基于lncRNAMALAT1-ERK/p38MAPK-AQP-4信号轴探讨陇中消肿止痛合剂通过干预“血-脊髓屏障”防治继发性脊髓损伤的分子机制

In recent years, syndrome differentiation and treatment of traditional Chinese medicine （TCM）have shown some advantages in the treatment of secondary spinal cord injury (SSCI). So we make "fill the Du meridian, Qi and blood" as the principle, using the hospital preparation - Longzhong Xiaozhongzhitong Mixture that is hospital preparation in Gansu provincial hospital of TCM in the treatment of SSCI, has good clinical curative effect. At the same time, the protective effect of it on the spinal cord in SSCI was confirmed by previous experiments. Based on previous studies, we will further explore the protective effect of Longzhong Xiaozhongzhitong Mixture on blood-spinal cord barrier (BSCB) in SSCI and related molecular biological mechanisms through the establishment of the rat model of spinal cord injury and the cell model of hypoxia reoxygenation BSCB, in-depth analysis of the mechanism of lncRNAMALAT1 regulating the expression of AQP-4 in BSCB through ERK/p38MAPK signaling pathway from the level of gene regulation. It will clarify the intervention efficacy of Longzhong Xiaozhongzhitong Mixture on lncRNAMALAT1-ERK/p38MAPK-AQP-4 signal axis mechanism, provide a scientific basis for "fill the Du meridian, Qi and blood" of TCM in the treatment of SSCI, lay a theoretical foundation for the further popularization and application of the prevention and treatment of SSCI in Longzhong Xiaozhongzhitong Mixture, and provide a new strategy and target for the prevention and treatment of SSCI in TCM.

近年来，中医药辨证施治在治疗继发性脊髓损伤（SSCI）中体现出一定优势，为此我们以“补督脉、行气血”为法则，运用甘肃省中医院院内制剂-陇中消肿止痛合剂治疗SSCI，取得较好的临床疗效，并通过前期实验研究，证实其在SSCI中对脊髓的保护作用。本项目拟在前期研究基础上，进一步探讨陇中消肿止痛合剂对SSCI中血-脊髓屏障(BSCB)的保护作用及相关分子生物学机制，通过构建大鼠脊髓损伤模型及缺氧-复氧血-脊髓屏障细胞模型，从基因调控水平深入分析lncRNAMALAT1通过ERK/p38MAPK信号通路调控BSCB中AQP-4表达的机制，并阐明陇中消肿止痛合剂干预lncRNAMALAT1-ERK/p38MAPK-AQP-4信号轴的药效学机理。为中医“补督脉、行气血”法治疗SSCI提供科学依据，为陇中消肿止痛合剂防治SSCI的进一步推广应用奠定理论基础，为中医药防治SSCI提供新的策略思路及治疗靶点。

中医辨证施治在治疗早期继发性脊髓损伤中体现出一定优势，为此，我们以“补督脉、行气血”为法则，运用甘肃省中医院院内制剂-陇中消肿止痛合剂治疗，取得较好的临床疗效，并通过前期实验研究，证实了其在脊髓损伤中的保护作用。本研究以中医理论为基础，以现代医学理论为载体，揭示中医药防治脊髓损伤的现代科学本质，ERK/MAPKs信号通路的调节、AQP-4的表达与血-脊髓屏障损伤后的水代谢及神经修复密切相关，但具体基因调控机制尚不明确。目前中国人群中虽有通过通过中医辨证论治防治脊髓损伤的相关研究，但由于对中医病因病机认识不同，治疗侧重点不同，未形成统一的、行业内认可的辩证标准，阻碍了中医药防治本病的进一步发展，影响了中医药结合中医辨证论治理论优势的发挥和学术交流。故本研究以前期实验研究为基础，以中医“补督脉、行气血”为原则，首先通过网络药理学筛选陇中消肿止痛合剂中的活性化合物及对应靶标，探讨陇中消肿止痛合剂治疗脊髓损伤的作用机制，并通过建立大鼠脊髓损伤模型、大鼠缺氧-复氧血脊髓屏障体外实验模型，采用免疫荧光、蛋白质免疫印迹（WB）、逆转录-聚合酶链反应(RT-PCR)等实验方法验证经网络药理学挖掘出的lncRNA MALAT1- ERK/p38MAPK-AQP4信号轴分子生物学机制，阐明相关药效学机机理，进一步明确陇中消肿止痛合剂对脊髓损伤后血-脊髓屏障的保护作用（机制），并深入探讨lncRNAMALAT1-ERK/p38MAPK-AQP-4轴对血-脊髓屏障损伤后水代谢异常的分子调控机制，较全面的阐述陇中消肿止痛合剂治疗SCI的作用机制。为脊髓损伤后神经修复及功能重建提供新的研究靶点。本研究将有助于中医临床医疗工作者重视中医药辩证在脊髓损伤发生与发展中的重要作用，从基因调控水平进一步分析及认识脊髓损伤的发病机制，以期发挥中医药防治脊柱疑难病症方面的独特优势，并将为中医骨伤事业的发展带来一定的社会效益和经济效益。