硫化氢对截肢创伤后大鼠肺线粒体损伤的保护作用

Protective effect of hydrogen sulfide on lung mitochondrion function after amputated extremity trauma

目的观察大鼠下肢截肢创伤后肺线粒体的损伤情况及硫化氢(H2S)对截肢创伤应激后肺线粒体功能的保护作用。方法 32只雄性SD大鼠分为对照组、创伤组、硫氢化钠(NaHS)组(截肢后立即腹腔注射NaHS 28μmol/kg)和炔丙基甘氨酸(PPG)组(截肢后立即给予腹腔注射PPG 50 mg/kg),每组8只。通过手术制备大鼠左后肢截肢模型,截肢后6 h处死动物,迅速提取肺组织线粒体,测定肺线粒体呼吸控制率、磷氧比(P/O)、膜电位及三磷酸腺苷(ATP)酶活性,电镜下观察肺线粒体的形态改变。结果与对照组比较,创伤组肺线粒体的呼吸功能受损[呼吸控制率(RCR)、P/O、膜电位及总ATP酶的含量下降(P<0.05)];与创伤组比较,NaHS组肺线粒体RCR、P/O、膜电位及总ATP酶的含量明显升高(P<0.05);PPG组P/O、总ATP酶的含量降低(P<0.05)]。电镜观察显示,创伤组、PPG组肺线粒体肿胀、嵴断裂;而NaHS组线粒体仅轻度肿胀。结论截肢创伤应激可使肺线粒体的呼吸功能受损、膜电位下降、能量代谢障碍;硫化氢对截肢创伤后肺线粒体的损伤具有保护作用。

Objective To investigate the influence of hydrogen sulfide on lung mitochondrion function after amputated extremity trauma and probe the injury of mitochondrion and the protection of H2S on lung mitochondrion function after amputated extremity trauma. Methods 32 aduh male SD rats were randomly divided into 4 groups （ 8 each） ： the control group, the trauma group, NariS group ： NariS solution intraperitoneal injection （ 28 μmol/kg）, and PPG group：PPG solution intraperitoneal injection （50mg/kg）. The rats were sacrificed 6 hours after trauma,and the mitochondria were extracted from lung. The mitochondrial respiratory control ratio （RCR）, P/O, membrane potentials and total ATPase activity were detected. The morphological changes in lung mitochondria were observed by electron microscopy. Results Compared with that in the control group,the respiratory function of mitochondrion was damaged after trauma. RCR, P/O, the membrane potential and the content of ATP in mitochondrion decreased significantly （P 〈 0.05 ）. Compared with that in the trauma group, RCR, P/O, the membrane potential and the content of ATP in mitochondrion increased（ P 〈 0.05 ） in NariS group, the P/O and the content of ATP in mitochondrion were decreased in PPG group （ P 〈 0.05 ）. Ultrastructural examination revealed swelling and disorganization of cristae in mitochondria in traumatic group and PPG group, and only slight swelling of mitochondria was found in NariS group. Conclusions The stress of amputation trauma may result in damage of lung mitochondria, and H2 S seems to be able to attenuate the damage.