Abnormal calcium transport ofmyocardial mitochondria during the early stage of combined radiation－burn injury in rats

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Effect of Calcium Ionophore and Calcium Channel Blockers on Immediate Hypersensitivity Reactions

The release of mediators from mast cells is a model for cell secretion and is an in-vitro index for immediate hypersensitivity reactions. Calcium influx is generally accepted to be the primary biochemicalevent in mast cell activation.We studied the effect of the calcium ionophore A 23187 and calcium channelblockers,nifedipine and verapamil, in triggering the activation of rat peritoneal mast cells.At suitableconcentration nifedipine and verapamil have had the inhibition effect in the IgE-dependent roaction.

Experimental study of verapamil on the relaxation of isolated human corpus cavernosum tissues

Aim： To evaluate the relaxant effect of verapamil on human corpus cavernosum in vitro and to assess the drug＇s potential as a treatment for erectile dysfunction （ED）. Methods： Preparations of the human corpus cavernosum were obtained from recently deceased young men who had had normal erectile function. The isometric tension and detailed curves were recorded when contractions induced by 10 mmol/L phenylephrine were reduced by different doses of verapamil or the vehicle control （sterile water）. The tension of human corpus cavernosum preparations are described as a percentage of their top tension before adding verapamil or the vehicle. ANOVA and least significant difference tests were used for statistical analysis. Results： Doses of 1μmol/L, 10 μmol/L and 100 μmol/L verapamil resulted in relaxation of （35.28 ± 7.96）%, （55.91 ± 6.41）%, （85.68 ± 4.16）% after 30 min, respectively. The vehicle control at the same time point produced relaxation of （-0.06 ± 10.57）% （P 〈 0.05）. Conclusion： Verapamil is significantly effective in relaxing normal human corpus cavernous smooth muscle induced by phenylephrine in vitro and the relaxant effect depends on the concentration of verapamil. （Asian J Androl 2006 Mar; 8： 195-198）

Role of Calcium in Glucagon Secretion

The role of calcium ions in the process by which nutrients affect glucagon secretion by pancreatic islet α-cells remains the matter of an apparently endless debate. In the prolongation of recent articles dealing with this matter, the present review draws attention to the dual role of Ca2+ as revealed by prior publications. In such a perspective, emphasis is placed on the increase in glucagon output in response to the omission of extracellular Ca2+ as recorded in the presence of D-glucose or 2-ketoisocaproate, the permissive role of extracellular Ca2+ in the positive secretory response to arginine or a mixture of fumarate, glutamate and pyruvate, and the effects of an organic calcium-antagonist on glucagon output. Considering the role currently ascribed to Ca2+ in the activation of motile events involved in stimulus-secretion coupling, attention is also given to the effects of cytochalasin B, D2O and mitotic-spindle inhibitors upon the secretory response of α-cells exposed to D-glucose in the absence or presence of arginine.

Verapamil modulating arsenic trioxide-induced QT interval prolongation in guinea pig

Objective To investigate therapeutic action of verapamil on QT prolongation induced by arsenic trioxide (As2O3) in guinea pig and to further explore its possible mechanism. Methods Different doses of As2O3 was infused intravenously to observe the changes of QT interval on the electrocardiogram (ECG) at different times in guinea pig.Patch clamp technique and laser scanning confocal microscopy were utilized to study the action of As2O3 on action potential duration (APD),L-type calcium current (ICa-L), rapid delayed rectifier potassium current (IKr) and intracellular calcium concentration ([Ca2+]i) of guinea pig myocytes. At the same time, verapamil was applied preliminarily to evaluate effects of verapamil on changes of the above index induced by AS2O3 . Results Intravenous administration of As2O3 at the dose of 1.6mg/kg and 0.8mg/kg prolonged QT interval on ECG obviously in guinea pig hearts dose dependently and time de pendently. QTc (corrected QT interval) was progressively prolonged in the 2-hour period of intravenous infusion of 1.6mg/ kg As2O3 from (328 .5±30.9)ms of control to (388 .4±31. 3)ms at 2h following As2O3 (P < 0.01). When verapamil was pretreated for 5min,then 1.6mg/kg As2O3 was added,the results showed that QTc was shorter in verapamil-treatment group (357 .3±21 .4)ms than that in As2O3 group (388 .4±31.3)ms (P<0.05) at 2h.Confocal experiments showed that in normal Tyrode solution, As2O3 (1μmol/L and 10/μmol/L) had no obvious effects on resting [Ca2+ ]i( P > 0 .05) in guinea pig cardiomyoeytes,however, 10μmol/L As2O3 could markedly enhance [Ca2+ ]. increase induced by KCl 60mmol/ L and the peak value increased from 903 .4±369.4 to 1674. 6±563 .2 ( P < 0.05 ) . The action of elevating [ Ca2+ ]i could be blocked by 10μmol/L verapamil incompletely. The patch-clamp studies indicated that As2O3 at concentration of 10μmol/L prolonged APD50 from (263 .6±75 .2)ms to (523.9±47 .8)ms (P<0.01) and APD90 from (277.5±77.5) ms to (536.3±49.6)ms (P<0.01),and increased ICa-L from ( -6.0±1.5)pA/pF to ( -8.7±2.0)pA/pF (P < 0.01) at 0mV and also reduced IKr from (6.7±1.8)pA/pF to (4.5±1 .8)pA/pF ( P < 0.05) .However,10μmol/L verapamil could modulate prolonging APD50 from (523 . 9±47 . 8 ) ms to (340.4±83 . 8 ) ms ( P < 0.01) and APD90 from (536.3±49.6) ms to (348.9±85.5)ms (P < 0.01) and correct increasing ICa-L induced by 10μmol/L As2O3 from ( - 8 .7±2.0) pA/pF to ( - 6.6±1.4) pA/pF ( P < 0.05) at 0mV. Conclusion As2O3 could induce prolongation of the QT interval on the ECG in guinea pig hearts and the ionic mechanism is associated with increasing ICa-L and inhibiting IKr/HERG. Verapamil may be useful in normalizing QT prolongation during As2O3 therapy by decreasing ICa-L and [Ca2+]i of ventricular myocytes in guinea pig.

Verapamil inhibits 3T3-L1 preadipocyte differentiation

Objective： To investigate the effect of the calcium channel blocker verapamil on adipocyte differentiation and its mechanism of action. Methods： Preadipocytes from 3T3-L1 strain mouse embryos were cultured and differentiated into matured adipocytes in vitro. Verapamil was added to the culture medium in the concentration of 30 μmol/L on Day 0. Cell differentiation was determined by Oil Red O staining and marker gene mRNA expression was evaluated and compared by RT-PCR. The fluo-3/AM probe and laser scanning confocal microscopy were used to measure intracetlular calcium concentrations. Results： （1）The differentiation rate of 3T3-L1 preadipocytes exposed to verapamil was lower than that of untreated cells. （2）Verapamil promoted the retention of pref-1 gene expression. Lipoprotein lipase expression in the verapamil group was significantly lower than that in the control group on Day 4, Day 6 and Day 8 （P 〈 0.05） and resistin expression was significantly lower than that in the control group on Day 6, Day 8 and Day 10 （P 〈 0.05）. Fatty acid synthase expression in the verapamil group was significantly lower than that in the control group from Day 2 （P 〈 0.05）. （3） Intracellular concentrations of calcium [Ca^2＋]i in the verapamil group were significantly decreased compared with those in the control group on Day 2, Day 4 and Day 6 （P 〈 0.05）, while there was no obvious difference between the two groups on Day 0 （P 〉 0.05）. Conclusion： In 3T3-L1 preadipocytes verapamil significantly reduced adipocyte differentiation, down-regulated the mRNA expression of three marker genes for adipocytes differentiation, and prolonged the mRNA expression of an inhibitor of differentiation. The inhibitory effect of verapamil on differentiation may involve its role as a blocker of calcium influx in adipocytes.

In vitro-inhibition of shear-induced platelet aggregation by verapamil

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Na^(+)/Ca^(2+)交换体抑制剂SN-6和维拉帕米降低肾上腺皮质癌细胞系NCI-H295R醛固酮合成酶表达

目的研究Na^(+)/Ca^(2+)交换体(NCX)抑制剂SN-6和钙通道阻滞剂(CCB)维拉帕米对钾离子(K^(+))刺激的肾上腺皮质癌细胞系NCI-H295R(H295R)醛固酮合成酶表达的影响。方法H295R细胞分为对照组、K^(+)(15 mmol/L)处理组、钙通道阻滞剂维拉帕米(verapamil)(10μmol/L)处理组、SN-6(10μmol/L)处理组、K^(+)+维拉帕米处理组、K^(+)+SN-6处理组、维拉帕米+SN-6处理组和K^(+)+维拉帕米+SN-6处理组,用实时荧光定量PCR检测醛固酮合成酶(CYP11B2)的mRNA表达,用FLIPR Calcium6检测细胞内钙离子水平。结果与对照组相比,K^(+)刺激醛固酮合成酶CYP11B2的mRNA表达(P<0.001);SN-6和维拉帕米均抑制K^(+)刺激的CYP11B2的mRNA表达(P<0.01);与K^(+)+SN-6组相比,K^(+)+SN-6+维拉帕米组更能显著抑制CYP11B2的mRNA表达(P<0.001)。SN-6和维拉帕米显著降低K^(+)刺激的细胞内钙离子水平(P<0.0001)。结论SN-6和维拉帕米均抑制K^(+)诱导的H295R细胞的醛固酮合成酶的表达;SN-6联合维拉帕米处理,抑制作用更显著。

粉防己碱对大鼠心肌细胞电压依赖性钙通道的作用

运用钙离子荧光指示剂Ｆｕｒａ－２／ＡＭ，检测了粉防己碱（Ｔｅｔ）对成年大鼠心室肌细胞电压依赖性钙通道的影响。结果显示：基础状态下心肌细胞内钙离子（［Ｃａ２＋］ｉ）为１６２．６±７．３ｎｍｏｌ·Ｌ－１，５０ｍｍｏｌ·Ｌ－１氯化钾能使［Ｃａ２＋］ｉ增加至４８０．８±９．３ｎｍｏｌ·Ｌ－１（Ｐ＜０．０１），在无细胞外钙条件下，这种增加作用消失，而预先给予Ｔｅｔ和维拉帕米（Ｖｅｒ）则能阻断高钾升高［Ｃａ２＋］ｉ的作用。结果提示：Ｔｅｔ是通过阻断电压依赖性钙通道而发挥作用的。

脱氢紫堇碱对正常和低氧豚鼠心肌细胞内钙的影响

目的 :探讨脱氢紫堇碱 (dehydeocorydaline,DHC)及维拉帕米 (verapamil,Ver)对豚鼠心肌细胞内游离钙浓度([Ca2 + ] i)变化的影响。方法 :采用离体豚鼠心脏Langendorff法灌注 ,用荧光指示剂方法 (Fure 2 /AM)标记心肌([Ca2 + ] i)变化。观察低氧后心肌 [Ca2 + ] i 的变化。结果 :①正常氧状态心肌 [Ca2 + ] i 均值为 (1 2 0 .5± 8.3)nmol/L(n =2 0 ) ;②正常氧条件下 ,DHC、Ver均使心肌 [Ca2 + ] i 明显下降。 (3)低氧状态下 ,心肌 [Ca2 + ] i 增加与缺氧时间(程度 )直线相关 (r=0 .98)。④DHC对低氧后心肌 [Ca2 + ] i 增加明显减缓。结论 :DHC在正常氧、低氧条件下阻止心肌细胞内钙超载 。

大鼠胰腺缺血/再灌注损伤的实验研究

目的 观察大鼠胰腺缺血／再灌注损伤对胰腺组织氧自由基（ＭＤＡ）、胰腺组织钙、胰腺细胞凋亡等的变化及异搏定对缺血／ 再灌注损伤胰腺的保护作用。方法 采用钳闭大鼠腹腔干、肠系膜上动脉制备胰腺缺血／再灌注损伤模型。取Ｗｉｓｔａｒ 大鼠６３ 只，随机分为假手术组、缺血／再灌注组、缺血／ 再灌注异搏定保护组。结果 缺血／ 再灌注异搏定保护组大鼠胰腺组织ＭＤＡ、组织钙、胰腺细胞凋亡明显较缺血／ 再灌注组低，病理改变轻。缺血／ 再灌注组胰腺细胞凋亡与ＭＤＡ、组织钙存在相关性。结论 异搏定降低胰腺组织ＭＤＡ和组织钙含量，抑制细胞凋亡，保护缺血／ 再灌注损伤的胰腺；胰腺缺血／ 再灌注损伤中存在细胞凋亡机制的参与；细胞凋亡存在ＭＤＡ和组织钙超载的参与。

戊脉安等钙拮抗剂对平阳霉素细胞毒性的生化调节作用

本文在体外培养的L_(1210)细胞中观察了平阳霉素单用或与钙拮抗剂合用时的细胞毒作用,结果表明:戊脉安本身在无细胞毒性浓度下与平阳霉素联合应用时,可显著加强平阳霉素(75μg/ml)的细胞毒作用。而且这种增效作用随戊脉安浓度的增加而加强,细胞存活率可由平阳霉素单药时的25％,分别降低为0.11％-9.9％。同样,当戊脉安浓度固定为20μg/ml时,增效作用随着平阳霉素浓度的增加而加强。此外,还发现钙调蛋白阻滞剂利多卡因、氯丙嗪及普鲁卡因,对平阳霉素也具有类似的增效作用,其增效幅度也随着上述各种钙阻滞剂的浓度增加而加强。

人参皂甙Rg1对缺氧豚鼠心肌细胞游离钙浓度降低作用的研究

目的 :探讨人参皂甙 (ginsenosides,Gin)单体Rgl及维拉帕米 (verapamil,Ver)对豚鼠心肌细胞内游离钙浓度 ([Ca2 + ]i)变化的影响。方法 :采用离体豚鼠心脏Langendorff法灌注 ,胶原酶I型分离心肌细胞 ,用荧光指示剂方法 (Fura 2 /AM)标记心肌 ([Ca2 + ]i)变化。将心肌细胞悬液分为 3组 :对照组、Rgl组和Ver组。观察缺氧后心肌[Ca2 + ]i 的变化。结果 :(1)正常氧状态心肌 [Ca2 + ]i 均值为 (12 5 .4± 10 .3)nmol/L(n =2 0 )。 (2 )缺氧状态下 ,心肌[Ca2 + ]i 增加与缺氧时间 (程度 )直线相关 ,相关系数r为 0 .98左右。 (3)Rgl对缺氧后心肌 [Ca2 + ]i 增加明显延缓。结论 :Rgl在缺氧条件下 ,使心肌 [Ca2 + ]i 明显下降 ,从而阻止心肌细胞内钙超载 ,其作用与Ver相似 。

汉防己甲素、维拉帕米治疗肺间质纤维化的实验研究

目的:观察中西药钙拮抗剂对肺间质纤维化的治疗作用。方法:将72只 SD大鼠一次性气管内注入博莱霉素 A_5复制肺纤维化模型,次日随机分组给药,并测定分析各组给药后第7、14、28天肺组织内羟脯氨酸、脂质过氧化物含量、组织病理学变化以及比较各组第28天肺系数。结果:汉防己甲素组及维拉帕米组各期肺组织脂质过氧化物及羟脯氨酸含量均较模型组明显降低(P<0.05或P<0.01),肺系数明显减小(P<0.001或 P<0.01),组织病理学显示肺泡炎及肺纤维化程度均明显减轻,其中汉防己甲素优于维拉帕米。结论:中药钙拮抗剂汉防己甲素治疗肺间质纤维化疗效满意,值得进一步研究。

急性胰腺炎患者血浆磷脂酶A_2的变化及维拉帕米的治疗效应

目的观察急性胰腺炎(acute pancreatitis,AP)患者血浆磷脂酶A2(phospholipase A2,PLA2)的变化,探讨其在胰腺和器官功能损伤中的作用及维拉帕米(verapamil)的治疗效应。方法符合AP诊断患者48例,随机分成维拉帕米治疗组和常规治疗组,分别在治疗前(入院时)、治疗后第2、4天清晨空腹抽肘静脉血4mL,测定血浆PLA2活性、血清淀粉酶和血钙含量,并设对照组。结果AP患者血清淀粉酶活性显著高于对照组(P<0.01),维拉帕米治疗组血清淀粉酶活性明显低于常规治疗组(P<0.05或P<0.01);AP患者血浆PLA2活性显著高于对照组(P<0.01),维拉帕米治疗组血浆PLA2活性明显低于常规治疗组(P<0.05或P<0.01);AP患者血钙浓度明显降低,与对照组比较差异有统计学意义(P<0.01),维拉帕米治疗组血钙浓度明显高于常规治疗组(P<0.05或P<0.01);维拉帕米治疗组发生多器官功能不全病例明显少于常规治疗组(P<0.01)。结论PLA2在AP及并发器官功能损伤的病理过程中发挥重要的作用,维拉帕米可以通过钙通道拮抗作用抑制PLA2的活性,减轻胰腺及器官功能的损伤而发挥治疗作用。

川芎嗪抗再灌注心律失常作用的研究

本文旨在探讨川芎嗪（TMP）抗离体大鼠心脏缺氧／复氧再灌注心律失常的作用及机制。结果表明氧反常组在再灌初期均发生心律失常，持续至2．8±2．2min；川芎嗪组心律失常发生率仅为28％．平均持续时间为3．5±1．5min．两组有显著性差异（p<0．01）。其作用机理可能与TMP清除自由基，抗脂质过氧化而维持细胞正常电活动和阻断Ca2+内流有关。研究提示TMP是优于异搏定的Ca2+通道阻断剂。

汉防已甲素治疗博莱霉素诱导的大鼠肺间质纤维化实验研究

目的 观察中西药钙拮抗剂对肺间质纤维化的治疗作用。方法 将 72只SD大鼠一次性气管内注入博莱霉素A5制备肺纤维化模型 ,次日随机给药 ,并测定分析各组给后d7、d14、d2 8肺组织内羟脯氨酸、脂质过氧化物含量、组织病理学变化以及比较各组d2 8肺系数。结果 汉防己甲素组及维拉帕米组各期肺组织脂质过氧化物及羟脯氨酸含量均较模型组明显降低 (p <0 .0 5 ,p<0 .0 1) ,肺系数明显减小 (p <0 .0 1,p<0 .0 1) ,组织病理学显示肺泡炎及肺纤维化程度均明显减轻 ,其中汉防己甲素优于维拉帕米。结论 中药钙拮抗剂汉防己甲素治疗肺间质纤维化疗效满意 ,值得进一步研究。

盐酸丙哌维林对豚鼠离体膀胱平滑肌条的钙拮抗作用

目的 研究盐酸丙哌维林 (P 4)对豚鼠离体膀胱平滑肌条的钙拮抗作用及作用机制。方法 采用离体平滑肌条浴槽实验方法 ,以维拉帕米为对照 ,测定P 4对CaCl2 和组胺 (His)所致离体膀胱平滑肌条的收缩及对乙酰胆碱 (ACh)所致膀胱平滑肌依赖细胞内钙、外钙收缩的影响。结果 P 41～ 1 0 0 μmol·L- 1 使CaCl2 累积量效曲线非平行右移 ,最大反应降低 ,pD2 ′为 4 73± 0 1 4。P 41 μmol·L- 1 使His量效曲线平行右移 ,最大效应不变 ,pA2 为 5 44± 0 1 4 ;1 0～ 1 0 0μmol·L- 1 使His量效曲线非平行右移 ,最大效应降低 ,pD2 ′为 4 71± 0 1 0。P 41、1 0和 1 0 0 μmol·L- 1 对ACh所致依赖细胞内钙收缩的抑制率分别为 45 77%± 6 54 %、86 2 6 %± 5 59%和 90 55 %± 3 1 1 % ,对依赖细胞外钙收缩的抑制率分别为 7 30 %± 2 89%、49 1 6 %± 6 0 9%和 88 2 5 %±3 70 %。结论 P 4低浓度时竞争性拮抗His所致膀胱平滑肌条的收缩反应 ,明显抑制膀胱平滑肌条依赖细胞内钙释放的收缩反应 ;高浓度时非竞争性拮抗His和CaCl2 所致膀胱平滑肌条的收缩反应 。

维拉帕米增强曲马多镇痛作用的实验研究

目的本实验旨在观察钙通道拮抗剂与曲马多联合外周应用对大鼠的镇痛效应。方法痛阈测定选用电刺激甩尾和热板法。腹腔注射不同剂量的维拉帕米（５，１０，２０ｍｇ·ｋｇ－１）、曲马多（１０，２０，４０ｍｇ·ｋｇ－１）或小剂量维拉帕米（５ｍｇ·ｋｇ－１）和不同剂量曲马多（１０，２０，４０ｍｇ·ｋｇ－１）联合注射。结果曲马多单独应用时，镇痛效应确切，在所试剂量范围内与剂量正相关。维拉帕米单用时，无论剂量大小均无镇痛作用，但小剂量（５ｍｇ·ｋｇ－１）与不同剂量曲马多联用，可显著增强曲马多的镇痛效应，缩短起效时间，延长其有效镇痛期。结论钙通道拮抗剂可增强曲马多的镇痛作用，此联合用药方法简单、方便、危险性小，有在临床推广应用的前景。

龙晶一号对离体大鼠胸主动脉环的作用

龙晶一号(L01)是从飞龙掌血中分离出的有效成分。L01 45～405μmol/L可拮抗KCl和NE所致大鼠主动脉环收缩,其IC_(50)分别为206.93和94.18μmol/L。L01可使CaCl_2的量效反应曲线平行右移,其对PDC的阻滞作用与Ver相似。L01 80μmol/L抑制NE所致细胞内Ca^(2+)释放和外Ca^(2+)内流,对前者作用较显著,与Ver有别。提示L01可能为一种新的钙拮抗剂。