咪达唑仑对hERG钾通道的作用

目的:观察咪达唑仑对人胚肾上皮细胞(HEK-293)中异源表达的人类相关基因(h ERG)钾电流作用及其机制。方法:利用全细胞膜片钳技术,观察咪达唑仑对h ERG钾通道的抑制作用,分析其对通道激活、失活动力学过程的影响以及咪达唑仑对Y652A和F656C突变型h ERG钾通道的作用。结果:咪达唑仑浓度依赖性地抑制h ERG钾电流,其IC50值为(1.31±0.32)μmol/L。1.0μmol/L的咪达唑仑加药前后半数激活电压V1/2由(2.32±0.38)m V变为(-1.96±0.83)m V;加药前后半数失活电压V1/2由(-49.25±0.69)m V变为(-57.53±0.53)m V(P0.05),失活曲线左移;与野生型(WT)比较,Y652A和F656C突变型可显著减弱咪达唑仑对h ERG通道的阻断作用。结论:咪达唑仑能阻断h ERG钾通道,失活速度加快,Y652和F656可能是咪达唑仑与h ERG钾通道结合的关键位点。     

钩藤碱对human ether-a-go-go相关基因通道的抑制作用

将human ether-a-go-go相关基因(HERG)cRNA注射到非洲爪蟾卵母细胞,采用双电极电压钳技术,观察钩藤碱对表达电流的影响.结果显示(1)钩藤碱抑制HERG通道的表达是浓度依赖性的,IC50为(773.4±42.5)μmol/L.(2)钩藤碱抑制HERG通道的表达是电压依赖性的,最大抑制率在-20 mV,为15%.上述结果提示,钩藤碱抑制HERG编码的钾通道,导致心室复极时间延长,揭示了与钩藤碱相关的心肌钾通道的分子生物学基础.     

他莫昔芬对SHG-44人胶质瘤细胞钠通道的抑制作用

目的：研究他莫昔芬对SHG-44胶质瘤细胞钠通道电流的作用。方法：采用全细胞膜片钳方法记录SHG-44细胞的钠通道电流,并观察施用不同浓度的他莫昔芬后电流的变化。结果：该钠通道电流特性为内向电流、快速激活失活,他莫昔芬能够明显阻断该电流,该阻断具有剂量依赖性及电压依赖性。在0mV时,8μmol／L他莫昔芬对钾电流抑制率为69％。半数抑制浓度（IC50）为5．54μmol／L。结论：他莫昔芬可明显阻断SHG-44胶质瘤细胞上的钠通道,这可能是他莫昔芬抑制胶质瘤细胞增殖的机制之一。     

嗜热酯酶APE1547催化活性的定向进化研究

对来源于嗜热古菌Aeropyrum pernix的酯酶(APE1547)催化活性进行定向进化研究。利用APE1547特殊的稳定性,建立了准确的高通量高温酯酶筛选方法。对第一代随机突变库筛选获得了催化活性较野生型提高1.5倍的突变体M010,序列分析表明其氨基酸突变为R526S。从第二代突变库中筛选出的总活力提高5.8倍突变体M020,突变位点为R526S/E88G/A200T/I519L,其比活力与M010一致,但表达量比野生型提高约4倍。对M020酶学性质表征发现,其最适pH为8.5,比野生型向碱性偏移0.5;活性中心残基酸性基团的解离常数(pK1)由野生型的7.0提高至7.5。晶体结构分析表明,突变位点R526距离活性中心较近,将其突变为Ser降低了活性中心的极性,抑制了催化残基His的解离,使酸性基团的解离常数升高。     

多不饱和脂肪酸对成年雪貂心肌钾通道的作用

本研究是在成年雪貂的心肌上研究多不饱和脂肪酸（PUFA）对电压门控钾通道的效应。我们观察到,n-3 PUFA能抑制短时性外向钾电流（Ito)和延迟整流钾电流（IK),而对内向整流钾电流（IK1）则没有明显影响。二十二碳六烯酸（DHA）对Ito和Ik能产生浓度依赖性的抑制作用,其IC50分别为7.5和20μmol/L,但不影响IK1。二十碳五烯酸（EPA）对这三种钾通道的作用与DHA相似。花生四烯酸（5或10μmol/L)先引起IK的抑制,然后引起IK,AA的激活;用环氧合酶抑制剂消炎痛可以阻断花生四烯酸激活IK,AA的作用。不具有抗心律失常作用的单不饱和脂肪酸和饱和脂肪酸都不明显影响这些钾通道的活性。上述实验结果证明,n-3 PUFA能抑制心肌细胞的Ito和IK,但和我们以前报道的PUFA对心肌钠电流和钙电流的作用相比,其对Ito和IK抑制作用的效能较低。n-3 PUFA的抗心律失常效应可能与它们抑制心肌钠、钙、钾通道的作用有关。     

HERG钾通道在细胞容量调节中的作用

目的:研究HERG钾通道在细胞容量调节中的作用,并探讨其作用机制。方法:全部试验应用稳定转染HERG-HEK293细胞和HEK293细胞。应用全细胞膜片钳技术记录HERG钾电流。结果:1当HERG-HEK293细胞处于低渗状态,指令电压为0 mV时,Istep增加60%(n=12,P0.05);如指令电压为+30 mV,Itail增加72.1%(n=11,P0.01),增大的HERG电流可被特异性HERG钾电流阻断剂Cisapride(100nM)抑制,Istep被抑制97.2%(n=6,P0.01),Itail被抑制174.1%(n=6,P0.01)。2在低渗状态,指令电压为0 mV时,容量调节性氯通道阻断剂尼氟灭酸(NFA,10 nmol/L)使Istep抑制46.2%(n=12,P0.01),Itail抑制48.5%(n=11,P0.01);给以容量调节性氯通道阻断剂DIDS 100μmol/L时,Istep抑制45.9%(n=12,P0.01),Itail抑制51.1%(n=11,P0.01)。结论:HERG通道部分参与调节性细胞容量下降过程。其参与的细胞容量调节与容量调节性氯通道的激活相伴随。     

简单节杆菌3-甾酮-△1 -脱氢酶基因的克隆表达及定点突变研究

目的:将来源于简单节杆菌的3-甾酮-△~1-脱氢酶(3-ketosteroid-Delta(1)-dehydrogenase,KSDD)在大肠杆菌中进行表达,获得具有活性的脱氢酶;利用计算机预测KSDD的三级结构,并通过定点突变确定酶的关键位点以期优化脱氢酶的活性及性质。方法:克隆简单节杆菌编码KSDD的基因ksdd构建原核表达载体,以Escherichia coli BL21(DE3)为表达宿主构建重组菌并诱导表达,HPLC法检测重组酶催化4-AD脱氢的转化率;通过SWISS-MODEL同源建模分析KSDD结构,对预测的催化关键位点氨基酸残基进行定点突变并研究突变后重组酶的活性变化。结果:成功构建了表达脱氢酶KSDD的重组菌E.coli pET-22-ksdd,21℃下诱导表达后,重组酶对4-AD的转化率为27%;通过SWISS-MODEL同源建模预测出脱氢酶结构并对4个关键位点进行定点突变设计,获得突变子Y120R、Y320L、Y488F和G492Y。突变子Y120R和Y488F失活,证明其为酶的活性位点;突变子Y320L的转化率与野生型基本一致,但37℃反应条件下稳定性有所提高;突变子G492Y对4-AD的转化率是野生型的1.2倍,37℃条件下稳定性有所提高,是突变后氨基酸位点疏水性增加和周围静电作用改变所导致。结论:目前对简单节杆菌3-甾酮-△~1-脱氢酶结构分析及催化机理相关的研究较少,本研究验证了KSDD的活性位点,优化了酶的稳定性,为进一步对酶的性质进行定向改造打下了基础。     

阻断炎症部位5-HT2A受体对大鼠完全弗氏佐剂诱发炎性痛和脊髓背角神经肽Y表达的作用

炎症时组织释放5-羟色胺(5-hydroxytryptamine,5-HT),对炎性痛觉过敏的产生起重要作用。但炎症组织5-HT2A受体在其中的意义尚不明确。本文旨在研究外周组织5-HT2A受体在慢性炎性痛中的作用。大鼠后足底注射完全弗氏佐剂(complete Freund’s adjuvant,CFA),在局部炎性部位给予选择性5-HT2A受体阻断剂酮色林,用行为学检测后足底对热和机械刺激的撤足反射,用免疫组织化学方法检查脊髓背角和背根神经节(dorsal root ganglion,DRG)中神经肽Y(neuropeptide Y,NPY)表达变化。结果显示,炎症局部给予酮色林(20、40、80μg)能剂量依赖性地抑制CFA诱发的热痛觉过敏;每日给予80μg酮色林,在第三天即完全翻转CFA引起的热痛觉过敏,以及部分地减轻触觉超敏。CFA诱发脊髓背角I~II层NPY表达增加,炎症组织局部注射酮色林能抑制脊髓背角NPY的表达增加,但不改变DRG中NPY的表达。这些结果表明:外周炎症组织5-HT2A受体激活参与慢性痛觉过敏的发生;阻断炎症部位5-HT2A受体能缓解疼痛,矫正与病理疼痛密切关联的脊髓背角细胞异常改变。因此,外周5-HT2A受体,有望成为治疗慢性炎性痛觉敏感性增高、不产生中枢神经系统副作用的药物靶点。     

Myxococcus sp.V11海藻糖合酶TreS II分子改造 *

来源于黏细菌Myxococcus sp.V11的海藻糖合酶(trehalose synthase, EC 2.4.1.245)TreS II可通过转糖苷作用将麦芽糖转化成为海藻糖,在酶法生产海藻糖上显示出一定的应用潜力,但TreS II对热敏感,在60℃保温3h,酶活性丧失,限制了其应用范围.目的:拟探索TreS II影响热稳定性的氨基酸残基构成,通过对可能的氨基酸位点进行定点突变,以期获得耐热性的突变子,扩大TreS II应用范围.方法: 通过PCR介导的方法对TreS II可能影响到热稳定性的氨基酸Q3,A283,W374,R449和Y537进行定点突变,以野生型重组酶为对照,比较突变型与野生型的最适反应温度和最适反应pH,通过测定不同温度下保存不同时间后的残留酶活,检测突变子的耐热效果.结果: 研究表明突变子Q3D,A283R,W374D,R449Q和Y537H的比酶活与野生型无显著差异,且最适pH 和最适反应温度也未发生改变;A283R,Y537H在60℃条件下,3h后活性剩余68%;Q3D,W374D,R449Q在温度60℃时,3h后活性剩余35%.结论: TreS II分子结构中与金属离子结合的几个氨基酸残基的改变对蛋白质分子的耐热性具有显著影响.     

HERG通道在爪蟾卵母细胞的持续表达与电流变化

目的:探索一个HERG通道在爪蟾卵母细胞持续稳定表达HERG通道的方法,研究表达培养不同时期卵母细胞膜静息电位和通道电流特性的变化.方法:用含HERG片段的pSP64载体质粒体外转录制备的mRNA注射表达于卵母细胞,用双电极电压钳技术记录通道电流.结果:①本方法可成功在卵母细胞持续表达与HERG通道电生理特性一致的功能性通道,并可在10～15 d内稳定用于电生理记录.②在培养的第3、6和9 d细胞膜静息电位负值逐渐升高,在第12 d又开始降低.异源表达的HERG通道内向整流的最大电位在第3、6和9 d逐渐向正向转移,在第12 d又回复,激活曲线的半最大激活电压(V1/2)在培养的第3、6和9 d逐渐向负向转移,在第12 d又逐渐回复,其改变和膜静息电位的变化趋势一致.结论:本研究提供了一种HERG基因在爪蟾卵母细胞长期持续表达的方法,并为HERG通道的分子位点和药物作用研究在不同实验条件电生理实验数据的差异提供依据.     

High affinity HERG K(+) channel blockade by the antiarrhythmic agent dronedarone: resistance to mutations of the S6 residues Y652 and F656

Pharmacological inhibition of human-ether-a-go-go-related gene (HERG) K(+) channels by structurally and therapeutically diverse drugs is associated with the 'acquired' form of long QT syndrome and with potentially lethal cardiac arrhythmias. Two aromatic amino-acid residues (Y652 and F656) on the inner (S6) helices are considered to be key constituents of a high affinity drug binding site within the HERG channel pore cavity. Using wild-type (WT) and mutant HERG channels expressed in mammalian cell lines, we have investigated HERG channel current (I(HERG)) blockade at 37+/-1 degrees C by dronedarone (DRONED), a non-iodinated analogue of the Class III antiarrhythmic agent amiodarone (AMIOD). Under our conditions WT I(HERG) tails, measured at -40 mV following activating pulses to +30 mV, were blocked with IC(50) values of approximately 59 and 70 nM for DRONED and AMIOD, respectively. I(HERG) inhibition by DRONED was contingent upon channel gating, with block developing rapidly on membrane depolarization, but with no preference for activated over inactivated channels. High external [K(+)] (94 mM) reduced the potency of I(HERG) inhibition by both DRONED and AMIOD. Strikingly, mutagenesis to alanine of the S6 residue F656 (F656A) failed to eliminate blockade by both DRONED and AMIOD, whilst Y652A had comparatively little effect on DRONED but some effect on AMIOD. These findings demonstrate that high affinity drug blockade of I(HERG) can occur without a strong dependence on the Y652 and F656 aromatic amino-acid residues.     

Inhibition of the HERG K+ channel by the antifungal drug ketoconazole depends on channel gating and involves the S6 residue F656

The mechanism of human ether-à-go-go-related gene (HERG) K+ channel blockade by the antifungal agent ketoconazole was investigated using patch-clamp recording from mammalian cell lines. Ketoconazole inhibited whole-cell HERG current (IHERG) with a clinically relevant half-maximal inhibitory drug concentration (IC50) value of 1.7 microM. The voltage- and time-dependent characteristics of IHERG blockade by ketoconazole indicated dependence of block on channel gating, ruling out a significant role for closed-state channel inhibition. The S6 HERG mutations Y652A and F656A produced approximately 4-fold and approximately 21-fold increases in IC50 for IHERG blockade, respectively. Thus, ketoconazole accesses the HERG channel pore-cavity on channel gating, and the S6 residue F656 is an important determinant of ketoconazole binding.     

Erythromycin block of the HERG K+ channel: accessibility to F656 and Y652

The HERG potassium channel might have a non-canonical drug binding site, distinct from the channel's inner cavity, that could be responsible for elements of closed-state pharmacological inhibition of the channel. The macrolide antibiotic erythromycin is a drug that may block unconventionally because of its size. Here we used whole-cell patch-clamp recording at 37 degrees C from heterologously expressed HERG channels in a mammalian cell line to show that erythromycin either produces a rapid open-state-dependent HERG channel inhibition, or components of both open-state-dependent and closed-state-dependent inhibition. Alanine-substitution of HERG's canonical determinants of blockade revealed that Y652 was not important as a molecular determinant of blockade, and that mutation of F656 resulted in only weak attenuation of inhibition. In computer models of the channel, erythromycin could make several direct contacts with F656, but not with Y652, in the open-state model, and erythromycin was unable to fit into a closed-state channel model.     

Molecular determinants of voltage-dependent human ether-a-go-go related gene (HERG) K+ channel block

The structural determinants for the voltage-dependent block of ion channels are poorly understood. Here we investigate the voltage-dependent block of wild-type and mutant human ether-a-go-go related gene (HERG) K(+) channels by the antimalarial compound chloroquine. The block of wild-type HERG channels expressed in Xenopus oocytes was enhanced as the membrane potential was progressively depolarized. The IC(50) was 8.4 +/- 0.9 microm when assessed during 4-s voltage clamp pulses to 0 mV. Chloroquine also slowed the apparent rate of HERG deactivation, reflecting the inability of drug-bound channels to close. Mutation to alanine of aromatic residues (Tyr-652 or Phe-656) located in the S6 domain of HERG greatly reduced the potency of channel block by chloroquine (IC(50) > 1 mm at 0 mV). However, mutation of Tyr-652 also altered the voltage dependence of the block. In contrast to wild-type HERG, block of Y652A HERG channels was diminished by progressive membrane depolarization, and complete relief from block was observed at +40 mV. HERG channel block was voltage-independent when the hydroxyl group of Tyr-652 was removed by mutating the residue to Phe. Together these findings indicate a critical role for Tyr-652 in voltage-dependent block of HERG channels. Molecular modeling was used to define energy-minimized dockings of chloroquine to the central cavity of HERG. Our experimental findings and modeling suggest that chloroquine preferentially blocks open HERG channels by cation-pi and pi-stacking interactions with Tyr-652 and Phe-656 of multiple subunits.     

A novel hypothesis for the binding mode of HERG channel blockers

We present a new docking model for HERG channel blockade. Our new model suggests three key interactions such that (1) a protonated nitrogen of the channel blocker forms a hydrogen bond with the carbonyl oxygen of HERG residue T623; (2) an aromatic moiety of the channel blocker makes a pi-pi interaction with the aromatic ring of HERG residue Y652; and (3) a hydrophobic group of the channel blocker forms a hydrophobic interaction with the benzene ring of HERG residue F656. The previous model assumes two interactions such that (1) a protonated nitrogen of the channel blocker forms a cation-pi interaction with the aromatic ring of HERG residue Y652; and (2) a hydrophobic group of the channel blocker forms a hydrophobic interaction with the benzene ring of HERG residue F656. To test these models, we classified 69 known HERG channel blockers into eight binding types based on their plausible binding modes, and further categorized them into two groups based on the number of interactions our model would predict with the HERG channel (two or three). We then compared the pIC(50) value distributions between these two groups. If the old hypothesis is correct, the distributions should not differ between the two groups (i.e., both groups show only two binding interactions). If our novel hypothesis is correct, the distributions should differ between Groups 1 and 2. Consistent with our hypothesis, the two groups differed with regard to pIC(50), and the group having more predicted interactions with the HERG channel had a higher mean pIC(50) value. Although additional work will be required to further validate our hypothesis, this improved understanding of the HERG channel blocker binding mode may help promote the development of in silico predictions methods for identifying potential HERG channel blockers.     

Ranolazine block of human Na v 1.4 sodium channels and paramyotonia congenita mutants

The antianginal drug ranolazine exerts voltage- and use-dependent block (UDB) of several Na+ channel isoforms, including Na(v) 1.4. We hypothesized that ranolazine will similarly inhibit the paramyotonia congenita Na(v) 1.4 gain-of-function mutations, R1448C, R1448H, and R1448P that are associated with repetitive action potential firing. Whole-cell Na+ current (I(Na)) was recorded from HEK293 cells expressing the hNa(v) 1.4 WT or R1448 mutations. At a holding potential (HP) of -140 mV, ranolazine exerted UDB (10 Hz) of WT and R1448 mutations (IC 50 = 59 - 71 μM). The potency for ranolazine UDB increased when the frequency of stimulation was raised to 30 Hz (IC 50 = 20 - 27 uM). When the HP was changed to -70 mV to mimic the resting potential of an injured skeletal muscle fibre, the potency of ranolazine to block I(Na) further increased; values of ranolazine IC 50 for block of WT, R1448C, R1448H, and R1448P were 3.8, 0.9, 6.3, and 0.9 uM, respectively. Ranolazine (30 uM) also caused a hyperpolarizing shift in the voltage-dependence of inactivation of WT and R1448 mutations. The effects of ranolazine (30 uM) to reduce I(Na) were similar (~35% I(Na) inhibition) when different conditioning pulse durations (2-20 msec) were used. Ranolazine (10 μM) suppressed the abnormal I(Na) induced by slow voltage ramps for R1448C channels. In computer simulations, 3 μM ranolazine inhibited the sustained and excessive firing of skeletal muscle action potentials that are characteristic of myotonia. Taken together, the data indicate that ranolazine interacts with the open state and stabilizes the inactivated state(s) of Na(v)1.4 channels, causes voltage- and use-dependent block of I(Na) and suppresses persistent I(Na). These data further suggest that ranolazine might be useful to reduce the sustained action potential firing seen in paramyotonia congenita.     

Inhibition of HERG potassium channel current by the class 1a antiarrhythmic agent disopyramide

The Class 1a antiarrhythmic drug disopyramide (DISO) is associated with 'acquired' prolongation of the QT interval of the electrocardiogram (ECG). This potentially proarrhythmic effect is likely to reflect drug actions on ion channels involved in ventricular action potential repolarisation. In this study, we examined the effects of DISO on potassium channels encoded by HERG, as this K channel type has been implicated in both congenital and acquired long-QT syndromes (LQTS). Chinese hamster ovary cells were transiently transfected with HERG cDNA for subsequent whole cell patch clamp recording. HERG tail currents recorded at -40 mV following test pulses to +30 mV were inhibited in a dose-dependent fashion by DISO concentrations within the clinical range (IC50 = 7.23 +/- 0.72 microM; mean +/- SEM). Experiments with 10 microM DISO indicated that the degree of HERG blockade showed some voltage dependence. Further data obtained using an 'envelope of tails' protocol (pulse potential +40 mV) were consistent with a significant role for open-channel blockade at lower drug concentrations. At higher concentrations it is possible that blockade may have involved drug binding to both resting and open channels. Inhibition of the inactivation-deficient mutant HERG-S631A was comparable to that seen for wild-type HERG. Therefore, channel inactivation was not obligatory for DISO to exert its effect. Native delayed rectifier tail currents from rabbit isolated ventricular myocytes were also inhibited by DISO. We conclude (a) that DISO inhibits HERG encoded potassium channels at clinically relevant concentrations and (b) that this action may constitute the molecular basis for acquired LQTS associated with this drug.     

Molecular and functional characterization of common polymorphisms in HERG (KCNH2) potassium channels

Long QT syndrome (LQTS) is a cardiac repolarization disorder that can lead to arrhythmias and sudden death. Chromosome 7-linked inherited LQTS (LQT2) is caused by mutations in human ether-a-go-go-related gene (HERG; KCNH2), whereas drug-induced LQTS is caused primarily by HERG channel block. Many common polymorphisms are functionally silent and have been traditionally regarded as benign and without physiological consequence. However, the identification of common nonsynonymous single nucleotide polymorphisms (nSNPs; i.e., amino-acid coding variants) with functional phenotypes in the SCN5A Na(+) channel and MiRP1 K(+) channel beta-subunit have challenged this viewpoint. In this report, we test the hypothesis that common missense HERG polymorphisms alter channel physiology. Comprehensive mutational analysis of HERG was performed on genomic DNA derived from a population-based cohort of sudden infant death syndrome and two reference allele cohorts derived from 100 African American and 100 Caucasian individuals. Amino acid-encoding variants were considered common polymorphisms if they were present in at least two of the three study cohorts with an allelic frequency >0.5%. Four nSNPs were identified: K897T, P967L, R1047L, and Q1068R. Wild-type (WT) and polymorphic channels were heterologously expressed in human embryonic kidney cells, and biochemical and voltage-clamp techniques were used to characterize their functional properties. All channel types were processed similarly, but several electrophysiological differences were identified: 1) K897T current density was lower than the other polymorphic channels; 2) K897T channels activated at more negative potentials than WT and R1047L; 3) K897T and Q1068R channels inactivated and recovered from inactivation faster than WT, P967L, and R1047L channels; and 4) K897T channels showed subtle differences compared with WT channels when stimulated with an action potential waveform. In contrast to K897T and Q1068R channels, P967L and R1047L channels were electrophysiologically indistinguishable from WT channels. All HERG channels had similar sensitivity to block by cisapride. Therefore, some HERG polymorphic channels are electrophysiologically different from WT channels.     

川芎嗪对猪冠状动脉平滑肌细胞大电导钙激活钾通道的作用

本工作旨在研究川芎嗪对猪冠状动脉平滑肌细胞钾通道的作用,为阐明其扩张冠状动脉血管的机制提供实验依据。采用膜片钳细胞贴附式和内面向外式记录方式观察川芎嗪对猪冠状动脉平滑肌细胞大电导钙激活钾通道(large-conductance Ca2+- activated potassium channels,BKCa channels)的作用,分别用蛋白激酶A(protein kinase A,PKA)抑制剂H-89和蛋白激酶G (protein kinase G,PKG)抑制剂KT-5823处理细胞,再观察川芎嗪对BKCa通道作用的变化。结果表明在研究的0．73-8．07 mmol/L浓度范围,川芎嗪可以剂量依赖性地激活BKCa通道,使通道的开放概率从(0．01±0．003)增加到(0．03±0．01)-(．21± 0．18)(P<0．01,n=10),使通道平均关闭时间从(732．33±90．67)ms降低到(359．67±41．30)-(2．96±0．52)ms(P<0．01, n=10)。川芎嗪的这种激活作用在浴液游离钙离子浓度接近0 mmol/L时也存在。PKA的特异性抑制剂H-89(3 μmol/L)和 PKG的特异性抑制剂KT-5823(1 μmol/L)对川芎嗪激活BKCa通道的作用无影响。以上结果提示:川芎嗪能直接激活冠状动脉平滑肌BKCa通道,这种作用可能是川芎嗪扩张冠状动脉血管的一种重要机制。