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目的:比较苦参碱、青蒿素和粉防己碱对分离的豚鼠心室肌细胞胞浆钙的影响。方法:用酶解法急性分离的豚鼠心室肌细胞先用Fluo 3-AM负载,然后用激光扫描共聚焦法检测单个豚鼠心室肌细胞胞浆钙的荧光强度。结果:(1)KCl 60mmol·L~(-1)可升高心室肌细胞胞浆钙荧光强度,其荧光强度从299±19升高到1389±325(P<0.01)。(2)苦参碱和青蒿素在浓度为100μmol·L~(-1)可使KCl 60mmol·L~(-1)引起的外钙内流增多,其荧光强度分别从301±14和299±16升高到1495±320和1646±308(P<0.05)。(3)粉防己碱1、10及100μmol·L~(-1)和维拉帕米10μmol·L~(-1)可抑制KCl引起的外钙内流。(4)在正常细胞外液中,苦参碱1、10和100μmol·L~(-1)可使细胞内荧光强度增强,其荧光强度分别从303±27,300±32,296±19上升到441±96,504±112,643±126(P<0.05)。(5)在无钙细胞外液中,粉防己碱1和10μmol·L~(-1)可明显抑制咖啡因20mmol·L~(-1)引起的胞浆钙升高(P<0.05)。结论:苦参碱、青蒿素和粉防己碱对豚鼠心室肌细胞胞浆钙的影响不同。青蒿素和苦参碱可加强电压依赖性钙内流,而粉防己碱与维拉帕米作用相似,抑制这种钙内流。苦参碱本身可以促进外钙内流。
OBJECTIVE: To compare the effects of matrine, artemisinin and tetrandrine on cytosolic calcium in isolated guinea pig ventricular myocytes. METHODS: Acute isolated guinea pig ventricular myocytes were enzymatically hydrolyzed with Fluo 3-AM. Laser fluorescence confocal microscopy was used to detect the cytosolic calcium fluorescence intensity of guinea pig ventricular myocytes. Results: (1) KCl 60 mmol·L -1 increased the cytoplasmic calcium fluorescence intensity of ventricular myocytes, and the fluorescence intensity increased from 299±19 to 1389±325 (P<0.01). (2) The extracellular calcium influx induced by KCl 60 mmol·L -1 increased with matrine and artemisinin at concentrations of 100 μmol·L -1 , and their fluorescence intensities were from 301±14 and 299, respectively. ±16 increased to 1495±320 and 1646±308 (P<0.05). (3) Trindrine 1, 10, and 100 μmol·L -1 and verapamil 10 μmol·L -1 inhibited the extracellular calcium influx induced by KCl. (4) In normal extracellular fluid, matrine 1, 10, and 100 μmol·L -1 enhanced intracellular fluorescence intensity, and their fluorescence intensity increased from 303±27,300±32,296±19 to 441±96,504 respectively. ± 112,643 ± 126 (P < 0.05). (5) In the calcium-free extracellular fluid, tetrandrine 1 and 10 μmol·L -1 could significantly inhibit the increase of cytosolic calcium induced by caffeine 20 mmol·L -1 (P<0.05). CONCLUSIONS: Matrine, artemisinin, and tetrandrine have different effects on cytosolic calcium in guinea pig ventricular myocytes. Artemisinin and matrine enhance voltage-dependent calcium influx, whereas tetrandrine and verapamil act similarly and inhibit this calcium influx. Matrine itself can promote extracellular calcium influx.