基于TLR4信号转导通路的补肾解毒方抗核辐射免疫损伤机制的研究

Nuclear radiation and leakage of radioactive substances generated from radiation facilities pose a grave threat to human health and life. Researches both domestic and overseas, in recent years, have shown that TLR4 signaling pathway is an important way for radiation immune injury and TLR4 ligand LPS may reduce radiation injury through TLR4 pathway. Previous studies indicated that nourishing yin and invigorating the kidney recipe, Liuwei Dihuang pill, has a better protective effect on nuclear radiation injury. According to clinical experience and experimental results, we optimize the prescription by adding heat-clearing and detoxicating Traditional Chinese Medicine at the foundation of supplementing nourishing yin and invigorating the kidney with the standard of syndrome differentiation, forming the Bushen Jiedu recipe. Our project intends to clarify the target and mechanism of TLR4 signaling pathway of Bushen Jiedu recipe on radiaction-protective effect. We experiment with TLR4 gene knock-out rats, use flow cytometry, ELISA, Real-time PCR, Western bolt and other techniques. We aim to observe the expression and the trends of TLR4 and downstream cytokines of the rats after radication, clarify the target and the mechanism in the sight of inflammatory immunity, so as to lay a foundation in theory and provide experimental evidence for radiaction-protective Traditional Chinese Medicine with low toxity, high efficiency and prevention and treatment integration.

核设施产生的核辐射及其泄漏的放射性物质严重威胁着人类的健康和生命。近年来国内外研究显示，TLR4信号转导通路是辐射免疫损伤的重要途径，并且TLR4配体LPS可通过TLR4通路减轻辐射损伤。我们的前期研究发现，滋阴补肾方六味地黄丸对核辐射损伤有较好的防护效果。根据既往临床经验和实验结果，我们将配方进行了优化组合，在滋阴补肾基础上辨证增加清热解毒的中药，形成在研的补肾解毒方。本项目拟从TLR4信号转导途径对补肾解毒方抗辐射的作用机制及靶点进行研究。在实验中采用TLR4基因敲除大鼠，应用流式细胞分析、ELISA、Real-time PCR、Western bolt杂交等方法，系统观察辐射后大鼠TLR4及其下游细胞因子动态含量及表达水平的变化，从炎症免疫的角度探讨该方的防护作用机制及靶点，为研制出低毒、高效、防治结合的核辐射中药防护剂奠定理论基础，提供科学的实验数据。

摘要：核能是解决未来能源供给问题的重要支柱，然而核设施泄露的放射性物质，严重威胁人类的健康与生命。辐射损伤导致的免疫功能障碍是急性放射性损伤患者最常见的并发症，至今仍是临床亟待解决的难题。近年来国内外研究显示，TLR4信号转导通路是辐射免疫损伤的重要途径，并且TLRT4配体LPS可通过TLR4通路减轻辐射损伤。本项目从TLR4信号转导途径对补肾解毒方抗辐射免疫损伤的作用机制及靶点进行初步探索。在实验中采用TLR4基因敲除小鼠，应用流式细胞分析、ELISA、Western bolt 杂交等方法开展研究，发现补肾解毒方可通过TLR4信号转导通路发挥辐射防护作用，能提高辐射小鼠存活率，增强胸腺、脾脏等组织修复能力，在提高机体免疫力的同时也有效抑制机体的自身炎性损伤，从而发挥辐射防护作用。