福莫特罗对离体培养大鼠心肌组织损伤及合用布地奈德对其影响的探讨

In vitro study on the mechanism of myocardial injury induced by Formoterol and the protective effect of Budesonide in rats

目的:观察福莫特罗(FORM)对离体培养SD大鼠心肌组织的损伤作用及合用布地奈德(BUD)的影响,初步探讨FORM致心肌损伤的机制。方法:以0μmol/L、0.01μmol/L、0.05μmol/L、0.1μmol/L、0.5μmol/L、1μmol/L、5μmol/L FORM单独及联合10μmol/L的BUD作用于成年SD大鼠离体心脏组织,并培养72 h。ELISA检测组织培养上清液中肌钙蛋白I(cTnI)和心型脂肪酸结合蛋白(H-FABP)含量,RT-PCR检测心脏组织糖原合成酶激酶-3β(GSK-3β),Western blot检测肌浆网钙泵(SERCA2)、GSK-3β蛋白表达水平。结果:(1)组织培养上清液中,cTnI、H-FABP蛋白随着FORM浓度的升高而增加,除外高浓度的两组,各组间差异有统计学意义(P<0.05)。与单独应用相应浓度的FORM比较,联用BUD后cTnI、H-FABP蛋白浓度降低(P<0.05)。(2)随着FORM浓度的升高,SERCA2蛋白表达降低(P<0.05),GSK-3β蛋白表达增高(P<0.05),但GSK-3βmRNA表达无统计学差异。结论:(1)FORM可致心肌损伤,且呈浓度依赖性,联合应用BUD对FORM所致的心肌损伤有保护作用。(2)FORM导致心肌组织损伤的机制与GSK-3β介导的SERCA2蛋白表达量降低有关。

AIM： To investigate the mechanism of myocardial injury induced by Formoterol（FORM） in vitro and the protective effect of Budesonide（BUD） against such cardiac damage in rats.METHODS： FORM was added in the media of isolated rat myocardial tissues in a concentration gradients of 0 μmol/L,0.01 μmol/L,0.05 μmol/L,0.1 μmol/L,0.5 μmol/L,1 μmol/L,5 μmol/L,respectively,or combined with BUD in a concentration of 10 μmol/L,for 72 hours in vitro.The cardiac troponin I（cTnI） and heart-type fatty acid-binding protein（H-FABP） in supernatant were detected with ELISA method.The mRNA level of glycogen synthase kinase-3β（GSK-3β） was detected with RT-PCR.Sarco（endo） plasmic reticulum Ca2＋-ATPase（SERCA2） and GSK-3β protein expression were detected with Western blot.RESULTS：（1） The values of cTnI and H-FABP were significantly higher in FORM groups,compared with the control group（P〈0.05）,and there was a difference between different FORM concentration gradients（P〈0.05） except 1 μmol/L and 5 μmol/L groups.In FORM and BUD combined groups,the values of cTnI and H-FABP were significantly lower than FORM groups（P〈0.05）.（2） With the increase of FORM concentration,there were lower expression of SERCA2（P〈0.05） and higher expression of GSK-3β protein（P〈0.05）.But there was no significant statistic difference of GSK-3β mRNA between FORM treated groups（P〈0.05）.CONCLUSION：（1） FORM can induce injury in rat＇s isolated myocardial tissues and the injury is concentration depended.BUD shows protective effect against FORM induced myocardial injury.（2） The mechanism of FORM induced myocardial injury may be associated with the lower expression of SERCA2,which is mediated by GSK-3β.