目的 探讨大黄萃取液对心搏骤停(CA)兔心肺复苏(CPR)后肾损伤的预防作用及机制.方法 将24只雄性日本大耳白兔按随机数字表法分为假手术组、复苏模型组和大黄预处理组,每组8只.大黄预处理组在制模前连续灌胃大黄萃取液5 mL·kg-1·d-17 d;假手术组、复苏模型组则连续灌胃生理盐水10 mL/d 7 d.连续干预7 d后针刺心底和心尖部,用50 V交流电刺激诱发心室纤颤(室颤)制备CA-CPR动物模型.于复苏成功后2 h处死复苏模型组和大黄预处理组动物,假手术组动物于麻醉后直接处死;用全自动生化分析仪检测血清尿素氮(BUN)和肌酐(Cr)水平,用免疫组化法检测肾组织中性粒细胞明胶酶相关脂质运载蛋白(NGAL)和白细胞介素-18(IL-18)的蛋白表达.结果 与假手术组比较,复苏模型组和大黄预处理组血清BUN、Cr水平均明显增加〔BUN(mmol/L):15.53±3.90、10.51±3.16比7.03±2.23,Cr(μmol/L):137.20±12.23、86.80±7.67比66.39±5.47,均P〈0.05〕;而大黄预处理组血清BUN和Cr水平则较复苏模型组明显降低〔BUN(mmol/L):10.51±3.16比15.53±3.90,Cr(μmol/L):86.80±7.67比137.20±12.23,均P〈0.05〕.免疫组化染色显示,肾组织NGAL、IL-18主要表达于肾小球及肾小管细胞胞质中,复苏模型组和大黄预处理组阳性表达面积均较假手术组明显增加〔NGAL(μm2):208.26±7.58、136.74±5.33比98.93±7.83,IL-18(μm2):256.48±4.64、113.22±6.98比77.06±6.47,均P〈0.05〕;而大黄预处理组NGAL、IL-18表达均较复苏模型组明显减少 〔NGAL(μm2):136.74±5.33比208.26±7.58,IL-18(μm2):113.22±6.98比256.48±4.64,均P〈0.05〕.结论 CA可导致急性肾损伤(AKI)的发生;大黄萃取液可减少CPR后兔肾组织中NGAL、IL-18表达,对CPR后肾脏具有保护作用.
Objective To study the preventive effect and mechanism of Rhubarb extract on renal injury after cardiopulmonary resuscitation (CPR) in rabbits with cardiac arrest (CA).Methods Twenty-four male Japanese big-ear rabbits were divided into sham operation group, CPR model group and Rhubarb pretreatment group by random number table method, with 8 rabbits in each group. The rabbits in Rhubarb pretreatment group were treated with Rhubarb extract 5 mL·kg-1·d-1 for 7 days; and those in sham operation group and CPR model group were fed with 0.9% normal saline 10 mL/d for 7 days. After 7 days, ventricular fibrillation (VF) was produced in CPR model group and Rhubarb pretreatment group by 50 V alternating currents stimulation through bottom of the heart leads to the apex to prepare CPR model. The rabbits of the CPR model group and Rhubarb pretreatment group were sacrificed at 2 hours after successful resuscitation, and the animals in the sham operated group were sacrificed directly after anesthesia. The levers of blood urea nitrogen (BUN) and creatinine (Cr) in serum were examined by automatic biochemical analyzer. The positive expression area of neutrophil gelatinase-associated lipocalin (NGAL) and interleukin-18 (IL-18) in kidney were examined by immunohistochemistry.Results Compared with the sham operation group, the levels of BUN and Cr were significantly increased in the CPR model group and Rhubarb pretreatment group [BUN (mmol/L):15.53±3.90, 10.51±3.16 vs. 7.03±2.23, Cr (μmol/L): 137.20±12.23, 86.80±7.67 vs. 66.39±5.47, bothP 〈 0.05]. Compared with the CPR model group, the levels of BUN and Cr were significantly decreased in the Rhubarb pretreatment group [BUN (mmol/L): 10.51±3.16 vs. 15.53±3.90, Cr (μmol/L): 86.80±7.67 vs. 137.20±12.23, bothP 〈 0.05]. Immunohistochemical staining showed that the expression of NGAL and IL-18 mainly existed in glomerular and tubular cells in patina. Compared with the sham operation group, the positive expre