摘要
针对肝郁气滞型急性胰腺炎(AP)的中医病因病机和临床证候表现,构建病证结合大鼠模型,为相关中药复方药效学研究与功效网络验证提供评价工具。依照AP中、西医诊疗指南和肝郁气滞型AP的中医病因病机和临床证候表现,综合临床疗效Meta分析结果,结合实验室前期建立的病证结合动物模型构建方法与评价策略,抽提适宜于肝郁气滞型AP大鼠动物模型评价的生物学替代指标;分别运用慢性不可预知刺激(chronic unpredictable stress,CUS)法、慢性不可预知刺激+L-精氨酸(CUS+L-Arg)法构建大鼠模型,用上述生物学替代指标评价肝郁气滞型AP病证结合大鼠模型成立与否。实验期间观察记录大鼠体质量、证候积分,实验结束时手术留取大鼠血清、脏器、组织,检测各指标。与正常组相比,CUS组、CUS+L-Arg组大鼠证候积分均显著增高(P<0.01);反证药物柴胡舒肝丸可明显降低2组模型大鼠证候积分(P<0.01),表明2种造模方法均能复制肝郁气滞型AP大鼠模型中的肝郁气滞证候。与正常组相比,CUS+L-Arg组大鼠血清淀粉酶(AMY)活力水平升高3倍,具有统计学意义(P<0.01),而CUS组大鼠AMY活力水平升高,但尚不到正常值的3倍;与正常组相比,CUS+L-Arg组大鼠胰腺组织肿瘤坏死因子-α(TNF-α)、白细胞介素6(IL-6)mRNA水平显著升高(P<0.01),CUS组大鼠胰腺组织TNF-αmRNA水平显著升高(P<0.05),IL-6 mRNA水平仅具有升高趋势,表明仅有CUS+L-Arg法能够复制病证结合模型中的胰腺损伤。该研究提示CUS+L-Arg法连续造模4周,可建立肝郁气滞型AP大鼠病证结合模型,且模型具有可重复性、成模后稳定、动物死亡率低、与临床发病机制相近的特点,可用于中药药效评价与功效网络验证。
The aim of this paper was to construct a rat model of acute pancreatitis(AP)with syndrome of liver depression and Qi stagnation,and provide evaluation tools for pharmacodynamic research and efficacy network verification of related traditional Chinese medicine in view of the etiology,pathogenesis and clinical manifestations of AP.According to the Chinese and Western medicine diagnosis and treatment guidelines for AP with syndrome of liver depression and Qi stagnation,etiology,pathogenesis and clinical syndromes in TCM,Meta-analysis results,and evaluation strategy of establishing an animal model combining disease and syndrome in our laboratory,the biological surrogate outcomes suitable for the evaluation of animal models of AP with syndrome of liver depression and Qi stagnation were extracted then.The chronic unpredictable stress(CUS)method and chronic unpredictable stress+L-arginine(CUS+L-Arg)method were used to construct the rat model,and the above biological surrogate outcomes were used to evaluate whether an AP rat model was established.During the experiment,the weight and syndrome scores of the rats were observed and recorded.At the end of the experiment,the rats'serum,organs and tissues were collected from the operation to detect the various indicators.As compared with the normal group,the syndrome scores of the CUS group and CUS+L-Arg group were significantly increased(P<0.01);the anti-syndrome medicine Chaihu Shugan Pills could significantly reduce the syndrome scores of the two groups of rats(P<0.01),indicating that both modeling methods can replicate the syndrome of liver depression and Qi stagnation in the rat model.As compared with the normal group,the serum amylase(AMY)activity level was increased by 3 times in the CUS+L-Arg group(P<0.01),and the AMY activity level was also increased in CUS group,but not up to 3 times of the normal value.As compared with the normal group,the levels of TNF-α and IL-6 mRNA in the pancreatic tissues of the CUS+L-Arg group were significantly increased(P<0.01);the levels of TNF-α mRNA in the pancreatic tissues of the CUS group were significantly increased(P<0.05),but IL-6 mRNA level only showed a rising trend,indicating that only the CUS+L-Arg method can be used to replicate the AP damage in the disease-syndrome combination model.The CUS+L-Arg method can be used for continuous modeling for 4 weeks to establish a disease-syndrome combination model of AP rats with syndrome of liver depression and Qi stagnation.The model has the characteristics of repeatability,stability after modeling,low animal mortality,and similar clinical pathogenesis.It can be used for the evaluation of traditional Chinese medicine efficacy and the verification of efficacy network.
作者
孙凯滨
孙蓉
SUN Kai-bin;SUN Rong(Institute of Medical Sciences,the Second Hospital,Cheeloo College of Medicine,Shandong University,Ji'nan 250033,China;School of Pharmacy,Shandong University of Traditional Chinese Medicine,Ji'nan 250355,China;Advanced Medical Research Institute,Cheeloo College of Medicine,Shandong University,Ji'nan 250012,China)
出处
《中国中药杂志》
CAS
CSCD
北大核心
2020年第23期5732-5737,共6页
China Journal of Chinese Materia Medica
基金
山东省重点研发计划项目(2016ZDJS07A21,2017CXGC1301)
泰山学者工程专项经费项目(ts201511107)。
关键词
急性胰腺炎
肝郁气滞证
病证结合模型
慢性不可预知刺激
acute pancreatitis
syndrome of liver depression and Qi stagnation
disease-syndrome combination model
chronic unpredictable stress
作者简介
通信作者:孙蓉,博士,教授,博士生导师,主要从事中药药理与毒理研究,E-mail:sunrong107@163.com;孙凯滨,硕士研究生,E-mail:sunkaibin122@126.com。
