胃溃疡肝郁证候大鼠模型的建立及其评价

胃溃疡肝郁证候大鼠模型的建立及其评价

目的探讨胃溃疡肝郁证候动物模型的建立及电针的治疗作用。方法采用Open-field法选择60只SD大鼠，适应性喂养7 d，随机分为空白组、模型组、模型对照组和电针组，每组15只。除空白组外，其余3组均采用慢性不可预见性刺激合并醋酸烧灼法建立胃溃疡抑郁证大鼠模型。模型对照组继续慢性不可预见性刺激，电针组电针“肝俞”“梁丘”治疗，连续13 d。观察大鼠一般状态、旷场实验结果、溃疡指数，RT-PCR检测海马组织脑源性神经营养因子（BDNF），ELISA 检测血清胃泌素（GAS）含量。结果实验第21日，除空白组外其余3组大鼠出现精神萎靡、四肢无力、被毛乏泽、食欲降低等表现。实验第34日，模型对照组大鼠一般状况表现较模型组严重，电针组大鼠一般状况明显改善，模型组大鼠水平和垂直穿格数较空白组显著减少（P＜0.01），与模型组比较，模型对照组大鼠水平和垂直穿格数显著减少（P＜0.05），电针组大鼠水平和垂直穿格数显著增加（P＜0.01）；模型组大鼠溃疡指数较空白组显著升高（P＜0.01），与模型组比较，模型对照组大鼠溃疡指数显著升高（P＜0.05），电针组大鼠溃疡指数显著下降（P＜0.01）；模型组海马组织BDNF mRNA表达较空白组显著下降（P＜0.01），与模型组比较，模型对照组海马组织BDNF mRNA表达显著下降（P＜0.05），电针组海马组织BDNF mRNA表达组显著升高（P＜0.01）；模型组大鼠GAS含量较空白组显著升高（P＜0.01），与模型组比较，模型对照组大鼠GAS含量显著升高（P＜0.05），电针组大鼠GAS含量显著降低（P＜0.05）。结论通过复合方式建立病证结合胃溃疡肝郁证候动物模型可行，电针“肝俞”和“梁丘”对该模型有治疗作用。

Abstract：Objective To discuss the establishment of gastric ulcer liver depression syndrome rat model and the therapeutical effects of electroacupuncture. Methods As the result of Open-field experimental method，60 SPF male SD rats were selected as experimental animals. After 7 days of adaptive feeding，60 rats were randomly divided into blank group，model group，model control group，and electoracupuncture group，15 rats in each group. Except for the blank group，the other three groups were stimulated by chronic unpredictable stimulation to establish rat depression model. Then model control group was once again stimulated by chronic unpredictable stimulation for 13 days；electoracupuncture group was treated at Gan Shu （BL18）and Liang Qiu （ST34）acupoints for 13 days. General state of rats，Open-field experimental results and gastric ulcer index were observed. BDNF was measured by RT-PCR. GAS content was measured by ELISA. Results After modeling for 21 days，except for the blank group，the other three

基金項目：国家自然科学基金（81373718）；辽宁省自然科学基金（2013020179）；辽宁省教育厅科学研究项目（L2012339）；辽宁省科技攻关项目（2009225010-36）；沈阳市高新技术产业发展与科技攻关计划（F12-193-9-37）；沈阳市科技局应用基础研究专项（F13-318-1-67）

groups appeared listlessness，limb weakness，and fur dry and lose luster，

appetites reduced，etc. After modeling for 34 days，compared with model group，model control group had more severe general state than model group. After acupuncture treatment，the general state of acupuncture group was improved. The horizontal and vertical activity sore in model group decreased significantly compared with the blank group （P＜0.01）. Compared with the model group，the horizontal and vertical activity sore in model control group decreased significantly （P＜0.05），and increased significantly in the electroacupuncture group （P＜0.01）. Ulcer index in the model group increased significantly compared with the blank group （P＜0.01）. Ulcer index in the model control group increased significantly compared with the model group （P＜0.01）. Ulcer index in electroacupuncture group decreased significantly （P＜0.01）. BDNF mRNA expression in hippocampus of model group decreased significantly compared with the blank group （P＜0.01）. Compared with model group，the BDNF mRNA expression in hippocampus of model control group decreased significantly （P＜0.05）. The BDNF mRNA expression in the electroacupuncture group increased significantly （P＜0.01）. The GAS content in the model group increased significantly compared with the blank group （P＜0.01）. Compared with the model group，the GAS content in the model control group increased significantly （P＜0.05）. The GAS content in the electroacupuncture group decreased significantly （P＜0.05）. Conclusion The establishment of depressive rat gastric ulcer model which combining disease and syndrome together proves the feasibility of this animal experimental model. Electroacupuncture for Gan Shu （BL18）and Liang Qiu （ST34）can improve the efficacy of this model.Key words：gastric ulcer；depression；animal models；electroacupuncture；rats

随着生物医学模式向社会-心理-生物医学模式转变，越来越多的疾病受到情感情绪变化的影响。中医学认为，情志不遂、忧思恼怒、肝气郁结可致胃脘疼痛、纳呆、胀满。清代叶天士提出“肝为起病之源，胃为传病之所”，强调情志因素在脾胃病中的重要作用。有研究显示，消化性溃疡和功能性胃肠疾病的发生与抑郁情绪密切相关[1-2]。本实验依据上述理论，探讨胃溃疡肝郁证候模型的建立及电针的治疗作用，为建立病证结合的动物模型提供参考。

1 实验材料

1.1 动物

SPF级雄性SD大鼠60只，2月龄，体质量140～200 g，辽宁中医药大学实验动物中心，动物合格证号SCXK（辽）2010-0001。饲养于室温18～23 ℃、相对湿度35%环境，每日光照时间约为11 h。

1.2 主要试剂与仪器

10%水合氯醛（青岛宇龙海藻有限公司），DAB显色试剂盒（北京博奥森生物科技有限公司），脑源性神经营养因子（BDNF）扩增产物（北京三博远志生物科技有限公司），DL2000 DNA marker（北京全氏金生物科技有限公司），胃泌