韩晓曼, 陈健文, 王雷, 王力, 张帆. 间歇性肾门阻断减轻糖尿病小鼠肾脏缺血再灌注损伤的作用研究[J]. 解放军医学院学报. DOI: 10.12435/j.issn.2095-5227.2024.125
引用本文: 韩晓曼, 陈健文, 王雷, 王力, 张帆. 间歇性肾门阻断减轻糖尿病小鼠肾脏缺血再灌注损伤的作用研究[J]. 解放军医学院学报. DOI: 10.12435/j.issn.2095-5227.2024.125
HAN Xiaoman, CHEN Jianwen, WANG Lei, WANG Li, ZHANG Fan. Effect of intermittent renal hilum occlusion on renal ischemia-reperfusion injury in diabetic mice[J]. ACADEMIC JOURNAL OF CHINESE PLA MEDICAL SCHOOL. DOI: 10.12435/j.issn.2095-5227.2024.125
Citation: HAN Xiaoman, CHEN Jianwen, WANG Lei, WANG Li, ZHANG Fan. Effect of intermittent renal hilum occlusion on renal ischemia-reperfusion injury in diabetic mice[J]. ACADEMIC JOURNAL OF CHINESE PLA MEDICAL SCHOOL. DOI: 10.12435/j.issn.2095-5227.2024.125

间歇性肾门阻断减轻糖尿病小鼠肾脏缺血再灌注损伤的作用研究

Effect of intermittent renal hilum occlusion on renal ischemia-reperfusion injury in diabetic mice

  • 摘要:
    背景  合并糖尿病的肾肿瘤患者在保留肾单位手术(nephron sparing surgery,NSS)后慢性肾病发病率高,临床缺乏有效预防及干预手段。
    目的  研究利用小鼠模型,探讨合并糖尿病在不同热缺血时间梯度下对小鼠肾脏缺血再灌注 (ischemia reperfusion,IR)损伤的影响,并分析间歇性肾门阻断减轻糖尿病小鼠肾脏缺血再灌注损伤的作用。
    方法  (1)雄性C57BL/6J糖尿病小鼠和普通小鼠各36只,分别随机分为假手术组和5个单侧肾脏IR损伤组:IR-15 min组、IR-20 min组、IR-25 min组、IR-30 min组、IR-35 min组,每组小鼠6只,再灌注24 h后通过功能学、组织学、免疫学等方法评估急性肾损伤、细胞凋亡及氧化应激损伤水平,分析不同缺血时间下糖尿病对小鼠肾脏IR损伤的影响。(2)采用雄性C57BL/6J糖尿病小鼠18只,随机分为3组:持续肾门阻断组(30 min)、间歇性肾门阻断组(在30 min缺血过程中给与5 min开放血流,周期为15-5-15 min)、假手术组,每组小鼠6只,再灌注24 h后评估急性肾损伤、细胞凋亡及氧化应激损伤水平。
    结果  (1)缺血时间为15 min时,糖尿病小鼠与普通小鼠术后均未观察到明显的肾脏功能损伤、病理损伤和氧化应激损伤;缺血时间为20 min和25 min时,糖尿病小鼠的各项肾脏功能损伤、病理损伤和氧化应激损伤指标均高于普通小鼠;缺血时间为30 min和35 min时,糖尿病小鼠与普通小鼠术后肾脏功能与病理损伤均为重度,两组间差异无统计学意义(P>0.05)。(2)相较于持续性肾门阻断,间歇性肾门阻断可降低术后24 h血肌酐(P<0.001)、急性肾小管坏死评分(P<0.001)、免疫组化NGAL半定量评分(P<0.001)、TUNEL凋亡评分(P<0.001)、丙二醛水平(P<0.01)和恢复过氧化氢酶活力(P<0.001)。
    结论  缺血时间较短时,糖尿病不加重小鼠肾脏IR后急性肾损伤,缺血时间超过20 min后,合并糖尿病加重了小鼠肾脏IR后急性损伤。间歇性肾门阻断可大幅减轻糖尿病小鼠IR后急性肾损伤,抑制氧化应激损伤发生,提示间歇性肾门阻断可能是肾肿瘤合并糖尿病患者在NSS中减轻外科缺血性肾损伤的有效手段。

     

    Abstract:
    Background  Patients with renal tumor complicated with diabetes mellitus have a high incidence of chronic kidney disease after nephron sparing surgery (NSS), and lack of effective prevention and intervention in clinic.
    Objective  To investigate the effect of diabetes mellitus on renal ischemia-reperfusion (IR) injury in mice under different warm ischemia time gradients and analyze the effect of intermittent renal hilum occlusion on reducing renal ischemia-reperfusion injury in diabetic mice.
    Methods  Thirty-six nondiabetic male mice and thirty-six diabetic C57BL/6J male mice were randomly divided into six groups, ischemia reperfusion (IR)-15 minutes, IR-20, IR-25, IR-30, IR-35 minutes and sham group (n = 6 for each group). Unilateral kidney ischemia-reperfusion injury mouse model was established, after 24 h reperfusion, the levels of acute kidney injury, apoptosis and oxidative stress injury were evaluated by functional, histological and immunological methods, and the effects of diabetes on renal IR injury in mice under different ischemic time points were analyzed. Eighteen C57BL/6J male mice were randomly divided into continuous hilar occlusion group, intermittent hilar occlusion group (15 minutes of ischemia followed by 5 minutes of reperfusion, 15-5-15 minutes), and sham group (n = 6 for each group). The levels of acute kidney injury, apoptosis and oxidative stress injury were assessed via functional and histological analysis after 24 h reperfusion.
    Results  When the ischemia time was 15min, no obvious renal function injury, pathological injury and oxidative stress injury were observed in diabetic mice and normal mice after operation; When the ischemia time was 20min and 25min, the indexes of renal function injury, pathological injury and oxidative stress injury in diabetic mice were higher than those in normal mice; When the ischemia time was 30min and 35min, the postoperative renal function and pathological injury of diabetic mice and ordinary mice were severe, and there was no significant difference between the two groups (P>0.05). Compared with continuous renal hilum occlusion, intermittent renal hilum occlusion could reduce serum creatinine (P < 0.001), acute tubular necrosis score (P < 0.001), immunohistochemical NGAL semi quantitative score (P < 0.001), TUNEL apoptosis score (P < 0.001), malondialdehyde level (P < 0.01) and restore catalase activity (P < 0.001) at 24 h after operation.
    Conclusion  Diabetes mellitus does not aggravate acute kidney injury after IR in mice with short ischemic time. After ischemia time exceeding 20min, diabetes mellitus aggravates acute kidney injury after IR in mice. Intermittent renal hilum occlusion can significantly reduce the acute kidney injury after IR in diabetic mice and inhibit the occurrence of oxidative stress injury, suggesting that intermittent renal hilum occlusion may be an effective means of reducing surgical ischemic kidney injury in patients with renal tumor and diabetes in NSS.

     

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