目的 建立蜂毒肽所致急性肾损伤（AKI）的山羊模型，并观察连续性静-静脉血液滤过（CVVH）的干预效果及可能机制。 方法 将雄性杂交山羊随机分为3组：空白对照组，蜂毒肽致AKI模型组及CVVH干预组，每组4只。蜂毒肽按照总剂量0.5 mg/kg在48 h内分为4次由耳缘静脉注入后两组动物体内，空白对照组注入等量生理盐水。第1次注射后每6 h采血检测血清肌酐（sCr）、肌酸激酶（CK）等，记录每小时尿量。检测到sCr大于2倍对照值或尿量少于0.5 mL/（kg·h）超过6 h判断造模成功。造模成功后CVVH组于右侧股静脉建立血管通路，采用普通肝素抗凝行CVVH治疗12 h。随后麻醉处死各组动物，以开放式肾活检获取肾脏标本，进行光镜及电镜观察，采用免疫组化及TUNEL染色检测凋亡。 结果 第1次注射后48 h时sCr检测值〔模型组（100.75±7.87） μmol/L vs. 对照组（43.95±1.59） μmol/L vs. CVVH组(102.10±5.06) μmol/L，P<0.001〕示造模成功。CVVH干预后sCr较模型组降低 〔(64.13±5.82) μmol/L vs. (108.60±9.40) μmol/L，P<0.001〕。光镜观察示CVVH组肾脏组织病理学改变明显轻于模型组。电镜观察示与对照组相比，模型组肾小管上皮细胞胞质出现空泡样变，线粒体等细胞器肿胀较明显，CVVH亦有类似改变，但程度较模型组轻。免疫组化染色示模型组肾组织凋亡标志蛋白caspase-3表达高于CVVH组及对照组。TUNEL染色提示模型组肾组织凋亡细胞明显高于对照及干预组（2.48%±0.11% vs. 0.08%±0.01% vs. 1.40%±0.12%，P<0.001）。 结论 成功建立了蜂毒肽致AKI的山羊模型并顺利完成CVVH干预。CVVH干预可减轻蜂毒肽所致AKI，其机制或与肾小管上皮细胞凋亡有关。
英文摘要:
Objective To establish a goat model of melittin induced acute kidney injury (AKI), and to evaluate the therapeutic effect of continuous veno-venus heamofiltration (CVVH) in melittin- induced AKI. Methods Twelve male goats were randomized into three groups: control group, melittin induced AKI group (melittin group), and CVVH intervention group (CVVH group). The AKI goat model was established by the injection of melittin via the auricular vein for four times in 48 h to reach a total dose of 0.5 mg/kg, then serum creatinine (Cr) and creatine kinase (CK) were tested every 6 h and urine output was record each hour. AKI was diagnosed when Cr level increased to the double value of control group, or the urine output decreased to less than 0.5 mL/(kg·h) in 6 h. After the diagnosis of AKI, the animals in CVVH group received CVVH treatment for 12 h. At the end, the goats in all groups were sacrificed by anesthesia and kidney tissue samples were collected. Light microscopy and telectron microscopy observation were performed. Apoptosis was detected by immunohistochemistry and TUNEL technique. Results AKI was successfully induced by melittin in the goats. The Cr level in control group was (43.95±1.59) μmol/L, while (100.75±7.87) μmol/L in AKI group and (102.10±5.06) μmol/L in CVVH group. Cr level was lowered significantly after CVVH treatment 〔(45.02±2.41) μmol/L in control group vs. (108.60±9.40) μmol/L in AKI group vs. (64.13±5.82) μmol/L in CVVH group, P<0.001〕. Swelling and reduction of mitochondrial crests in AKI group were more obvious than those in CVVH group. Expression of caspase-3 and apoptosis cells percentage of renal tubules in AKI group were significantly higher than those in CVVH group. Conclusion Melittin induced AKI model could be established in goats. CVVH could alleviate melittin induced AKI, probably in the mechanism to reduce the apoptosis of renal tubular cells.
