低温环境复灌对兔肾缺血-再灌注损伤保护作用的实验研究

丁汉东; 廖贵益; 钟金彪; 赵飞

doi:10.3969/j.issn.1674-7445.2018.05.009

低温环境复灌对兔肾缺血-再灌注损伤保护作用的实验研究

doi: 10.3969/j.issn.1674-7445.2018.05.009

230022 合肥，安徽医科大学第一附属医院泌尿外科安徽医科大学泌尿外科研究所泌尿生殖系统疾病安徽省重点实验室

基金项目:

安徽省自然科学基金 1508085SMH226

详细信息

作者简介:
丁汉东，男，1990年生，硕士，住院医师，研究方向为肾移植，Email：dinghandong1@sina.com

通讯作者:
廖贵益，男，1973年生，博士，主任医师，研究方向为肾移植，Email：liaoguiyi2@sina.com

中图分类号: R617, R692.5
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出版历程
- 收稿日期: 2018-06-10
- 网络出版日期: 2021-01-19
- 刊出日期: 2018-09-15

Experimental study of protective effect of hypothermic reperfusion on renal ischemia-reperfusion injury in rabbits

Department of Urology, First Affiliated Hospital of Anhui Medical University, Institute of Urology, Anhui Medical University, Anhui Province key Laboratory of Genitourinary Diseases, Hefei 230022, China

More Information

Corresponding author: Liao Guiyi, Email: liaoguiyi2@sina.com

摘要

摘要: 目的探讨建立兔肾缺血低温环境、常温环境及高温环境再灌注损伤模型的新方法, 并评价低温环境复灌对兔肾缺血-再灌注损伤(IRI)的影响。方法将60只健康新西兰兔随机分为5组:对照组(A组)、假手术组(B组)、低温环境复灌组(C组)、常温环境复灌组(D组)、高温环境复灌组(E组), 每组12只。术后7 d内每日检测各组兔的血清肌酐(Scr)、血尿素氮(BUN)水平; 术后1 d检测各组兔肾组织内丙二醛(MDA)含量和超氧化物歧化酶(SOD)活性; 术后1 d采用苏木素-伊红(HE)染色观察肾组织病理学变化; 术后1 d采用dUTP缺口末端标记(TUNEL)染色评价细胞凋亡。结果术后1 d, 与A组和B组比较, C、D和E组兔的Scr和BUN水平均升高(均为P < 0.01);与C组比较, D组和E组兔的Scr和BUN水平升高更明显(均为P < 0.05)。术后7 d内, C、D和E组兔的Scr和BUN水平呈下降趋势。与D组和E组比较, C组兔的Scr和BUN水平较低(均为P < 0.05)。与A组和B组比较, C、D和E组的MDA含量均升高, SOD活性均降低(均为P < 0.01);与C组比较, D组和E组的MDA含量升高更明显, SOD活性更低(均为P < 0.01)。术后1 d肾组织病理学检查示A组和B组肾组织形态结构正常, D组和E组损伤表现明显, 与D、E组比较, C组损伤较轻。TUNEL染色结果显示, D组和E组肾小管上皮细胞阳性细胞明显增多, 管腔内也可见到阳性细胞, C组阳性细胞数量较D组和E组明显减少。结论冰泥覆盖肾脏、37 ℃生理盐水及40 ℃生理盐水连续滴加肾脏可建立不同温度环境复灌模型。低温环境复灌对肾IRI具有保护作用。
- 肾脏 /
- 缺血-再灌注损伤 /
- 冷缺血 /
- 低温环境复灌 /
- 常温环境复灌 /
- 高温环境复灌 /
- 兔 /
- 凋亡
Abstract: Objective To explore a novel method for establishing rabbit models with renal ischemia-reperfusion injury (IRI) under hypothermic, normothermic and hyperthermic environments and evaluate the effect of the reperfusion under hypothermic environment on renal IRI in rabbits. Methods Sixty healthy New Zealand rabbits were randomly and evenly divided into five groups:control group (A group, n=12), sham operation group (B group, n=12), hypothermic reperfusion group (C group, n=12), normothermic reperfusion group (D group, n=12) and hyperthermic reperfusion group (E group, n=12).The serum creatinine (Scr) and blood urea nitrogen (BUN) levels were measured daily within postoperative 7 d.The content of malondialdehyde (MDA) and the activity of superoxide dismutase (SOD) in the renal tissues were measured at postoperative 1 d.Hematoxylin-eosin (HE) staining was performed to observe the histopathological changes at postoperative 1 d.Terminal deoxynucleotidyl transferase (TdT)-dUTP nick-end labeling (TUNEL staining) was adopted to detect the cellular apoptosis at postoperative 1 d. Results At 1 d after operation, the Scr and BUN levels in C, D and E groups were significantly higher than those in A and B groups (all P < 0.01).Compared with C group, the Scr and BUN levels were more notably increased in D and E groups (both P < 0.05).Within 7 d after surgery, the Scr and BUN levels tended to decline in C, D and E groups.Compared with D and E groups, the Scr and BUN levels were significantly lower in C group (both P < 0.05).Compared with A and B groups, the MDA contents were considerably increased, whereas the SOD activities were significantly decreased in C, D and E groups (all P < 0.01).Compared with C group, the MDA content was more significantly enhanced, whereas the SOD activity was remarkably decreased in D and E groups (both P < 0.01).At postoperative 1 d, the pathological examination of the renal tissues demonstrated that the renal morphology was normal in A and B groups, whereas significant renal damage was observed in D and E groups.Compared with D and E groups, the severity of renal injury was less in C group.TUNEL staining revealed that the quantity of positive renal tubular epithelial cells was significantly increased in D and E groups, and positive cells were also seen in the lumen.The quantity of positive cells in C group was significantly lower than those in D and E groups. Conclusions The reperfusion rabbit models under different temperature environments can be established by covering the kidney by ice soil and persistent administration of 37 ℃ and 40 ℃ saline.Hypothermic reperfusion exerts a protective effect on renal IRI.
- Kidney /
- Ischemia-reperfusion injury /
- Cold ischemia /
- Hypothermic reperfusion /
- Normothermic reperfusion /
- Hyperthermic reperfusion /
- Rabbit /
- Apoptosis

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图 1 术后7 d内各组兔的Scr和BUN水平的变化

Figure 1. Changes of Scr and BUN levels of rabbits in each group within 7 d after operation

下载: 全尺寸图片幻灯片

图 2 各组兔术后1 d肾组织病理学检查图片及Paller评分的比较

A~E图分别为对照组、假手术组、低温环境复灌组、常温环境复灌组、高温环境复灌组肾组织病理学图片(HE, ×200);F图为各组病理学图片Paller评分的比较, 与C组比较, ^aP < 0.01

Figure 2. Renal histopathological images and comparison of Paller scores of rabbits among each group at 1 d after operation

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图 3 各组兔术后1 d肾组织凋亡染色图片(TUNEL, ×200)

A~C图为对照组; D~F图为假手术组; G~I图为低温环境复灌组; J~L图为常温环境复灌组; M~O图为高温环境复灌组; 各组阳性细胞(绿色荧光)主要见于肾小管上皮及管腔

Figure 3. Apoptotic staining images of renal tissues of rabbits among each group at 1 d after operation

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表 1 术后1 d各组兔肾组织MDA和SOD水平的比较

Table 1. Comparison of MDA and SOD levels in renal tissues of rabbits among each group at 1 d after operation(x±s)

组别	n	MDA(nmol/mg prot)	SOD(U/mg prot)
A组	6	1.30±0.06	618±35
B组	6	1.22±0.09	623±29
C组	6	2.44±0.18^{a, b}	495±34^{a, b}
D组	6	3.45±0.34^{a, b, c}	409±32^{a, b, c}
E组	6	6.96±0.36^{a, b, c}	203±47^{a, b, c}
与A组比较, ^aP < 0.01;与B组比较, ^bP < 0.01;与C组比较, ^cP < 0.01

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