mTOR-NLRP3炎症小体通路在急性高眼压视神经损伤中的作用及机制研究

Immunoinflammative injury is one of the vital mechanisms in glaucomatous retinal gangalion cells (RGCs) death, and mammalian target of rapamycin (mTOR) is a widely distributed serine/threonine protein kinase in cells. To date, relationship between immunoinflammation and mTOR is unclear. Recently, we have found that mTOR and NLRP3 were activated, played a crucial role during RGCs damage in mice acute ocular hypertension model. However, the mechanism needs further study. Therefore, we hypothesis that microglia activated, and promoted downstream inflammatory cytokines secretion via mTOR-NLRP3 inflammasome pathway in acute ocular hypertension model. To explore the profound mechanism, this study focus on “mTOR-NLRP3 inflammasome pathway” to clarify the molecular mechanism of this pathway based on mice acute ocular hypertension model and microglia oxygen-glucose deprivation (OGD) model by many kinds of technologies. Finally, from molecular, cellular and animal levels, we aim to explore the mechanism of how “mTOR-NLRP3 inflammasome pathway” mediates glaucomatous RGCs damage in acute ocular hypertension model, so far to reveal the RGCs injury mechanism of immunoinflammtion from a new prospective, and provide a new way of thinking for the prevention of glaucoma.

免疫炎症损伤是青光眼视网膜神经节细胞（RGCs）继发性损伤的关键机制之一，哺乳动物雷帕霉素靶蛋白（mTOR）是丝氨酸/苏氨酸蛋白激酶。但是mTOR与免疫炎症的关系仍未阐明。我们的前期发现急性高眼压模型中，mTOR和NLRP3炎症小体活化，在RGCs损伤过程中发挥重要作用。由此，我们提出假说：小鼠急性高眼压后，小胶质细胞活化，通过激活mTOR-NLRP3炎症小体通路，进而调节下游炎症因子的成熟和分泌，介导视网膜损伤和RGCs死亡。为论证此假说，本研究在小鼠急性高眼压模型和小胶质细胞OGD模型基础上，围绕“mTOR-NLRP3炎症小体通路”，采用多种检测技术，从分子、细胞和动物水平探讨mTOR-NLRP3炎症小体通路在急性高眼压视神经损伤中的作用和调控机制。本项目从mTOR-NLRP3炎症小体这个新视角出发，揭示急性高眼压视神经免疫炎症损伤的作用机制，为青光眼防治提供新的思路。

青光眼是威胁人类视觉健康的最主要的不可逆性致盲性眼病，免疫炎症机制在其病理生理过程中发挥重要作用，但具体机制需进一步探讨。本项目利用小鼠急性高眼压和BV2细胞缺糖缺氧／再灌注模型，于体内体外模拟急性高眼压导致视网膜损伤。结果发现，急性高眼压模型导致视网膜损伤，同时伴随小胶质细胞活化、mTOR磷酸化水平增加及NLRP3炎症小体激活。通过对BV2细胞缺糖缺氧／再灌注模型中TLR4的沉默干预（siRNA），发现mTOR-NLRP3炎症小体通路的激活可通过TLR4进行调控。为进一步探讨mTOR-NLRP3炎症小体通路的激活在急性高眼压中的作用和调控机制，我们分别予mTOR抑制和NLRP3炎症小体抑制干预，结果发现，抑制mTOR后，NLRP3炎症小体激活受阻，炎症因子IL-1β表达减少，视网膜损伤减轻。抑制NLRP3炎症小体激活后，炎症因子IL-1β表达减少，对视网膜损伤起到了保护作用。该研究证实，免疫炎症损伤是高眼压视网膜损伤的重要机制, mTOR-NLRP3炎症小体通路在急性高眼压视网膜损伤中发挥重要作用。