心肌缺血再灌注损伤大鼠胱硫醚β-合酶／硫化氢和血红素氧合酶-1／一氧化碳的相互作用

Interaction between endogenous cystathionine synthase/hydrogen sulfide and heme oxygenase-1/ carbon monoxide systems during myocardial ischemic-reperfusion： experiment with rats

目的研究胱硫醚β-合酶（CBS）／硫化氢（H2S）体系和血红素氧合酶-1（HO-1）／一氧化碳（CO）体系在大鼠心肌缺血再灌注损伤中的相互作用。方法30只SD大鼠随机分为5组（n=6）：对照组（C组）、心肌缺血再灌注组（I／R组）、心肌缺血再灌注＋锌原卟啉（HO-1抑制剂）组（I／R＋Z组）、心肌缺血再灌注＋羟氨（CBS抑制剂）组（I／R＋H组）、心肌缺血再灌注＋锌原卟啉＋羟氨组（I／R＋Z＋H组）。I／R＋Z组、I／R＋H组和I／R＋Z＋H组夹闭左冠状动脉前30min分别腹腔注射锌原卟啉45μmol／kg、5mmol／L羟氨1ml和5mmol／L羟氨1ml＋锌原卟啉45μmol／kg，C组和I／R组腹腔注射等量生理盐水。缺血30min再灌注2h后处死大鼠取心肌组织，测定大鼠心肌组织中H2S、NO、GSH、MDA含量、SOD活性及CBS-mRNA和HO-1-mRNA表达的变化；电镜下观察心肌组织线粒体的变化。结果与C组相比，I／R组H2S、CO和MDA含量增高，GSH含量和SOD活性降低，CBS-mRNA和HO-1-mRNA表达增高。与I／R组相比，I／R＋Z组CO含量降低，H2S和GSH含量增高，CBS-mRNA表达增高，HO-1-mRNA表达降低；I／R＋H组H2S和GSH含量降低，CO含量增高，CBS-mRNA的表达降低，HO-1-mRNA表达增高；I／R＋Z＋H组H2S、CO、GSH含量和SOD活性降低，MDA含量增高，CBS—mRNA和HO-1-mRNA的表达均降低；I／R＋Z＋H组大鼠心肌组织线粒体损伤加重（P〈0．05）。结论CBS／H2S体系和HO-1／CO体系在大鼠心肌缺血再灌注损伤过程中有相互作用。

Objective To investigate the interaction between the cystathionine synthase gene （CBS）/ hydrogen sulfide （H2S） and heine oxygenase-1 （HO-1）/carbon oxide （CO） systems during myocardial ischemia-reperfusion. Methods Twenty-four SD rats were randomly divided into 4 equal groups： ischemia-reperfusion （I/R） group undergoing intraperitoneal injection of normal saline （NS） and then 30 mix later undergoing ligation of the left coronary artery （LCA） for 30 mix, isehemia-repeffusion ＋ zinc protoporphyrin （I/R ＋ Z） group undergoing intraperitoneal injection of 45 μmoL/kg zinc protoporphyrin, a HO-1 inhibitor, 30 mix before the ligation of LCA, ischemia-reperfusion ＋ hydroxylamine （I/R ＋ H） group, undergoing intraperitoneal injection of 5 mmoL/L hydroxylamine, a CBS inhibitor, 30 mix before the ligation of LCA, and ischemia-reperfusion ＋ zinc protoporphyrin ＋ hydroxylamine （ I/R ＋ Z ＋ H ） group undergoing intraperitoneal injection of zinc protoporphyrin and hydroxylamine 30 mix before the ligation of LCA. Another 6 rats underwent sham operation and intraperitoneal injection of NS 30 mix before the operation and were used as control （C） group. The rats were killed 2 h after the reperfusion with their hearts taken out. Electron microscope was used to observe the mitoehondrial structure. Homogenate of myocardium was prepared to examine the concentrations of H2S, CO, glutathione （GSH）, malonyldialdehyde （MDA）, activity of superoxide dismutase （ SOD）, and expression level of CBS-mRNA and HO-I-mRNA. Result The concentration of CO, H2S, GSH, and MDA increased, SOD activity decreased, and HO-I-mRNA and CBS-mRNA expression levels increased in Group I/R as compared with Group C （ all P 〈 0.01 ）. In Group I/R ＋ Z the CO level decreased, H2S and GSH levels increased, and HO-I-mRNA expression level decreased, but CBS-mRNA expression level increased as compared with Group I/R （ all P 〈 0.05 ）. In Group I/R ＋ H the H2S and GSH levels decreased, CO level increased, and HO-I-mRNA expression level increased and CBS-mRNA expression level decreased as compared with Group I/R （ all P 〈 0.05 ）. In Group I/R ＋ Z ＋ H the H2 S, CO, GSH, and SOD levels decreased, MDA level increased, and HO-I-mRNA and CBS-mRNA expression levels decreased as compared with Group I/R （ all P 〈 0. 05 ）. Electron microscopy found that mitochondrial structure was destroyed in the Groups I/R, I/R ＋ Z, I/R ＋ H, and most seriously in Group I/R ＋ Z ＋ H. Conclusion Both CBS/H2S and HO-1/CO systems play a protective role in myocardial ischemia-reperfusion and they interact on each other.