基于血瘀探讨化瘀通便汤调控PI3K/Akt/mTOR介导的慢传输型便秘 Cajal间质细胞自噬的机制

The reduction of interstitial cells of Cajal (ICCs) is one of the main mechanisms of slow transit constipation (STC), but it is not clear what mechanisms affect ICCs reduction. Excessive autophagy of ICCs can lead to a decrease in the number of ICCs, while PI3K/Akt/mTOR is one of the most important autophagy related signaling pathways in eukaryotic cells, and it is worth exploring how to regulate the PI3K/Akt/mTOR signaling pathway to regulate ICCs autophagy. Autophagy is closely related to blood stasis. Autophagy can lead to apoptosis of vascular endothelial cells and vascular wall injury, which is consistent with the pathogenesis of TCM blood stasis. STC patients have a long history in Clinic and many have blood stasis symptoms.Huayu Tongbian decoction has curative effect on STC，but the mechanism if it is by regulating the PI3K/Akt/mTOR signaling pathways intervention ICCs autophagy to treat is unknown. Previous studies have found that the number of ICCs in STC rats treated with Huayu Tongbian decoction is increased, therefore, it is speculated that the Huayu Tongbian decoction can control the autophagy of the ICCs by regulating the PI3K/Akt/mTOR signaling pathway to treat STC. This topic will treat STC rats with Huayu Tongbian decoction, and discuss the role of ICCs autophagy in STC and the partial mechanism of the treatment of the disease by blood stasis mechanism, and provide new ideas for TCM treatment of STC.

结肠Cajal间质细胞（ICCs）减少是慢传输型便秘（STC）发生的主要机制之一，但什么影响ICCs减少尚不明确。ICCs过度自噬可导致ICCs数量减少，而PI3K/Akt/mTOR是真核细胞最重要的自噬相关信号通路之一，调控PI3K/Akt/mTOR如何调控ICCs自噬值得探讨。自噬与血瘀关系密切，自噬失调可致血管内细胞凋亡，血管壁损伤，与中医血瘀的致病机制相符合。临床上STC患者病史长，多伴血瘀症状，化瘀通便汤治疗STC疗效确切，但是否通过调控PI3K/Akt/mTOR信号通路干预ICCs自噬而治疗的机制不详。前期研究发现，化瘀通便汤治疗的STC大鼠ICCs数量增加，因此推测化瘀通便汤可通过调控PI3K/Akt/mTOR调控ICCs自噬而治疗STC。本课题将对STC大鼠化瘀通便汤治疗，从血瘀机制探讨ICCs自噬在STC中的作用及化瘀通便汤治疗本病的部分机制，为中医治疗STC提供新思路。

结肠Cajal间质细胞（ICCs）减少是慢传输型便秘（STC）发生的主要机制之一，ICCs过度自噬可导致ICCs数量减少，PI3K/Akt/mTOR是真核细胞最重要的自噬相关信号通路之一，调控PI3K/Akt/mTOR干预ICCs自噬值得探讨。制备慢传输型便秘大鼠模型，化瘀通便汤干预治疗，观察各组大鼠体重及一般状态，检测各组大鼠ICCs超微结构及数量变化，检测c-Kit、HCN1、HCN2蛋白表达水平，检测化瘀通便汤对PI3K/Akt/mTOR信号通路上自噬相关蛋白Beclin1、LC3B、Atg5、p62表达的影响。结果表明，化瘀通便汤可改善慢传输型便秘大鼠毛发暗沉、暴躁易怒状态，恢复体重，改善和修复ICCs超微机构，使得Cajal间质细胞数量显著增加，促进c-Kit、HCN1、HCN2蛋白表达能力的提升，主要机制可能与PI3K/Akt/mTOR信号通路上，下调Beclin1、LC3B、Atg5表达而抑制ICCs过度自噬，上调p62表达促进ICCs修复有关。本项目研究表明活血化瘀中药能够干预ICCs自噬，一定程度恢复肠道蠕动，促进排便，为中医治疗STC提供新思路。