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目的:通过观察小鼠止血带休克(TS)后,不同时点血管紧张素转换酶(ACE)和血管紧张素转换酶2(ACE2)在肾脏的表达变化与肾损伤程度的关系,探讨ACE/ACE2表达失衡在TS后肾损伤中的作用。方法:复制小鼠TS模型,Western blotting测肢体缺血再灌注后12 h内肾组织ACE和ACE2蛋白的表达;利用化学比色方法测定血清和肾组织丙二醛(MDA)含量和超氧化物歧化酶(SOD)活性;制作肾病理切片,观察肾组织形态变化,并利用免疫组织化学方法观察ACE和ACE2的表达部位。结果:Western blotting结果显示,各时点与对照组比较,TS后ACE表达升高,ACE2表达降低;与对照组比较血清和肾MDA水平增高(P<0.05),SOD活性降低(P<0.05);HE病理切片显示,TS后各时点肾组织有充血、炎细胞浸润等不同程度损伤;免疫组化结果显示,ACE在肾小管上皮细胞胞浆表达,TS后表达明显增强;ACE2主要在肾小管上皮细胞管腔膜表达,TS后表达明显降低。结论:TS后肾组织ACE表达升高,ACE2表达降低,ACE/ACE2表达失衡可能与肾损伤有关。
OBJECTIVE: To investigate the relationship between the expression of angiotensin converting enzyme (ACE) and angiotensin converting enzyme 2 (ACE2) in kidney and the degree of renal injury at different time points after the mice were treated with tourniquet hemorrhage (TS) The Role of Unbalanced ACE2 Expression in Renal Injury after TS. Methods: TS model was duplicated and the expressions of ACE and ACE2 protein in renal tissues within 12 h after limb ischemia-reperfusion were detected by Western blotting. The contents of malondialdehyde (MDA) and superoxide Dismutase (SOD) activity was measured. Nephridial sections were made and morphological changes of renal tissues were observed. Immunohistochemistry was used to observe the expression of ACE and ACE2. Results: Compared with the control group, the expression of ACE was increased and the expression of ACE2 was decreased after TS in each time point. Compared with the control group, the serum and kidney MDA levels increased (P <0.05) and the SOD activity decreased (P <0.05) ; HE pathological sections showed that there were hyperemia and infiltration of inflammatory cells in different degrees at different time points after TS. Immunohistochemistry showed that ACE was expressed in the cytoplasm of renal tubular epithelial cells and significantly increased after TS; ACE2 mainly localized in the kidney Tubular epithelial cell lumen membrane expression, TS expression was significantly reduced. Conclusion: After TS, the expression of ACE is increased and the expression of ACE2 is decreased. The unbalanced expression of ACE / ACE2 may be related to renal injury.