摘要
目的建立山羊挤压综合征(CS)动物模型,并探讨连续性静脉-静脉血液滤过(CVVH)治疗对肾功能及肾组织病理变化的影响。方法 12只山羊随机分为对照组、CS模型组、CVVH治疗组,每组4只。CS模型组与CVVH治疗组山羊于后肢肌肉注射50%甘油生理盐水溶液10 mL/kg建立CS模型(对照组注射等量生理盐水),1、2、8、12、24h后检测血清肌酸激酶(sCK)和血肌酐(sCr),以sCK>1 000U/L,同时sCr>2倍对照值判断为造模成功。造模1h后,CVVH治疗组于造模对侧股静脉留置单针双腔导管建立血管通路,采用智能化床旁肾脏替代治疗机进行CVVH治疗。血流量为100mL/min,置换液流速为35mL/(kg·h),以前稀释法输入,治疗23h后(第24h)处死动物,留取肾组织标本,光镜、电镜下观察肾组织病理组织结构变化;采用免疫组化染色检查caspase12蛋白表达变化及TUNEL染色对细胞凋亡进行检测。结果造模山羊均于注射甘油后1h内出现酱油色小便,尿量较对照组明显减少。CS模型组与CVVH治疗组造模1h后血清sCr及sCK较对照组升高,差异有统计学意义(P<0.05),提示造模成功。行CVVH治疗23h(第24h),sCK、sCr水平低于CS模型组(P<0.05)。光镜下CS模型组肾组织可见急性肾小管坏死表现,肾间质水肿,肾小球基本正常。电镜下模型组可见小管上皮细胞明显的染色质聚集,线粒体肿胀,内质网扩张等早期细胞凋亡征象,CVVH治疗组表现稍轻。CS模型组及CVVH治疗组caspase12表达高于对照组(P<0.001),CVVH治疗组caspase12表达低于CS模型组(P<0.05)。TUNEL染色证实CS模型组山羊肾组织细胞凋亡比例明显增高,CVVH治疗组较模型组凋亡比例少。结论通过肌肉注射50%甘油生理盐水溶液可建立山羊CS模型,早期行CVVH治疗可减缓肾功能恶化,减轻肾小管上皮细胞凋亡程度。
Objective Crush syndrome（CS）is a common critical condition.This study aimed to establish a crush syndrome goat model through muscle injection of glycerol and test the effect of continuous veno-venous hemofiltraion（CVVH）.Methods 12 male goats at 12-15 months age were randomly assigned into control,model,and CVVH groups.After 2weeks of normal feeding,the goats were weighed and stripped off foods for 24 h.Goats in the model and CVVH groups were then injected with 50% glycerol athind legs.Diagnosis of CS was established based on serum creatine kinase（CK）1 000 U/L and serum creatinine（sCr） 2times of standard.No intervention was given to goats in the control group.A catheter was planted to get blood access.CVVH was administered using a Prisma-flex machine,with blood flow being set at 100mL/min and replacement fluid with predilution at 35mL/（kg·h）.After 23 hof treatment,the goats were sacrificed.Plasma and kidney samples were taken.Results Bloody urine hyperkalemia and decrease of urine volume were found in all of the goats injected with glycerol.Serum CK and sCr increased 1hafter the injection compared with the controls.After 23 hof CVVH treatment,serum CK and sCr decreased compared with goats in the model group.The light microscope revealed manifestation of tubular necrosis and interstitial edema,but the glomeruli were almost normal.The electronic microscope found prominent signs of cell apoptosis,such as chromatin aggregation,mitochondrial swelling,and endoplasmic reticulum expansion.Caspase12 expression in the goats with CS was significantly higher than that in the controls.The CVVH treated goats had lower level of expression than those in the model group（P〈0.05）.TUNEL staining identified a higher proportion of renal cell apoptosis in the goats in the model group compared with those in the CVVH group.Conclusion Muscle injection with glycerol can induce CS in goats.Early CVVH intervention improves renal function and alleviates renal tubular cell apoptosis.
出处
《四川大学学报（医学版）》
CAS
CSCD
北大核心
2016年第1期28-32,共5页
Journal of Sichuan University(Medical Sciences)
基金
国家自然科学基金(No.81270818)资助
