Cajal间质细胞在慢性结石性胆囊炎胆囊动力减弱机制中的作用及茵陈蒿汤的干预机制

The gallbladder is the engine for power of biliary motility. Weakening motility of gallbladder is a main part of the pathogenesis for cholecystitis, cholelithiasis and other biliary diseases.Traditional Chinese medical science believes that chronic lithous cholecystitis belongs to the category of "gallbladder distention", "hypochondriac pain" and "jaundice". The pathogenesis is dampness-heat of liver and gallbladder, liver depression and qi stagnation (GanDanShiRe and GanYuQiZhi). The treatment is soothing Liver Cholaneresis, compared with clear heat and eliminate dampness (QingReChuShi and ShuGanLiDan ). Applying YinChenHao decoction to clear heat and eliminate dampness is proved to be the representative prescription. It has been proved that, without jaundice, the effect of Yinchenhao Decoction to dampness-heat of liver and gallbladder is more significant;also the effect of YinChenHao Decoction in the treatment of lithous cholecystitis is more significant, It can enhance the contraction function of gallbladder, but the exact mechanism is still not clear. . Studies have shown that the interstitial cells of Cajal(ICC) of gallbladder possibly lead to gallbladder spontaneous rhythmic movement and adjustment of smooth muscle contraction, thus playing the same role as ICC in gastrointestinal duct in gallbladder motility disorders. In our subject, we will study the gallbladder of cholesterol gallstone model of guinea pig and the gallbladder of chronic lithous cholecystitis of human, trying to elaborate the function and mechanism of ICC in weakening cholecystic contraction, and to explore the intervention mechanism to ICC by the traditional Chinese medicine Yinchenhao Decoction , the serum of YinChenHao decoction and its effective ingredients through means of cell biology, laser copolymerization of electron microscopy, molecular biology, electrophysiology (patch clamp, etc.) and other advanced methods, thus we can provide related theoretical grounding for treating gallbladder motility weakening disease for Integrative Medicine.

胆囊是胆道动力的启动器官.胆囊动力减弱是胆囊炎、胆石症等疾病发病机制的主导环节。慢性结石性胆囊炎属中医"胆胀"、"胁痛"范畴。病机为肝胆湿热，肝郁气滞，治则清热除湿，疏肝利胆。茵陈蒿汤为清热除湿主治阳黄的代表方剂,实践证明，无黄疸而有肝胆湿热者，用之效果更加明显；茵陈蒿汤治疗结石性胆囊炎疗效显著，可增强胆囊收缩功能，确切机理还不清楚。研究证明：Cajal间质细胞(Interstitial cells of Cajal ICC )可能介导胆囊自发性节律的发生和平滑肌收缩活动的调节，在胆囊动力性疾病中发挥了与胃肠道动力障碍中相同的作用。本课题以豚鼠胆固醇结石模型和人慢性结石性胆囊炎胆囊为研究对象，通过细胞生物学、激光共聚焦显微镜、分子生物学和电生理（膜片钳等）等方法研究ICC在胆囊收缩减弱中的确切作用，探讨中药茵陈蒿汤对胆囊ICC干预作用机理,为中西医结合治疗胆囊动力障碍性疾病提供理论依据。

胆囊是胆道系统的重要组成部分。慢性结石性胆囊炎（Chronic Calculous Cholecystitis, CCA）是临床上多见的胆道系统疾病，其中胆道动力的减弱在其发病机制中起着重要作用，但是尚未有研究完全阐明胆道动力学改变的机制。本课题是前期研究的深入，以豚鼠胆固醇结石模型胆囊和人结石性胆囊炎胆囊为研究对象，在本课题的研究过程中我们采用western-blot法和RT-PCR法分和双重标记激光共聚焦检测别检测胆囊组织内Cx43、Cx45、C-kit蛋白、mRNA的表达以及，观察其变化。通过激光共聚焦显微镜测定细胞内Ca2+ 浓度，IP3 试剂盒测定胆囊ICC细胞三磷酸肌醇（IP3）及其受体表达的变化，明确中药调节胆囊ICC信号传导机制。研究采用用中药含药血清及其有效成分6，7-二甲氧基香豆素、大黄素和栀子苷分别处理胆囊ICC细胞，通过激光共聚焦显微镜以及膜片钳技术，观察内钙离子的变化、胆囊ICC细胞三磷酸肌醇（IP3）及其受体表达的变化及相关离子通道的变化，揭示ICC在胆囊动力减弱中的作用及机理；中药茵陈蒿汤、含药血清及其有效成分6，7-二甲氧基香豆素、大黄素和栀子苷对胆囊动力减弱干预作用机理：激活ICC的起搏电流、促进信号传导保护和维持ICC的正常形态和结构，修复神经-ICC-平滑肌网络的损伤，有效地改善胆囊运动障碍，增强胆囊收缩功能。本研究从细胞信息学和离子通道学的角度深入探讨了ICC在慢性结石性胆囊炎胆囊动力减弱中的作用，以及茵陈蒿汤改善胆囊运动功能的机制，为茵陈蒿汤的临床应用提供理论依据。