LncRNA AC068039.4通过miR-26a-5p调控TRPC6参与肺动脉高压肺血管重塑的机制

瞬时受体电位阳离子通道6（TRPC6）介导的外钙内流在低氧性肺动脉高压（HPH）肺血管重塑的发生中起重要作用，前期研究证实低氧上调TRPC6，但调控机制不明。预实验提示低氧上调LncRNA AC068039.4、下调miR-26a-5p，TRPC6是miR-26a-5p的靶基因。本项目拟在人肺动脉平滑肌细胞（PASMC）上建立AC068039.4沉默和过表达模型，探讨AC068039.4对细胞增殖和凋亡的影响；通过双荧光素酶报告基因等方法验证AC068039.4通过ceRNA模式调控TRPC6表达；建立小鼠基因敲除HPH模型，探究AC068039.4在肺血管重塑中的作用。从而在分子、细胞和动物水平论证“低氧上调LncRNA AC068039.4，通过ceRNA模式竞争性结合miR-26a-5p，上调TRPC6，促进PASMC增殖，抑制细胞凋亡，导致肺血管重塑”的假说，为HPH防治提供新思路

Extracellular calcium influx mediated via transient receptor potential cation channel 6 (TRPC6) plays an important role in the development of pulmonary vascular remodeling in hypoxic pulmonary hypertension (HPH). Previous studies have confirmed that hypoxia up-regulates TRPC6, but the regulatory mechanism is unknown. Pre-experiment suggests that hypoxia up-regulates LncRNA AC068039.4 and down-regulates miR-26a-5p, while TRPC6 is a target gene of miR-26a-5p. We will establish LncRNA AC068039.4 silencing and overexpression model on human pulmonary artery smooth muscle cells (PASMC) to investigate the effect of LncRNA AC068039.4 on cell proliferation and apoptosis. Then, we will use double luciferase reporter gene assay to verify that LncRNA AC068039.4 regulates TRPC6 expression through ceRNA mode. Next, we will explore the role of LncRNA AC068039.4 on pulmonary vascular remodeling via knockout mouse of HPH model. Our study will demonstrate the hypothesis that "hypoxia up-regulates LncRNA AC068039.4, and competitively binds to miR-26a-5p by ceRNA mode, up-regulates TRPC6, promotes PASMC proliferation and inhibits apoptosis, and leads to pulmonary vascular remodeling" from the aspects of the molecular, cellular and animal level, so as to provide new insights for the prevention and treatment of HPH.