摘要/Abstract
摘要： 目的·观察姜黄素对心肺复苏（cardiopulmonary resuscitation，CPR）后大鼠肠缺血/再灌注（ischemia/reperfusion，I/R）损伤的影响，并初步探讨其作用机制。方法·24只健康雄性SD大鼠随机分为4组，每组6只，即对照组（Sham组）、姜黄素组（Cur组）、CPR后肠I/R损伤组（I/R组）和姜黄素+CPR后肠I/R损伤组（Cur+I/R组）。4个组均行麻醉后气管插管，股动、静脉置管；I/R组和Cur+I/R组通过窒息法建立心脏停搏/CPR模型，Sham组和Cur组不进行该操作；Cur+I/R组于CPR开始前1 min腹腔注射姜黄素100 mg/kg，同一时间Cur组腹腔注射等量姜黄素，Sham组、I/R组注射同等体积二甲基亚砜。自主循环恢复后6、12、24 h静脉采血检测肿瘤坏死因子α（tumor necrosis factor-α，TNF-α）、白介素6（interleukin-6，IL-6）的浓度；自主循环恢复后24 h处死大鼠，检测肠组织中丙二醛（malondialdehyde，MDA）、还原型谷胱甘肽（glutathione，GSH）浓度和超氧化物歧化酶（superoxide dismutase，SOD）活力，苏木精-伊红染色观察肠组织病理学变化，TUNEL法检测肠黏膜上皮细胞凋亡情况，Western blotting法检测肠组织中缺氧诱导因子1α（hypoxia-inducible factor-1α，HIF-1α）、胱天蛋白酶3（caspase-3）的表达情况，透射电子显微镜下观察肠组织超微结构改变。结果·Cur+I/R组3个时间点的TNF-α、IL-6的浓度均低于I/R组（均P<0.05），高于Sham组（均P<0.05）。Cur+I/R组的MDA含量低于I/R组（P<0.01），高于Sham组（P<0.01）；Cur+I/R组的SOD活力和GSH浓度高于I/R组（均P<0.05），低于Sham组（均P<0.01）。Cur+I/R组的HIF-1α蛋白表达水平显著高于I/R组（P<0.01），低于Sham组（P<0.01）；而Cur+I/R组caspase-3蛋白表达水平显著低于I/R组（P<0.01），高于Sham组（P<0.01）。病理学观察发现，I/R组可见回肠黏膜水肿，炎症细胞浸润，绒毛顶端上皮脱落、粘连，固有层裸露、破溃，而Cur+I/R组肠黏膜损伤较I/R组明显减轻。TUNEL法检测显示，Cur+I/R组凋亡指数较I/R组显著降低（P<0.01），但较Sham组升高（P<0.01）。电子显微镜下Sham组、Cur组肠黏膜形态正常，I/R组肠黏膜上皮细胞微绒毛排列稀疏、倒伏，内质网脱颗粒，线粒体空化、嵴断裂；Cur+I/R组肠黏膜上皮细胞微绒毛排列密集、有序，结构清晰，细胞间的连接复合体完整，细胞器形态大致正常。结论·姜黄素对CPR后大鼠肠黏膜I/R损伤具有抑制作用，其机制与抑制炎症、降低氧化应激、抗凋亡及上调HIF-1α蛋白表达有关。
关键词: 心肺复苏, 姜黄素, 肠黏膜, 缺血/再灌注损伤, 氧化应激, 炎症, 凋亡
Abstract:
Objective · To observe the effect of curcumin on intestinal ischemia/reperfusion (I/R) injury in the rats after cardiopulmonary resuscitation (CPR), and explore the mechanism. Methods · Twenty-four healthy male SD rats were randomly divided into 4 groups, namely control group (Sham group, n=6), curcumin group (Cur group, n=6), intestinal I/R injury after CPR group (I/R group, n=6), and curcumin+intestinal I/R injury after CPR group (Cur+I/R group, n=6). Tracheal and femoral artery and vein catheterization were carried out after anesthesia in the 4 groups. The cardiac arrest/CPR model was established by asphyxiation in I/R group and Cur+I/R group, while not in the other two groups. Curcumin (100 mg/kg) was injected intraperitoneally 1 min before CPR in Cur+I/R group. Meanwhile, equivalent curcumin was injected intraperitoneally in Cur group, and equivalent dimethylsulfoxide was injected in Sham group and I/R group. Venous blood samples were collected 6, 12 and 24 h after recovery of spontaneous circulation (ROSC) to detect the concentrations of tumor necrosis factor-α (TNF-α) and interleukin-6 (IL-6). The rats were sacrificed 24 h after ROSC, and the concentrations of malondialdehyde (MDA) and glutathione (GSH) and the activity of superoxide dismutase (SOD) in the intestinal tissues were measured. The pathological changes of intestinal tissues were observed by hematoxylin-eosin staining. The apoptosis of intestinal mucosal epithelial cells was detected by TUNEL assay. Hypoxia-inducible factor-1α (HIF-1α) and caspase-3 expressions in the intestinal tissues were detected by Western blotting. Ultrastructural changes of the intestinal tissues were observed by using transmission electron microscopy. Results · The concentrations of TNF-α and IL-6 in Cur+I/R group at 3 time points were lower than those in I/R group (all P<0.05), while higher than those in Sham group (all P<0.05). The MDA content in Cur+I/R group was lower than that in I/R group (P<0.01), while higher than that in Sham group (P<0.01). The SOD activity and GSH concentration in Cur+I/R group were higher than those in I/R group (both P<0.05), while lower than those in Sham group (both P<0.01). The expression of HIF-1α protein in Cur+I/R group was significantly higher than that in I/R group (P<0.01), but lower than that in Sham group (P<0.01). The expression of caspase-3 protein in Cur+I/R group was significantly lower than that in I/R group (P<0.01), while higher than that in Sham group (P<0.01). Pathological observation showed that ileal
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