茵陈蒿汤治疗重症急性胰腺炎长链非编码RNA DLEU2/miR-30a-5p信号轴异常的机制诠释“古方新用”

Severe acute pancreatitis (SAP) is one of the common acute abdomen diseases with high morbidity and mortality. The pathogenesis of AP is not yet completely clear, and there is still a lack of effective and specific treatments. Our preliminary study found that Yinchenhao Decoction can induce apoptosis of damaged acinar cells, reduce inflammation, and has a good therapeutic effect on AP, which is worth further investigation and exploration. In addition, we found that miR-30a-5p may inhibit the activation of inflammatory signaling pathways in AP rats. In certain tumor cells, miRNA-30a-5p negatively regulates the expression of Beclin-1, BCL-2 and XIAP, and promotes the inhibition of autophagy and the promotion of apoptosis. LncRNA DLEU2 down-regulates miR-30-5p expression and relieves its inhibitory effect on downstream target genes. This project plans to use in vitro and in vivo models and clinical patients as research objects on the basis of the previous period, taking the signal axis of lnc RNA DLEU2/miR-30a-5p as the cut-in to further investigate the signal transduction network of Yinchenhao Decoction through regulating cell death and inflammation, and the molecular mechanism of the multi-level, multi-pathway prevention and treatment of AP. We also design to interpret the modern biological basis for the treatment of traditional Chinese medicine based on the new application of old medicine, providing an effective strategy for the therapy of AP.

重症急性胰腺炎（SAP）是临床急危重症，病死率高，治疗棘手，我们前期研究发现茵陈蒿汤能诱导受损的胰腺腺泡细胞凋亡，减轻炎症反应，对SAP具有治疗作用；其组分大黄的提取物大黄素能通过上调miR-30a-5p抑制SAP大鼠模型炎症信号通路的激活。在某些肿瘤细胞中，miRNA-30a-5p能抑制自噬、促进凋亡，LncRNA DLEU2可下调miR-30a-5p。本研究在前述基础上，以细胞-动物-临床病例为研究对象，通过体外细胞实验-动物在体实验-临床病例观察，以lncRNA DLEU2靶向调控miR-30a-5p为切入点，采用基因敲除、体内外转染及siRNA干扰等技术，深入研究SAP胰腺腺泡细胞lncRNA DLEU2/miR-30a-5p信号轴对腺泡细胞自噬-凋亡-炎症级联信号转导网络的影响，以及茵陈蒿汤的作用靶点。阐明SAP部分发病机制，诠释茵陈蒿汤“古方新用”治疗SAP的现代生物学机理。

重症急性胰腺炎（SAP）是临床急危重症，病死率高，治疗棘手。我们前期研究发现茵陈蒿汤及其主要成分大黄素能减轻SAP炎症反应。本研究在前述基础上，以细胞-动物为主要研究对象，深入探讨SAP病程中胰腺腺泡细胞自噬-凋亡-炎症级联反应的信号网络以及茵陈蒿汤的潜在干预途径。在牛磺胆酸钠（STC）诱导的SAP体内、外模型中证明存在miR-30a-5p及其下游靶蛋白表达差异，miR-30a-5p表达水平下调，XIAP，Beclin-1，LC-3II和NF-κB蛋白表达水平上调，miR-30a-5p通过调控胰腺腺泡细胞自噬-凋亡-炎性级联反应转导网络，在SAP发生发展中发挥重要作用。LncRNA-PVT1靶向miRNA-30a-5p调控自噬-凋亡-炎症级联信号，促进凋亡坏死转换，恢复自噬通量。茵陈蒿汤可以减轻SAP大鼠胰腺损伤和炎症反应，诱导胰腺细胞凋亡，抑制自噬反应。LncRNA-PVT1/miR-30a-5p介导的细胞自噬和凋亡可能是茵陈蒿汤治疗SAP的关键药物靶点之一。建立了茵陈蒿汤指纹图谱，通过表征茵陈蒿汤的主要化学成分，建立了茵陈蒿汤质控标准。同时，基于自主搭建的整合组学平台和临床样本，进行了探索性研究，发现了血清极性小分子可作为早期预警SAP的生物标记物。我们的研究有利于阐明SAP部分发病机制，为进一步开发中药提供实验依据。该项目发表SCI论文4篇，中文核心期刊论文2篇，申报发明专利3项，授权2项，获得科研奖励1项，培养博硕士研究生5人。