环孢霉素A对心肌细胞结构和胞内钙离子浓度的影响

目的:探讨不同浓度的环孢霉素A(cyclosporine,CsA)在不同时间点对心肌细胞形态及胞内钙离子浓度([Ca2+]i)的影响。方法:分别将1×10-3g/L、1×10-2g/L、2×10-2g/L的CsA与原代培养的Wistar乳鼠(1~3 d)心肌细胞(n=6)共孵育,于2 h、4 h、6 h、8 h和12 h后,采用相差显微镜观察各组心肌细胞的结构及收缩性;应用激光共聚焦显微镜观察心肌细胞内[Ca2+]i的变化。结果:不同浓度的CsA作用2 h后,与对照组相比较,各组心肌细胞的形态及收缩性无明显变化。1×10-3g/L和1×10-2g/L CsA组心肌细胞内的[Ca2+]i与对照组比较无明显差异;但2×10-2g/L CsA组心肌细胞内的[Ca2+]i较其他3组明显增加(P0.01)。作用4 h后,随着CsA浓度的增加,CsA组细胞的收缩性逐渐减弱,胞浆内可见小颗粒逐渐增多;各CsA组心肌细胞内的[Ca2+]i与对照组比较显著增加(P0.05);与1×10-3g/L CsA和对照组相比较,1×10-2g/L和2×10-2g/L CsA组增加明显(P0.05)。作用6 h后,随着CsA浓度的增加,各浓度CsA组与对照组比较,可见细胞内小空泡逐渐增多,细胞的收缩性逐渐减弱。各组心肌细胞内的[Ca2+]i随着CsA浓度的增加逐渐升高,各组组间具有显著差异(P0.05)。作用8 h和12 h后,1×10-3g/L CsA组的心肌细胞出现部分溶解,1×10-2g/L和2×10-2g/L CsA组心肌细胞的损伤严重,出现大面积死亡、溶解。结论:CsA可引起原代培养的心肌细胞损伤,收缩性减弱,且具有剂量和时间依赖性,可能与其引起的细胞内[Ca2+]i的增加有关。     

环孢霉素A肾毒性

１　病例报告１．１　【病例１】　男性患者，３０岁，因肾移植术后１个月，肾功能延迟恢复，于１９９９０２２３入院。缘于１９９０年２月无诱因出现双下肢浮肿，尿检蛋白３＋，隐血＋，未予正规治疗，１９９６年４月发现血压升高达２０／１２ｋＰａ（１５０／９０ｍｍＨｇ），复查血清肌酐（ＳＣｒ）１７６μｍｏｌ／Ｌ。在我院行肾活检确诊为ＩｇＡ肾炎（组织学类型符合局灶性节段性肾小球硬化），给予降压、纠正酸中毒、保肾等综合治疗。１９９８年９月疲劳后头昏、恶心加重伴呕吐，尿量减少，ＳＣｒ升至８００μｍｏｌ／Ｌ以上，诊…     

环孢霉素A治疗原发性肾病综合征

环孢霉素A(Cyclosporin A,CsA)是一种新型的免疫抑制剂,广泛应用于器官移植,抑制免疫排斥反应。1978年Calne等首次用于治疗原发性肾病综合征(PNS),并取得一定疗效. 一、药代动力学 CsA个体药代动力学差异较大,导致临床上个体用药剂量,药效及毒性不同.CsA具有高度脂溶性,有口服及静脉制剂。用药后3～6小时在血中达峰值,主要分布于红细胞及血浆中。CsA半衰期个体差异较大,健康人平均为8.4小时(RIA),肾衰病人为16.1小时(RIA).主要经胆道排泄,仅6%从尿中排出,其中0.1%为药物原型。 Fallath等实验观察发现,肾衰病人对CsA的     

乳鼠心肌细胞培养

目的　鉴于心肌细胞的培养具有简便、定量、重复性好以及不受神经体液因素的影响等特点 ,使得心肌细胞培养有着广阔的应用前景。本文将心肌细胞的分离、纯化、培养及其优点进行一综述     

环孢霉素A对心肌缺血-再灌注损伤的保护机制和治疗进展

在溶栓和介入治疗问世之后,通过早期再灌注治疗使减少心肌梗死面积成为了可能。而再灌注之后的二次心肌损伤使得早期再灌注疗效减弱。目前人们发现环孢霉素A对心肌的保护作用尤为突出,其主要机制是通过抑制线粒体通透性转换孔的开放,从而阻滞了心肌细胞死亡的级联反应。现综述环孢霉素A对心肌缺血-再灌注损伤的保护作用的最新机制、治疗效果和应用前景,旨在为心肌梗死的临床治疗寻找新的更佳途径。     

环孢霉素A治疗小儿难治性肾病中环孢霉素亲和素基因表达的临床意义

目的：研究难治性肾病患儿ＣｓＡ治疗前后环孢霉素亲和素（ＣｙＰ）ｍＲＮＡ的表达及ＣｓＡ的浓度与ＣｙＰｍＲＮＡ基因表达之间的关系，探讨ＣｙＰ在儿童难治性肾病中的临床意义。方法：用ＣｓＡ５ｍｇ／（ｋｇ．ｄ）治疗２６例，其中１６例用逆转录聚合酶链反应（ＲＴ－ＰＣＲ）法测定了治疗前后血白细胞中ＣｙＰｍＲＮＡ的基因表达，并以正常儿童作对照及测定ＣｓＡ血浓度。结果：难治性肾病患儿用ＣｓＡ治疗前血白细胞中ＣｙＰｍＲＮＡ基因表达明显高于治疗后和对照组，而ＣｓＡ血浓度与ＣｙＰｍＲＮＡ基因表达之间成负相关关系。结论：低浓度的ＣｓＡ与高水平表达的ＣｙＰ对肾病不能起到治疗作用，而ＣｓＡ血浓度在１００～２００μｇ／Ｌ之间与低水平表达的ＣｙＰ有助于肾病的缓解。     

环孢霉素A对预防鼠感染疟原虫的实验研究

在小鼠感染伯氏疟原虫的当天，同时经尾静脉注入环孢霉素Ａ（１ｍｇ／ｋｇ·ｄ×２，５ｍｇ／ｋｇ·ｄ×２），观察该药对小鼠感染伯氏疟原虫的预防作用。结果表明：环孢霉素Ａ（５ｍｇ／ｋｇ·ｄ×２）对鼠感染疟原虫有明显的预防效果。提示经尾静脉途径给药环孢霉素Ａ明显比皮下途径给药更具优点。     

葡萄糖浓度变化对乳鼠心肌细胞的影响及其机制的探讨

目的 观察不同浓度的葡萄糖培养对乳鼠心肌细胞形态及功能的影响,并初步探讨其机制.方法 建立乳鼠心肌细胞培养模型,随机分成低糖(5.5 mmol/L)组、高糖(25.5 mmol/L)组、间歇性高糖(5.5 mmol/L和25.5 mmol/L交替培养)组以及高糖(25.5 mmol/L)+蛋白激酶C(PKC)抑制剂Ro-31-8220(50 nmol/L)组,培养5 d后,观察心肌细胞搏动情况,测定心肌细胞直径,并应用Western blot检测心肌细胞PKC-α、PKC-β2、p-PKC-α、p-PKC-β2、核因子(NF)-κB和c-fos的蛋白表达水平.结果 不同浓度葡萄糖培养的心肌细胞在搏动频率、细胞直径以及PKC-α、PKC-β2、p-PKC-α、p-PKC-β2、NF-κB和c-fos的蛋白表达水平存在差异,其中高糖组心肌细胞搏动频率、细胞直径以及PKC-α、PKC-β2、p-PKC-α、p-PKC-β2、NF-κB和c-fos的蛋白表达水平显著高于低糖组[高糖组比低糖组:搏动频率(64.15±1.55)次/min比(57.22±1.91)次/min,细胞直径(20.11±0.91)μm比(17.18±0.82)μm,P均＜0.01],间歇性高糖组的上述指标则显著高于高糖[间歇性高糖组比高糖组:搏动频率(66.60±2.02)次/min比(64.15±1.55)次/min,细胞直径(22.39±0.92)μm比(20.11±0.91)μm,P均＜0.01]和低糖组[间歇性高糖组比低糖组:搏动频率(66.60±2.02)次/min比(57.22±1.91)次/min,细胞直径(22.39±0.92)μm比(17.18±0.82)μm,P均＜0.01],而PKC抑制剂Ro-31-8220则能逆转高糖所诱导的上述变化[高糖+PKC抑制剂Ro-31-8220组比高糖组:搏动频率(62.17±1.58)次/min比(64.15±1.55)次/min,细胞直径(18.41±0.78)μm比(20.11±0.91)μm,P均＜0.01].结论 高糖或间歇性高糖的环境能够导致心肌细胞肥大,搏动频率增加,同时影响心肌细胞PKC-α、PKC-β2、NF-κB和c-fos的表达和活性,而PKC抑制剂Ro-31-8220能够逆转高糖所诱导的上述变化.因此高糖特别是间歇性高糖可能通过PKC/NF-κB/c-fos途径诱导了心肌细胞结构和功能的改变,导致了糖尿病心肌损害.     

吡格列酮对体外培养大鼠破骨细胞样细胞分化及活性的影响

目的 观察吡格列酮对大鼠破骨细胞样细胞(OLC)分化及活性的影响,探讨PPARγ2活化与破骨细胞之间的关系.方法 用NF-кB受体活化因子配体(RANKL)及巨噬细胞集落刺激因子(M-CSF)协同诱导大鼠骨髓单个核细胞(BMC)向OLC分化,同时加入1、5、10μmol/L盐酸吡格列酮干预.应用RT-PCR分析OLC分化过程中PPARγ2及NF-кB受体活化因子(RANK)mRNA的表达,结合细胞染色及抗酒石酸酸性磷酸酶(TRAP)活性观察吡格列酮对OLC分化的影响,通过骨吸收陷窝分析及整合素β3(CD61)平均荧光强度的检测比较吡格列酮对OLC活性的影响.结果 (1)对OLC分化的影响:培养7 d时10μmol/L吡格列酮组与其它组相比OLC数量及TRAP活性明显减少(P<0.01或P<0.05),表明吡格列酮部分抑制OLC分化且此作用与剂量相关;RT-PCR结果显示在诱导分化早期,不同浓度吡格列酮呈剂量依赖性上调PPARγ2的表达水平,但随OLC的成熟其表达渐减弱,而RANK表达在5及10 p,mol/L吡格列酮干预组培养的各时间点均明显下调,与对照及1μmol/L干预组相比有显著差异(P<0.05或P<0.01);(2)对OLC活性的影响:骨吸收陷窝数量及吸收面积在吡格列酮5、10μmol/L组明显减少,与另两组相比差异显著(P<0.05或P<0.01),培养早期这两组细胞CD61平均荧光强度均明显下调(P<0.05或P<0.01).结论 吡格列酮激活PPAR田γ2转录活性可部分抑制大鼠骨髓OLC的分化及活性.     

环孢霉素A对大鼠心肌结构及心脏功能的影响

目的探讨不同剂量环孢霉素A(cyclosporine A,CsA)对大鼠心脏功能的影响以及钙调神经磷酸酶(calcineurin,CaN)活性、mRNA表达与心功能的关系。方法 70只健康纯系Wistar大鼠,体质量(200±25)g, 随机分为A、B、C、D组及正常对照组,每组14只。A、B、C、D组分别给予CsA 5．0、12．5、25．0、50．0 mg·kg-1·-1灌胃,对照组用4 ml生理盐水灌胃。持续给药2、4周后每组各取7只进行心脏超声检测心脏功能、病理形态学、 CaN活性及RT-PCR方法检测(CaNAB mRNA的表达。结果与正常组比较,用药各组随剂量增加心脏功能降低,左室射血分数(LVEF)下降(P<0．01),左室短轴缩短率(FS)下降(P<0．01),A、B、C组左心室(LV)增大,B、 C组左心房(LA)也增大(P<0．05)。随着药物剂量增加,病理改变为心肌变性损伤加重。用药各组2周时(CaN 活性随药物剂量增加而下降(F=27．19,P<0．05),而4周时B、C组CaN活性均较2周时升高(P<0．05); CaNAB mRNA表达随着用药剂量和用药时间增加而下降(P<0．01)。结论大剂量CsA对心肌细胞损伤发生的早而且严重,心脏功能下降明显。     

Rapid, non-perfusion-limited calcium exchange in cultured neonatal myocardial cells

Neonatal rat myocardial cells culture are used to study the effects of zero [Ca]0 and lanthanum (La3+) on contractile function and to correlate these findings with steady-state 45Ca exchange. A rapid superfusion system is used to study cell shortening. At a stimulation rate of 1 Hz exposure to Ca2+ free solution for 900 ms abolishes contraction. Contraction is fully restored within 900 ms after return to 1 mM Ca2+ solution. Similarly, contraction is abolished 800 ms after changing to a 1 mM La3+ solution, but does not return for several min after removal of La3+. A modification of the scintillation-disc flow cell technique is used to study steady-state 45Ca exchange. Cells are grown directly on scintillant plastic discs pre-treated by the Primaria process which enhances cell adhesiveness. The cells on the discs are inserted into a specially designed flow cell which is then perfused at a rate of 500 ml/min. In this manner the labeling period, intervention and washout are performed while instantaneously and continuously monitoring the 45Ca activity without the need to interpose an unmonitored wash period. In addition, the use of on-line computer monitoring allows an accurate and precise recording of data as often as 6 times/s. This permits the identification of the following four Ca2+ compartments: (1) A rapidly exchanging compartment which is La3+ displaceable, contains 2.59 mmols Ca2+/kg dry cells and has a t1/2 = 1.5 s; (2) An intermediate component composed of two compartments which contain 0.43 and 0.22 mmols Ca2+/kg dry cells and exchange with t1/2 of 19 and 103 s respectively; (3) A slowly exchangeable compartment which contains 0.69 mmols Ca2+/kg dry cells with t1/2 = 19 min; (4) An unexchangeable compartment which contains 6.24 mmols Ca2+/kg dry cells. The rapid compartment is La3+ displaceable and has a flux of greater than 1100 mumols Ca2+/kg dry cells/s. This is the first time that the kinetics and sizes of the rapid Ca2+ compartment and intermediate compartments have been measured. Correlation of contractile response to zero [Ca]0 and La3+ administration with 45Ca exchange measurements under conditions of ultra-rapid perfusion indicate that at least a portion of the La3+-displaceable rapid component is critical in the beat-to-beat control of contractile force.     

硒对缺氧培养乳鼠心肌细胞存活率和膜流动性的影响

采用Ｌａａｒｓｅ等培养乳鼠心肌细胞的方法，观察加入不同剂量的亚硒酸钠对“缺氧”心肌细胞成活率及其膜流动性的影响。结果表明：适量硒（１×１０－６ｍｏｌ／Ｌ）可增加细胞存活率及膜流动性，而过量硒（１×１０－５ｍｏｌ／Ｌ）使细胞存活率及膜流动性下降。结论：适量硒对缺氧培养的心肌细胞有保护作用，过量硒具有明显的细胞毒性作用     

重组人脑利钠肽对急性心肌梗死大鼠心功能和心肌细胞凋亡的影响

目的观察重组人脑利钠肽对急性心肌梗死(AMI)大鼠心功能和心肌细胞凋亡的影响。方法通过结扎100只SD大鼠左冠状动脉前降支建立AMI模型,随机分为观察组和对照组各50只。观察组予重组人脑利钠肽15 ug/kg、连续4周,对照组予等体积生理盐水。两组分别于治疗48 h、1周、2周、3周和4周时采用超声检测左室射血分数(LVEF),处死大鼠并取血测定血浆BNP浓度,原位末端标记法(TUNEL法)测定心肌细胞凋亡指数(MAI),免疫组化法检测Fas、FasL蛋白阳性染色指数。结果与对照组比较,观察组LVEF在第48 h即有明显升高(P<0.05);观察组MAI治疗后3、4周与48 h比较下降明显(P均<0.05),观察组治疗4周时与对照组比较亦有明显下降(P<0.05)。观察组于2、3、4周时Fas/Fasl蛋白阳性率明显低于48 h时(P均<0.05),且与对照组比较下降明显(P均<0.05)。结论连续予重组人脑利钠肽能够通过减少大鼠AMI后心肌细胞凋亡,达到改善心功能的目的。     

牛磺酸对大鼠培养心肌细胞钙转运功能的作用

以大鼠培养心肌细胞的脂质过氧化损伤模型，采用４５Ｃａ示踪技术，直接进行心肌肌浆网、线粒体钙转运功能的测定，同时观察牛磺酸对心肌过氧化损伤的保护作用。结果表明，牛磺酸能显著改善心肌肌浆网、线粒体钙转运功能，对心肌细胞具有保护作用     

Effects of exercise training on cardiac function and myocardial remodeling in post myocardial infarction rats

To test the hypothesis that early exercise training after myocardial infarction (MI) could preserve cardiac function, alleviate left ventricular (LV) remodeling and induce a protective effect on morphology, male Sprague-Dawley rats underwent coronary ligation or sham operation, and were assigned to 3 groups: Sham, sedentary MI (SedMI), and exercise MI (ExMI). We measured the changes in collagen volume fraction, matrix metalloproteinase (MMP) 1, tissue inhibitor matrix metalloproteinase 1 (TIMP-1), angiotensin II receptor type 1 (AT1), and angiotensin converting enzyme (ACE) at gene and protein levels after 8 weeks of exercise training. Cardiac functions were determined by echocardiographic and hemodynamic measurements. Early exercise training after MI had no effect on LV wall thinning. Cardiac function was significantly preserved in the ExMI group in comparison to the SedMI group. The collagen volume fraction in the ExMI group was significantly lower than in the SedMI group. Compared to the SedMI group, the ExMI group showed a markedly decrease at both the gene and protein levels in TIMP-1 (P < 0.05). No significant differences were found in MMP-1 among the three groups. MMP-1/TIMP-1 ratio in the ExMI group was significantly higher than in the SedMI group. In addition, the expression of AT1 protein in the ExMI group was significantly lower than in the SedMI group. Furthermore, both ACE mRNA expression and ACE binding in the ExMI group are significantly decreased compared to the SedMI group. Our results suggest that early exercise training after MI reduces TIMP-1 expression, improves the balance between MMPs and TIMPs, and mitigates the expressions of ACE and AT1 receptor. These improvements, in turn, attenuate myocardial fibrosis and preserve post-MI cardiac function.     

环孢素预处理对大鼠心肌缺血-再灌注损伤的保护作用

目的:研究环孢素(CsA)预处理对大鼠心肌缺血-再灌注损伤的保护作用,并探讨其可能机制.方法:36只10周龄SD大鼠随机分为环孢素预处理(CsA)组、缺血-再灌注(IR)组、假手术(Sham)组,每组12只.应用Medlab生物信号采集处理系统连续监测各组左心室收缩末期压(LVESP)和心电图.再灌注结束后采集血液、心...