青黛、参三七调控NLRP3炎症小体/Smad信号通路介导溃疡性结肠炎EMT的 机制研究

Ulcerative Colitis (Ulcerative Colitis,UC) is a disease with the features of prolonged disease course and easily relapse, whose morbidity rate is rising year by year. UC is extremely hard to cure and has canceration risk, while Traditional Chinese Medicine(T.C.M.) has incomparable advantage in treating it. Qingchang Suppository is developed by Professor Ma Guitong, a renowned T.C.M. physician in Shanghai. Its efficacy has been demonstrated in our 18 years' clinical practice and studies, among which a pre-clinical study has shown that Qingdai and Sanqi, the major constituent of Qingchang Suppository, improved enteric permeability. In this subject, we intend to investigate the mechanisms of Qingdai and Sanqi on treating UC by means of inhibiting intestinal epithelial-mesenchymal transition（EMT）from the perspective of regulating NLRP3 inflammasome/ Smad pathway, thus strengthening the intestinal barrier function, repairing the intestinal tissue, inhibiting intestinal fibrosis and preventing cancerization. We intend to prove the hypothesis that NLRP3 inflammasome could up-regulating TGF-β1 and activate Smad pathway to regulate the expression of Snail which may lead to the intestinal epithelial-mesenchymal transition. To further study the inner link between intestinal EMT and Heat-Resolving and Detoxication and promoting blood circulation for removing blood stasis in T.C.M. In order to further improve clinical theory of T.C.M in UC treatment and demonstrate T.C.M advantage in UC treatment.

溃疡性结肠炎（UC）患病率逐年上升，病程长，反复发作，治愈难度大，且有癌变风险，中医药治疗本病具有优势。清肠栓由上海市名中医马贵同教授研发，临床运用三十余年，疗效肯定。前期研究证明清肠栓及其主药青黛、参三七可改善肠道通透性，本课题以葡聚糖硫酸钠诱导溃疡性结肠炎大鼠和大鼠结肠上皮细胞为研究对象，从调控NLRP3炎症小体/Smad信号通路的角度，研究清肠栓的主药青黛、参三七通过抑制肠上皮间质转化从而改善肠道通透性、维持肠道稳态、抗肠道纤维化以及防止UC癌变的作用机制；验证NLRP3炎症小体通过上调 TGF-β1，激活Smad通路，调控Snail基因表达，从而诱发肠道上皮间质转化（EMT）的假说；进一步阐述“肠上皮间质转化”与“热毒瘀滞”内在联系，探讨中医学清热解毒、活血化瘀治疗的生物学特性，为清热解毒化瘀治疗UC提供现代生物学证据，进一步完善中医药治疗UC治疗中的作用和优势。

溃疡性结肠炎（Ulcerative Colitis, UC）患病率逐年上升，病程长，反复发作，治愈难度大，且有癌变风险，中医药治疗本病具有优势。清肠栓由上海市名中医马贵同教授研发，临床运用三十余年，疗效肯定。前期研究证明清肠栓及其主药青黛、参三七可改善肠道通透性，本研究进一步在体内实验和体外实验水平，探讨清肠栓、清肠栓主药（青黛、参三七）从调控NLRP3炎症小体/Smad信号通路的角度，研究清肠栓的主药青黛、参三七通过抑制NLRP3炎症小体活化，调控TGF-β/Smad信号通路，从而调控肠道黏膜屏障功能，维持肠道稳态的作用机制。通过研究，我们发现：1、在体内实验中，清肠栓、清肠栓主药（青黛、参三七）能有效抑制结肠组织中NLRP3炎症小体活化，调控NLRP3、ASC、Caspase-1及其下游炎症因子IL-1β、IL-18的表达。2、青黛、三七能有效调控结肠组织中TGF-β/Smad信号通路相关TGF-β、p-Smad2/3、Smad2/3、Smad4的表达。3、对青黛、三七能调控上皮间质转化进行探索性研究，青黛、三七干预后一定程度上缓解结肠组织中E-cadherin的表达下调趋势。4、通过NLRP3-/-小鼠进一步证实了NLRP3炎症小体为调控TGF-β/Smad信号通路的重要调控因素。5、在体外实验中，青黛、参三七的有效提取物靛玉红、人参皂苷Rg1能有效抑制NLRP3炎症小体活化，调控NLRP3、ASC、Caspase-1及其下游炎症因子IL-1β、IL-18的表达，从而调控TGF-β/Smad信号通路。6、本课题从现代生物学角度揭示UC“热毒瘀滞”及清热解毒，活血化瘀中药治疗的生物学特性。分析“NLRP3炎症小体”与“热毒瘀滞”内在联系，讨论TGF-β/Smad信号通路与痢疾（UC）发生发展之间的关系，初步探讨UC“热毒瘀滞”与EMT发生的可能，以及中药青黛、三七清热解毒、活血化瘀治疗的生物学特性。为清热解毒化瘀治疗UC提供现代生物学证据，进一步完善中医药治疗UC的临床理论依据，从而牢固确立其在UC治疗中的作用和优势。