高容量血液滤过对脓毒症犬肾组织线粒体功能的影响及其可能机制

Effect and Mechanism of High Volume Hemofiltration on Mitochondria Function of Sepsis Dog Nephridial Tissue

探讨高容量血液滤过对脓毒症犬肾组织线粒体功能的影响。采用盲肠结扎穿孔术(CLP)制备脓毒症模型:将健康雄性犬30只随机分为假手术组(A组);脓毒症模型组(B组)和脓毒症+高容量血液滤过治疗组(C组)。各组分别于00、.5、1、2、3、4 h等不同时点测定心率、血压、血肌酐(Scr)、肿瘤坏死因子-α(TNF-α)浓度。于4 h点处死动物,观察肾组织病理改变及肾小管损伤程度;采用生物发光测量法检测肾组织线粒体丙二醛(MDA)、超氧化物歧化酶(SOD)活性、活性氧(ROS)、三磷酸腺苷酶(ATP酶)、ATP生成量(P/O比值)变化。结果B组动物与A组相比较,出现心率增快、血压降低;血Scr、TNF-α水平于造模后0.5 h升高,肾组织出现明显的病理损害、肾小管坏死评分值明显增高,肾组织中三磷酸腺苷(ATP)含量明显下降,氧自由基(ROS)含量升高;其中SOD酶活性的下降与TNF-α含量的增高呈现良好的相关性(r=0.65,P<0.01)。经HVHF治疗后,C组犬肾组织损伤轻于B组,相应指标亦好于B组。氧化应激反应能力的相对不足和氧化损伤在脓毒症急性肾损伤发病过程中起了重要作用;HVHF通过清除异常增高的炎症物质水平,减轻肾组织的氧化损伤,改善肾小管上皮细胞线粒体的能量代谢状态。

To approach the effect and possible mechanism of high vo lume hemofiltration（HVHF） on mitochondria function of sepsis dog nephridial tis sue.Sepsis model was established with cecum ligature perforation（CL P）,30 healthy male dogs were equally randomized into following groups： the sham-operated group（group A）,sepsis group（group B） and sepsis with HVHF treatment group（group C）.Heart rate,blood pressure,serum creatinine（Scr） and tumor n ecrosis factor-α（TNF-α） level in each group were mensured at six observation p er iods（0,0.5,1,2,3 h and 4 h）.Levels of malondialdehyde（MDA）,activity o f superoxide dismutase（SOD）,reactive oxygen species（ROS）,adenosine triphosph atase（ATP enzyme）,production of ATP（oxidative phosphorylation ratio） in mitoc hondria of nephridial tissue were tested by bioluminescent measuring method at 4 h.Compared with group A,the increased heart rate,decreased blood pressure,le vel of Scr and TNF-α step up after modeled 0.5 h in group B,also apparent p athological lesion appeared,renal tubular necrosis score obviously increased,ATP significantly decreased and ROS increased in nephridial tissue.Among them,the d escent of SOD activity and increase of TNF-α level present a good correlation （r=0.65,P0.01）.After HVHF treatment,injury of dog nephridial tissue in group C were slighter,also the index of counterpart were better than t hat group B.Relatively insufficient of oxidative stress competence an d oxidative damage play the vital role in sepsis pathogenesis of acute kidney in jury.By getting rid of the abnormal increased inflammation materials,HVHF can relieve oxidative damage in nephridial tissue and improve the energy metabolism state of mitochondria in renal tubular epithelial cell.