10nmol/l to 50μmol/l)showed no effect onbasal ICal,but it inhibited the ICalinduced by isoproterenol 10 nmol/l in a concen-tration-dependent manner with the IC50of 13.06 μmol/*** to adenosine,AMP579 also showed an inhibitory effect on the ICalinduced by ***579 and adenosine(both in 10 μmol/l)suppressed isoproterenol-inducedICalby 11.1% and 5.2%, *** addition,AMP579 had a direct inhibitoryeffect on basal ICalin a concentration-dependent manner with IC50(1.17 μmol/l).PD 116948(30μmol/l),an adenosine A1receptor blocker,showed no action on theinhibitory effect of AMP579 on basal ***,GF109203X(0.4μmol/l),aspecial protein kinase C(PKC)blocker,could abolish the inhibitory effect of AMP579on basal I*** the inhibitory effect of AMP579 on basal ICalwas inducedthrough activating PKC,but not linked to adenosine ***:AMP579 shows a stronger inhibitory effect than adenosine on the ICalinducedby ***579 also has a strong inhibitory effect on basal ICalin ratventricular *** of PKC is involved in the inhibitory effect ofAMP579 on basal ICalat downstream-mechanism.

关键词： AMP579 adenosine heart ventricles cardiac myocytes l-type calcium channels patch-clamp techniques

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TGF-β1 regulates P-JNK and l-type calcium Channel Cav1.2 in cortical neurons

TGF-β1 regulates P-JNK and L-Type Calcium Channel Cav1.2 in...

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第十二届全国免疫学学术大会

作者： liu Zhenning sheng Jiangtao Peng Guoyi Yang Jinhua Chen Weiqiang li Kangsheng Department of Microbiology and Immunology Key Immunopathology Laboratory of Guangdong ProvinceShantou University Medical College Department of Neurosurgery First Affiliated HospitalShantou University Medical College

Background:Previous studies showed that central nervous system(CNS) diseases could cause a series of neuronal lesions,which may be improved by the anti-apoptotic neuroprotection of transforming growth factor-beta1(TGF-β1).In neurons,l-type Ca channels(lTCC) are mainly composed of Cavl.2 ***:Given that TGF-βi is implicated in numerous CNS diseases,we examined the neuroprotective effects of TGF-β1 on the Cav1.2 channel in the ***:To simulate acute mechanical traumatic brain injury(TBI),we used a needle to create parallel scratches across culture plates for 9 ***4-AM loaded,dual wavelength of 488 nm/530 nm in laser scanning confocal microscopy was used to determine intracellular calcium concentrations([Ca]).Results:We found that MAPK inhibitors impede the TGF-β1-induced cell viability and TGF-β1 recovered the traumainduced cell viability in ***.2 was significantly decreased in the TGF-β1-treated(10 ng/ml) *** this TGF-β1 concentration,Cavl.2 was significantly down-regulated in a time-dependent manner after 12 ***,TGF-β1 partially recovered the protein level of Cavl.2 reduced by ***-β1 significantly inhibits the fluorescence intensity of[Ca] increased by KC1 and delays the time of peak[Ca].The observed effects of TGF-β1 on Cav1.2 were regulated by MAPK *** observed effects of TGF-β1 on P-JNK also were impeded by pre-incubation with lTCC inhibitor(10 μM) nimodipine in trauma injured ***:Altogether,TGF-β1 regulates lTCCs in a mechanism dependent on the MEK,JNK1/2 and p38 MAPK signal pathways in cortical neurons,and we suggest this mechanism involved in cell viability.

关键词： Cortical neurons Jun NH2-terminal kinase 1/2 l-type calcium channels Transforming growth factor-b

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