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缺血后处理通过激活PI3K/AKT通路减轻大鼠缺血/再灌注肺损伤

颜王鑫¹, 程缘², 黄曼², 徐俊鹏², 骆珍珍², 王淑远³, 戴雍月², 尤利益⁴, 王万铁^2,*

¹温州市人民医院结直肠肛门外科，温州 325400；²温州医科大学缺血/再灌注损伤研究所，温州 325035；³温州市人民医院呼吸内科，温州 325400；⁴温州市人民医院超声科，温州 325400

摘要

本文旨在探讨缺血后处理(ischemic post-conditioning, I-post-C)是否通过激活PI3K/AKT通路减轻大鼠肺缺血/再灌注损伤(lung ischemia-reperfusion injury, LIRI)。将6~8 周龄Sprague Dawley (SD)大鼠随机分为4 组：对照(Control)组、缺血/再灌注(I/R)组、I-post-C 组和I-post-C+PI3K抑制剂(I-post-C+LY294002)组。I/R 组大鼠左侧肺门用微型动脉夹夹闭30 min 后松开，再灌注1 h；I-post-C 组左侧肺门阻断30 min，再灌注30 s，如此循环3 次后再灌注1 h；I-post-C+LY294002 组大鼠术前3 天每日腹腔注射1 次LY294002 (10 mg/kg)，后续处理同I-post-C 组。用HE染色并计算总肺含水量(total lung water content, TLW)、肺组织定量评价指标(index of quantitative assessment, IQA)、肺湿干重比(lung wet-to-dry weight ratio, W/D)来评估肺组织损伤程度，用TUNEL染色检测肺组织凋亡水平，用Western blot 检测肺组织Caspase 3 活化水平及磷脂酰肌醇3-激酶(phosphatidylinositol3-kinase, PI3K)/蛋白激酶B (protein kinase B, PKB/AKT)、糖原合成酶激酶3β (glycogen synthase kinase 3β, GSK-3β)磷酸化水平，用JC-1染色检测肺组织线粒体膜电位。结果显示，与Control 组相比，I/R 组肺组织损伤明显，线粒体膜电位显著下降，cleaved Caspase 3/pro-Caspase 3 比值和凋亡水平显著升高，PI3K/AKT/GSK-3β 磷酸化水平显著下降。I-post-C 可显著逆转I/R 组肺组织的上述变化，发挥肺组织保护作用；而PI3K抑制剂LY294002 可部分消除I-post-C 的这种肺保护作用。以上结果提示，I-post-C 可能通过激活PI3K/AKT通路失活GSK-3β，阻抑线粒体膜电位下降，拮抗肺组织细胞凋亡，从而减轻LIRI。

关键词： 大鼠; 缺血后处理; 缺血/再灌注损伤; 细胞凋亡; PI3K/AKT通路

Ischemic post-conditioning alleviates I/R lung injury by activating PI3K/AKT pathway in rats

YAN Wang-Xin¹, CHENG Yuan², HUANG Man², XU Jun-Peng², LUO Zhen-Zhen², WANG Shu-Yuan³, DAI Yong-Yue², YOU Li-Yi⁴, WANG Wan-Tie^2,*

¹Department of Colorectal and Anal Surgery, Wenzhou People's Hospital, Wenzhou 325400, China；²Institute of Ischemia/Reperfusion Injury, Wenzhou Medical University, Wenzhou 325035, China；³Department of Respiratory Medicine, Wenzhou People's Hospital, Wenzhou 325400, China；⁴Department of Ultrasound, Wenzhou People's Hospital, Wenzhou 325400, China

Abstract

The present study aimed to investigate whether ischemic post-conditioning (I-post-C) could reduce lung ischemia/reperfusion injury (LIRI) by activating PI3K/AKT pathway in rats. Male Sprague-Dawley (SD) rats aged 6-8 weeks were randomly divided into four groups: control group, ischemia/reperfusion (I/R) group, I-post-C group, and I-post-C+PI3K inhibitor (I-post-C+LY294002) group. The left pulmonary hilum of rats in the I/R group was clamped for 30 min with a miniature arterial clip and then released, followed by reperfusion for 1 h; the left pulmonary hilum of rats in the I-post-C group was blocked for 30 min, followed by reperfusion for 30 s, and this process was repeated three times before reperfusion for 1 h; rats in the I-post-C+LY294002 group were intraperitoneally injected with LY294002 (10 mg/kg) once a day for 3 d before surgery, and the subsequent treatment was the same as that of the I-post-C group. Lung tissue injury was assessed using HE staining and calculation of total lung water content (TLW), index of quantitative assessment (IQA), and lung wet-to-dry weight ratio (W/D). TUNEL assay was used to detect the level of apoptosis in lung tissue. Western blot was used to detect the activation level of Caspase 3 (cleaved Caspase 3/pro-Caspase 3 ratio) and the phosphorylation levels of phosphatidylinositol 3-kinase (PI3K), protein kinase B (PKB/AKT) and glycogen synthase kinase 3β (GSK-3β) in lung tissue. Mitochondrial membrane potential was measured by JC-1 staining. The results showed that compared with the control group, the I/R group exhibited significant lung tissue damage, a significant decrease in mitochondrial membrane potential, significantly elevated cleaved Caspase 3/pro-Caspase 3 ratio and cell apoptosis level, as well as decreased phosphorylation levels of PI3K/AKT/GSK-3β. I-post-C significantly reversed the aforementioned changes in lung tissue of the I/R group and exerted a protective effect on lung tissue. The PI3K inhibitor LY294002 partially eliminated this lung protective effect of I-post-C. These results suggest that I-post-C activates the PI3K/AKT pathway to inactivate GSK-3β, inhibits mitochondrial membrane potential decline, antagonizes lung tissue cell apoptosis, and thereby alleviates LIRI.

Key words: Rats; ischemic post-conditioning; ischemia/reperfusion injury; apoptosis; PI3K/AKT pathway

收稿日期：　　录用日期：

通讯作者：王万铁　　E-mail:

DOI: 10.13294/j.aps.2026.0050

引用本文：

颜王鑫, 程缘, 黄曼, 徐俊鹏, 骆珍珍, 王淑远, 戴雍月, 尤利益, 王万铁. 缺血后处理通过激活PI3K/AKT通路减轻大鼠缺血/再灌注肺损伤[J]. 生理学报 2026; 78 (3): 705-712.

YAN Wang-Xin, CHENG Yuan, HUANG Man, XU Jun-Peng, LUO Zhen-Zhen, WANG Shu-Yuan, DAI Yong-Yue, YOU Li-Yi, WANG Wan-Tie. Ischemic post-conditioning alleviates I/R lung injury by activating PI3K/AKT pathway in rats. Acta Physiol Sin 2026; 78 (3): 705-712 (in Chinese with English abstract).