藏药砂生槐种子水溶性生物碱E2-a抗泡球蚴病的分子机制研究

Echinococcosis has seriously affected the health of the people in the western regions and limited the local economic developments as well as social stability, and so far the effective and safe antiechinococcosis medicines have not been announced. Developing drugs that can antagonize the proliferation of cysts and induce host immunity is obviously one of strategies of echinococcosis control. Sophora moorcroftiana seed is a unique folk medicine for the treatment of insect poison in China. It has been found that water soluble alkaloid E2-a has the ability to destroy the cyst germinal cell and therefore inhibit the growth of cyst, and E2-a can reduce the number of PD-1+T cells, change the immune microenvironment and activate T cells. However, the molecular mechanism of inhibiting the growth of cysts and down-regulation of PD-1 by E2-a is not clear. Through E2-a intervention on vesicles of alveolar echinococcus and T cells respectively The objective of this study is to detect the receptor signaling pathway and regulating PD-1 transcription molecules of EmIR/EmER/EmTR involved in the proliferation of germinating cells, and to determine the effect of E2-a on the germinal cells and PD-1 target molecules. By using E2-a to intervene the alveolar echinococcosis model rats, the objective of vivo experiments is to verify the molecule mechanism of E2-a inhibiting the growth of germinal cells and PD-1 degradation, detect the local immune cells and molecules of granuloma of the cyst and therefore as well as clarify the effect of E2-a on the immune microenvironment, and therefore lays the theoretical foundation for the application of E2-a in the treatment of echinococcosis.

包虫病危害西部人群健康，制约经济发展，目前尚无疗效理想和安全的抗包虫病药物。研发既能对抗包囊生发细胞增殖、又能诱发宿主抗虫免疫的药物是控制包虫病策略之一。砂生槐种子是民间治疗虫毒的一味藏药，我们发现其种子水溶性生物碱E2-a能破坏包囊生发细胞、抑制包囊生长，同时E2-a能下调PD-1+T细胞数量，引起免疫微环境的改变，活化T细胞。然而E2-a抑制包囊生发细胞生长、下调PD-1的分子机制尚不清楚。本课题拟将泡球蚴囊泡、T细胞分别经E2-a干预，检测参与生发细胞增殖的EmIR/EmER/EmTR受体信号通路和调控PD-1转录的分子，确定E2-a作用于生发细胞、PD-1的靶分子；应用E2-a干预泡球蚴病模型鼠，体内试验验证E2-a抑制生发细胞生长、下调PD-1的分子机制，并检测包囊肉芽肿局部免疫细胞和分子，阐明E2-a对免疫微环境的调节作用，为E2-a在包虫病治疗中的应用奠定基础和和提供新的思路

包虫病危害西部人群健康，制约经济发展，目前尚无疗效理想和安全的抗包虫病药物。研发既能对抗包囊生发细胞增殖、又能诱发宿主抗虫免疫的药物是控制包虫病策略之一。砂生槐种子是民间治疗虫毒的一味藏药，我们发现其种子水溶性生物碱E2-a能破坏包囊生发细胞、抑制包囊生长，同时E2-a能下调调节性T细胞数量，引起免疫微环境的改变，活化T细胞。然而E2-a抑制包囊生发细胞生长的分子机制尚不清楚。本项目在体外将泡球蚴囊泡（生发细胞）经 E2-a 干预，检测 ERK 样蛋白激酶途径和 TGF-β样信号途径相关蛋白酶的水平；体外将 T 细胞经 E2-a 干预，检测其对T细胞生成及功能的影响；通过小鼠原头蚴感染模型，经 E2-a 干预后，验证其抑制囊泡生发细胞增殖、分化，并检测病变组织局部免疫细胞和分子。实验结果证明了砂生槐种子生物碱E2-a具有抗泡球蚴作用，而且可能是通过 EmIR/EmER/EmTR 受体相关信号通路，影响调节性T细胞生成而发挥作用；验证了，砂生槐种子生物碱E2-a 可以调控各类细胞因子，影响调节性T细胞的增殖、分化，进而通过对局部免疫微环境的调控完成抗泡球蚴作用，为E2-a在包虫病治疗中的应用奠定基础和和提供新的思路。