呕必宁对顺铂诱发脑及胃组织SP及NK1受体改变的影响(1)
摘 要：目的：观察呕必宁对顺铂诱发大鼠胃、延髓组织SP含量及NK1受体表达改变的影响，探讨呕必宁防治顺铂诱发呕吐机制同时，为中医学“胃气上逆”等理论提供实验依据。方法：Wistar 大鼠 54只，分成对照组、模型组和中药组，分别灌服盐水、呕必宁煎液6天时，对照组腹腔注射盐水，余两组注射顺铂48h后，取延髓、胃组织，放免法检测SP含量，免疫组化法检测NK1表达，并进行灰度值测定。结果：模型组大鼠胃窦及延髓组织SP含量、NK1表达高于对照组和中药组(P﹤0.01)；中药组SP含量及NK1表达与对照组无明显差异(P﹥0.05)。结论：呕必宁能够抑制顺铂所致大鼠胃、延髓组织SP升高及NK1表达。大鼠胃及延髓组织SP增多、NK1表达上调可能是“胃气上逆”的病理学基础之一。
关键词：顺铂；SP；NK1受体；呕吐；呕必宁
中图分类号：R285.5
文献标识码：A
文章编号：1673-7717(2008)03-0466-03
Effect of Oubining on the Change of SP and NK1-R Induced by Cisplatin
WANG De-shan，LIU You-mei，MING Cai-rong，SHAN De-hong
(Liaoning University of Traditional Chinese Medicine，Shenyang 110032，Liaoning，China)
Abstract：Objective：In order to detect the mechanism of Oubining preventing and treating vomiting induced by cisplatin (DDP) and provide experimental basis for the "adverse ascending of Stomach-Qi"， action of Oubining to the changes of substance P (SP) concentration and neurokinin receptor 1 (NK1-R) in rats' stomach and medullar tissues induced by DDP. Methods：54 Wistar rats were randomly divided into three groups：control group， model group and Chinese herbal group， and the rats in later groups were given saline and oubining via intragastric administration. 6 days later， the rats in control group were given DDP and rats in other groups were given saline via intraperitoneal injection. 48 hours later medullar tissue and gastric antrum were taken out to measure SP levels with radioimmunoassay and detect NK1-R expression by immunohistochemistry technique. Results：SP concentration and NK1-R expression in the tissues from gastric antrum and medulla oblongata in the model group were higher than those in the control group and Chinese herbal group (P1-R expression between the Chinese herbal group and the control group (P>0.05). Conclusion：Oubining can inhibit the SP level increase and strong expression of NK1-R in medulla oblongata and gastric antrum induced by DDP. And the SP level increase and strong expression of NK1-R in medulla oblongata and gastric antrum might be the pathologic basis of “adverse ascending of Stomach-Qi”.
Keywords：Cisplatin (DDP)；SP；NK1-R；vomiting；Oubining
临床及实验研究证明呕必宁具有良好的防治化疗药诱发呕吐的效果，其作用机制与体内胺类及神经肽类等物质改变有关[1-2]。近年研究发现 P物质(SP)也是致吐的内源性配体之一，速激肽受体(NK1)拮抗剂无论对急性还是延迟性呕吐均有良好的疗效[3-4]。为了进一步探讨呕必宁防治顺铂(DDP)诱发呕吐作用机制，进行了以下研究工作。
1 材料与方法
1.1 实验动物
Wistar健康成年大鼠54只，体重(190± 20)g，雌雄不限。由辽宁中医药大学实验动物中心提供，动物许可证号：SCXK(辽)2004-0018。
1.2 呕必宁
由人参、竹茹、橘皮、大枣、半夏、甘草等中药组成，购于辽宁中医药大学附属医院，药物真伪性由辽宁中医药大学中药鉴定室进行鉴定。按传统方法煎煮，浓缩成0.5g/mL生药含量。0～4℃冰箱保存，灌胃前加热。
1.3 顺铂
齐鲁制药有限公司生产，国药准字H20023460，产品批号：05030082。
1.4 免疫组化试剂
兔抗鼠SP受体抗体、即用型SABC免疫组化染色试剂盒及DAB染色试剂均购自武汉博士德生物工程有限公司；SP放免试剂盒购于北京华英生物技术研究所；BI2000图像分析系统：成都泰盟科技有限公司生产。
1.5 实验方法
1.5.1动物分组与给药 编号后按体重采用随机数字表分为3组，每组18只；①对照组(下称：NS组)：生理盐水灌胃早、晚各1次，剂量0.7mL·kg-1体重。于标本采集前48h，一次性经腹腔注射生理盐水1mL·kg-1体重。②顺铂+盐水组(下称：模型组)：生理盐水灌胃，用法、剂量同上，于标本采集前48h，一次性经腹腔注射DDP，用量 10mg·kg-1体重。③顺铂+中药组(下称：中药组)：呕必宁灌胃，剂量0.7mL·kg-1体重，早晚各1次，于标本采集前48h，注射DDP，方法、剂量同模型组。(王德山 刘永梅 明彩荣 单德红)