食欲刺激素对乙酸致大鼠胃损伤的保护作用及机制  

Protective effect and mechanism of ghrelin against gastric ulcer induced by acetic acid

作　　者：袁月 姜剑伟 唐波 王贵彬 张建军 YUAN Yue;JIANG Jian-wei;TANG Bo;WANG Gui-bin;ZHANG Jian-jun(Beijing Key Laboratory of New Drug Action Mechanism Research and Pharmacodynamic Evaluation,Institute of Materia Medica,Chinese Academy of Medical Sciences&Peking Union Medical College,Beijing 100050,China)

机构地区：[1]中国医学科学院北京协和医学院药物研究所,新药作用机制研究与药效评价北京市重点实验室,北京100050

出　　处：《中国药理学与毒理学杂志》2022年第3期177-184,共8页Chinese Journal of Pharmacology and Toxicology

基　　金：国家自然科学基金(81773715);国家自然科学基金(81630094);国家科技重大专项(2018ZX097110010-001-012)。

摘　　要：目的 探究食欲刺激素(ghrelin)对乙酸诱导大鼠的保护作用及机制。方法 Wistar大鼠胃浆膜下微量注射30μL乙酸制备慢性胃损伤模型,假手术组仅打开腹腔后缝合。模型+食欲刺激素组大鼠ip给予食欲刺激素30μg·kg^(-1),每日2次。分别在连续给药第3,6和9天(D3,D6和D9)后,每组4只取胃测量溃疡面积;免疫组化法检测大鼠胃溃疡周围胃组织中增殖细胞核抗原(PCNA)阳性细胞比例,ELISA测定血清中白细胞介素1β(IL-1β)以及溃疡周围胃组织中IL-1β、肿瘤坏死因子α(TNF-α)和血管内皮生长因子(VEGF)的含量,NO合酶(NOS)试剂盒检测血清中诱导型NO合酶(iNOS)活性,Western印迹法测定胃溃疡周围胃组织中iNOS和内皮型NO合酶(eNOS)表达水平。结果 与假手术组相比,模型组大鼠D3胃展开可见穿孔样溃疡,D6胃溃疡创口面积未见明显缩小,D9溃疡创口面积开始缩小;与模型组相比,模型+食欲刺激素组D3大鼠胃溃疡处即形成新生基底组织,D6和D9胃溃疡创口面积显著缩小(P<0.05,P<0.01)。与假手术组相比,模型组大鼠D3溃疡周围胃组织中无PCNA阳性细胞,D6和D9呈增加趋势;与模型组相比,模型+食欲刺激素组大鼠D3胃溃疡周围组织中PCNA阳性细胞比例较高,D6和D9进一步增加。与假手术组相比,模型组大鼠血浆中IL-1β的含量显著增加(D3和D6,P<0.01),iNOS活性显著增加(D3,D6和D9,P<0.01),溃疡周围胃组织中IL-1β含量(D3和D6,P<0.01)和TNF-α含量(D3,P<0.01;D6,P<0.05)显著增加,iNOS蛋白表达显著增加(D3,D6和D9,P<0.01),VEGF含量显著降低(D3,D6和D9,P<0.01),而eNOS蛋白表达先降低(D3,P<0.05)后增加(D9,P<0.01);与模型组相比,模型+食欲刺激素组大鼠血浆中IL-1β含量显著降低(D3和D6,P<0.01),iNOS活性显著降低(D6和D9,P<0.01),溃疡周围胃组织中IL-1β和TNF-α含量均显著降低(D3和D6,P<0.01),iNOS蛋白表达显著降低(D3,P<0.01;D6,P<0.05;D9,P<0.01),VEGF含量显著增加(D3,D6和D9,P<0.01)eNOS蛋白表达先�OBJECTIVE To explore the protective effect and mechanism of appetite-stimulating hormone(ghrelin) on acetic acid-induced chronic gastric injury in rats. METHODS Wistar rats were microinjected with 30 μL of acetic acid subserosal to establish a model. In the sham operation group, the abdominal cavity of rats was opened and sutured without model establishment. The rats in the model+ghrelin group were given ghrelin 30 μg·kg;ip, twice per day. After continuous administration of 3, 6,and 9 d(D3, D6 and D9), 4 rats in each group were taken to measure the ulcer area. The immunohistochemical method was used to detect proliferating cell nuclear antigen(PCNA) positive cells in gastric tissue around gastric ulcers. The contents of interleukin-1β(IL-1β) in serum and IL-1β, tumor necrosis factor α(TNF-α) and vascular endothelial growth factor(VEGF) in gastric tissue around ulcers were determined by ELISA. The activity of inducible NO synthase(iNOS) in serum was detected by the enzyme kit, and the expression levels of iNOS and endothelial NO synthase(eNOS) in tissues around gastric ulcers were determined by Western blotting. RESULTS Compared with the sham operation group, perforated ulcers were seen in the gastric expansion of rats in the model group at D3. The areas of wounds of the gastric ulcers at D6 did not shrink significantly, but the area of ulcers at D9 began to shrink.Compared with the model group, new basal tissue was formed at the gastric ulcer site in the model+ghrelin group at D3, and the area of gastric ulcers was significantly reduced at D6 and D9(P<0.05, P<0.01). Compared with the sham operation group, there were few PCNA positive cells in the gastric tissue around ulcers in the model group at D3, but at D6 and D9 PCNA positive cells gradually increased. Compared with the model group, the content of PCNA positive cells in the tissues around gastric ulcers of rats was higher in the model+ghrelin group at D3, and at D6 and D9 PCNA positive cells kept increasing. Compared with the sham-operated group, the

关 键 词：食欲刺激素 胃溃疡 白细胞介素1Β 肿瘤坏死因子Α NO合酶 血管内皮生长因子 

分 类 号：R975[医药卫生—药品] R963[医药卫生—药学]