不同液体复苏方式对失血性休克大鼠各器官细胞凋亡的影响

目的观察大鼠失血性休克时各器官细胞的凋亡情况及不同液体复苏方式对其影响。方法将32只SD大鼠制作成未控制性重度失血性休克模型,随机分为对照组、无液体复苏组(NF组)、限制性液体复苏组(NS40组)和快速大量液体复苏组(NS80组),每组8只,比较各组的救治疗效,并采用光镜、流式细胞仪和末端脱氧核苷酸转移酶介导的原位末端标记法(TUNEL)检测失血/复苏/急救后各组存活大鼠的肝、肾、肺和小肠黏膜等脏器细胞凋亡。结果NS40、NS80两组液体复苏(14/16)大鼠的存活率较NF组(3/8)明显增高(P<0.05),而血清乳酸水平明显降低(P<0.01)。各组存活大鼠的肝、肾、肺和小肠黏膜等器官经TUNEL法染色,在荧光显微镜下均可见发绿色荧光阳性细胞,其中NS80组存活大鼠的肾(2.6±0.8个/高倍视野)和小肠黏膜(48.0±4.0个/高倍视野)等器官中凋亡阳性细胞数明显多于NF组(分别为1.3±0.7个/高倍视野、32.4±4.4个/高倍视野)和NS40组(分别为1.3±0.6个/高倍视野、37.8±5.6个/高倍视野),差异具有统计学意义。同时,流式细胞仪检测显示,NS80组大鼠的肝(0.74%±0.24%)、肾(4.62%±1.19%)和小肠黏膜(9.98%±1.01%)等器官的细胞凋亡率明显高于NF组(分别为0.44%±0.14%、1.29%±0.18%、2.55%±0.28%)和NS40组(分别是0.34%±0.12%、1.67%±1.1

Effects of different means of fluid resuscitation on apoptosis of visceral organs in rats with hemorrhagic shock

Objective To observe the effects of different means of fluid resuscitation on apoptosis of visceral organs in rats with hemorrhagic shock.Methods The tails of 8 male SD rats were cut to cause active bleeding. Blood was collected from the carotid arteries of another 24 male SD rats and heparinized, then the 24 rats were randomly divided into 3 equal groups: no fluid resuscitation group (NF group, carotid blood was collected as described above, the tail was cut and the blood from the tail was collected in container with heparin 30 min after, and hemostasis and heparin blood transfusion were performed 60 min after cutting of the tail), controlled fluid resuscitation group [NS40 group: isosmotic saline was infused during the period of 30 to 60 min after the tail cutting to maintain the mean arterial pressure (MAP) at about 40 mm Hg and then hemostasis and heparin blood transfusion were performed 60 min after cutting of the tail], and great quantity fluid rapid resuscitation group [NS80 group: a great quantity of isosmotic saline was infused during the period of 30 to 60 min after the tail cutting to maintain the MAP at about 80 mm Hg and then hemostasis and heparin blood transfusion were performed 60 min after cutting of the tail]. Blood specimens were collected at the time points 0, 120, and 150 min to undergo blood routine examination. Another blood specimens were collected at the time points 0, 30, 60, and 90 min to undergo lactic acid examination. The surviving rats were killed and their livers, kidneys, lungs, and small intestines were taken out to undergo pathology. Cell apoptosis was examined by flow cytometry and TUNEL.Results The survival rates of the NS40 and NS80 groups were significantly higher than that of the NF group (both P<0.05). The blood lactic acid levels of the NS40 and NS80 groups at the time points 60 and 90 min were all significantly lower than those of the NF and control groups (all P<0.05). Apoptosis in the liver, kidney, and small intestine mucosa of the NS80 group was significantly marked than in the NF and NS40 groups (all P<0.01).Conclusion Controlled fluid resuscitation obviously reduces the early death rate of rats with severe hemorrhagic shock and apoptosis in the liver, kidney, and small intestine mucosa thereof and may benefit the prognosis.