李甫 陈萌 吉建梅 张晞文 丁俊 黄金鑫 唐睿 蔡淦

摘要 基于芍藥甘草汤具有显著改善Oddi括约肌功能障碍（SOD）患者腹痛的临床疗效和抑制自噬相关蛋白（GPR78、Beclin1）的工作基础，结合Cajal间质细胞（ICC）过度自噬将引起平滑肌收缩紊乱、进一步导致SOD的研究进展，探讨芍药甘草汤调节ICC自噬水平及治疗SOD的分子药理机制。首先，复制豚鼠SOD模型，观察芍药甘草汤对Oddi括约肌压力、ICC自噬水平的影响;其次，吗啡刺激诱导离体SOD模型，观察芍药甘草汤对Oddi括约肌肌条收缩曲线、自噬通路的影响;最后，分离原代ICC、构建IRE1基因沉默或过表达载体、衣霉素刺激ICC建立内质网应激模型，予不同浓度的芍药甘草汤孵育，观察ICC内质网应激水平、自噬信号通路的变化。最终验证“芍药甘草汤抑制IRE1/Beclin1信号通路，减轻ICC内质网应激引起的过度自噬，从而改善SOD平滑肌舒张效应”的科学假说，为推进中医药调节胆道动力学的理论研究夯实基础。

关键词 芍药甘草汤;Oddi括约肌功能障碍;Cajal间质细胞;细胞自噬;IRE1/Beclin1通路

Study on the Mechanism of Shaoyao Gancao Decoction on Improving Sphincter Dysfunction

of Oddi by Regulating Autophagy of Interstitial Cells of Cajal

LI Fu， Chen Meng， JI Jianmei， ZHANG Xiwen， DING Jun， HUANG Jinxin， TANG Rui， CAI Gan

（Shuguang Hospital Affiliated to Shanghai University of Traditional Chinese Medicine， Shanghai 201203， China）

Abstract Based on the clinical effect of Shaoyao Gancao Decoction on abdominal pain of Sphincter of Oddi dysfunction （SOD） patients and the work basis of inhibiting autophagy related proteins （GPR78，Beclin1）， combined with the research progress that Cajal interstitial cells （ICC） over autophagy will cause smooth muscle contraction disorder and further lead to SOD research development， to explore the molecular pharmacological mechanism of Shaoyao Gancao Decoction on regulating the level of ICC autophagy and treating SOD. Firstly， the SOD model of guinea pig was duplicated， and the effects of Shaoyao Gancao Decoction on Oddi sphincter pressure and ICC autophagy level was observed; secondly， morphine stimulation induced SOD model in vitro， and the effects of Shaoyao Gancao Decoction on Oddi sphincter contraction curve and autophagy signal pathway of Oddi was observed; finally， we isolated the primary ICC， constructed the silencing or overexpression vector of IRE1 gene， and stimulated ICC by tunicamycin to establish the endoplasmic reticulum stress model， and incubated with different concentrations of Shaoyao Gancao Decoction to observe the changes of endoplasmic reticulum stress level and autophagy signal pathway in ICC. We verified the scientific hypothesis that Shaoyao Gancao Decoction can inhibit IRE1 / Beclin1 signal pathway， reduce excessive autophagy caused by ICC endoplasmic reticulum stress， so as to improve the relaxation effect of SOD smooth muscle， which will lay a solid foundation for the theoretical research of TCM regulating biliary dynamics.

Keywords Shaoyao Gancao Decoction; Oddi Sphincter dysfunction; Interstitial cells of Cajal; Cell Autophagy; IRE1/Beclin1 pathway

中图分类号：R259文献标识码：Adoi：10.3969/j.issn.1673-7202.2021.02.025

Oddi括约肌（Sphincter of Oddi，SO）是胆、胰与十二指肠连接部位的精密平滑肌装置，在神经、体液、局部反射的多重复杂调控下，形成时相性的移行复合运动（MMC），发挥控制胆汁和胰液排放、防止反流的“门户”作用[1]。Oddi括约肌功能障碍（Sphincter of Oddi Dysfunction，SOD）是因SO舒张功能异常引起的一组疾病，临床分为胆型和胰型2种，以胆型SOD常见，主要表现为上腹部疼痛发作30 min以上、足以影响日常活动甚至到急诊科就诊，排便或改变体位、服用抗酸药均不能缓解，可伴随恶心、呕吐、肝功能异常、胆管扩张、胆管排空时间延长等表现[2]（罗马Ⅳ标准，2106）。……