他克莫司对截肢创伤后大鼠肝、肾组织线粒体功能的保护及其机制

Protection and mechanism of tacrolimus to the injury of mitochondria in liver and kidney of rats suffered amputation

目的研究截肢创伤后大鼠肝、肾等远隔器官组织中线粒体的损伤情况,探讨他克莫司(FK506)的应用对受损线粒体的作用。方法将雄性SD大鼠随机分为正常对照组、截肢后6h组、FK506干预组,每组8只。通过手术制备大鼠左后肢截肢模型。大鼠处死后迅速提取肝、肾组织线粒体,测定呼吸控制率、膜电位及三磷酸腺苷(ATP)酶活性,留取截肢后6h组肝、肾组织,电镜观察线粒体的结构。结果电镜下可见截肢后6h组大鼠肝、肾实质细胞形态、结构基本正常,但细胞内线粒体出现一定程度的损伤。与正常对照组比较,截肢后6h组肝、肾组织细胞呼吸控制率(6.09±1.18、4.77±0.65vs2.76±0.82、3.07±0.94)、膜电位(247.83±12.68、246.65±15.21vs203.23±21.51、204.54±13.40)及ATP酶活性(14.01±2.41、11.60±2.42vs8.36±1.92、6.77±1.26)显著降低(P<0.01),FK506干预组肝、肾以上指标(呼吸控制率:3.74±0.71、4.54±0.73;膜电位:243.84±7.07、245.40±6.63,ATP酶活性:10.04±1.77、10.12±2.01)高于截肢后6h组,线粒体结构损伤减轻。结论截肢创伤可导致大鼠肝、肾线粒体出现结构和功能的损伤,FK506的应用可使线粒体功能明显改善,从而推测其可能通过细胞的能量代谢途径,参与截肢创伤所致肝、肾组织损伤的修复。

Objective To investigate the hepatic and renal mitochondrial injury of rats suffering from amputation and the protective effect of tacrolimus （FK506）on chondriosome. Methods Twenty four male SD rats were divided into 3 groups at random （n=8）：control group, 6-hour-after-amputation group, and FK506 intervening group （0.1 mg/kg）. The rats were sacrificed 6 hours after trauma, and the mitochondria were extracted from livers and kidneys. The mitochondrial respiratory control ratio（RCR）,membrane potentials and total ATPase activity were detected. The morphological changes of mitochondria were observed with electron microscopy. Results Under an electron microscope, 6 hours after amputation, rats liver and kidney cells morphology and structure were normal, but there was a degree of injury within the mitochondria. Compared with control group, the mitochondrial RCR, membrane potentials and total ATPase activity decreased markedly in amputation group（RCR in liver：C 6.09±1.18,B6 2.76±0.82;RCR in kidney： C 4.77±0.65, B6 3.07±0.94 ; MP in liver： C 247.83± 12.68, B6 203.23±21.51 ; MP in kidney ： C 246.65±15.21, B6 245.40±6.63 ; ATPase in liver： C 14.01±2.41, B6 8.36±1.92 ;ATPase in kidney ： C 11.60±2.42, B6 6.77±1.26） （P〈0.01）, but they were increased in FKS06 intervening group compared with amputation group （RCR ： liver 3.74±0.71, kidney 4.54±0.73 ; MP： liver 243.84±7.07, kidney 245.40± 6.63;ATPase：liver 10.04±1.77 ,kidney 10.12±2.01 ）. Similarly, the electron microscopy showed that the morphological changes of mitochondria in FK506 intervening group were more marked than that in the amputation group. Conclusions The trauma of amputation may result in the injury of hepatic and renal mitochondria,which can be alleviated by FKS06 in a certain extent.