lncBRM调控YAP1蛋白翻译后修饰在同型半胱氨酸引起内皮凋亡和平滑肌细胞增殖并存中的机制研究

Vascular endothelial cells (ECs) apoptosis and smooth muscle cells (VSMCs) proliferation are fundamental characteristics of homocysteine (Hcy)-induced atherosclerosis, but it has not been solved yet that why happened different pathological changes of these two cells under the same condition. lncRNA was involved in the regulation of disease by protein posttranslational modification and other ways, and YAP1 has a dual role in regulating proliferation and apoptosis, but whether and how LncRNA regulates post-translational modification of YAP1 protein to mediate Hcy-induced the coexistence of EC apoptosis and VSMCs proliferation remains unclear. Therefore, we first explored the role of YAP1 in the coexistence of EC apoptosis and VSMCs proliferation induced by Hcy. Next, screened and identified lncBRM as its specific lncRNA and YAP1 as its functional target gene. Subsequently, revealing the mechanism that CRM1 synergistic Huh mediated lncBRM nuclear transport; To clarify the potential difference and mechanism that Hcy via lncBRM regulating GSK-3β and PRMT1 expression to mediate YAP1 protein phosphorylation of serine residues 412 and arginine residues 58 and 444 loci methylation to contribute to the coexistence of EC apoptosis and VSMCs proliferation, explore the key target, which can provide potential drug targets for the prevention and treatment of As.

血管内皮细胞(EC)凋亡和平滑肌细胞(VSMCs)增殖并存是同型半胱氨酸(Hcy)引起动脉粥样硬化(As)的基本特征，但同种条件下为何两类细胞病理变化迥异至今无解。lncRNA可通过蛋白翻译后修饰等多种方式参与疾病调控，而YAP1是一种具有增殖和凋亡双向调控作用的功能蛋白，但lncRNA是否及如何调控YAP1蛋白翻译后修饰介导Hcy引起EC凋亡和SMC增殖并存尚未清楚。故本课题首先探讨YAP1在Hcy引起EC凋亡和SMC增殖并存中的作用，筛选和确定lncBRM是其特异性lncRNA且YAP1是其功能性靶基因；揭示CRM1协同Huh介导核内lncBRM转运出核的作用机理；阐明Hcy经lncBRM调控GSK-3β和PRMT1表达介导YAP1蛋白丝氨酸残基412位点磷酸化和精氨酸残基58、444位点甲基化修饰引起血管EC凋亡和SMC增殖并存的潜在差异及机制，探寻关键靶点，为防治As提供潜在药靶。

背景：血管内皮细胞(EC)凋亡和平滑肌细胞(VSMCs)增殖并存是同型半胱氨酸(Hcy)引起动脉粥样硬化(As)的基本特征，但同种条件下为何两类细胞病理变化迥异至今无解。lncRNA可通过蛋白翻译后修饰等多种方式参与疾病调控，而YAP1是一种具有增殖和凋亡双向调控作用的功能蛋白，但lncRNA是否及如何调控YAP1蛋白翻译后修饰介导Hcy引起EC凋亡和SMC增殖并存尚不清楚。.内容：本课题通过复制ApoE-/-鼠高Hcy血症As动物模型，RNA-Seq等技术筛选并确定Hcy引起内皮细胞(ECs)凋亡和平滑肌细胞(VSMCs)增殖中的特异性lncRNA；沉默lncBRM后检测内皮细胞(EC)凋亡和平滑肌细胞(VSMCs)增殖变化，明确lncRNA在Hcy引起检测内皮细胞(EC)凋亡和平滑肌细胞(VSMCs)增殖中的作用；生物信息学在线软件分析lncBRM的功能性靶蛋白(提示为YAP1)，生物信息学分析YAP1精氨酸可能甲基化位点，westenblot检测沉默lncBRM后YAP1R58、444、S412位点的改变。.结果：首先，RNA-Seq结果显示LncBRM是Hcy引起内皮细胞(ECs)凋亡和平滑肌细胞(VSMCs)增殖中的特异性lncRNA，沉默lncBRM后可以抑制ECs凋亡和VSMCs增殖；其次，流式细胞术和EdU染色结果表明沉默lncBRM可以抑制ECs凋亡和VSMCs增殖；最后，生物信息学分析发现，干扰lncBRM之后，YAP1R58、444、S412位点表达降低，给予Hcy之后，YAP1R58、444、S412位点的表达水平有所升高。.该项目已见刊发表SCI论文6篇，申请国家专利2个，培养博士研究生2人、硕士研究生3人。.意义：本课题的实施将有利于阐明Hcy引起As的分子机制，确定关键靶点，为As的靶向治疗提供新的干预途径。

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