LncRNA-PVT1作为ceRNA调控mTOR信号通路参与哮喘气道平滑肌细胞增殖、凋亡与自噬的机制研究

LncRNA can be an endogenous competitive RNA，binding miRNA and regulating its downstream target genes，then being involved in the onset of disease process.Studies have confirmed that LncRNA-PVT1 expression is decreased in the asthmatic airway smooth muscle tissues, so what about its effect on the proliferation ,apoptosis and autophagy of airway smooth muscle cell? it have be known that LncRNA-PVT1 can be interact with mir - 1207-5p, At the same time , mir-1207-5p can regulate cell proliferation, apoptosis and autophagy by mTOR pathways，so could PVT1 affect mTOR pathway through mir - 1207-5p to regulate airway smooth muscle proliferation, apoptosis and autophagy,then being involved in airway remodeling?In this study, we will treat ASMCs with TGFβ after PVT1 gene interference and intervent the chronic OVA asthmatic rats model with Ad-PVT1.Then proliferation ,autophagy and apoptosis will be detect by transmission electron microscopy，Laser Capture microdissection and molecular biologytechnology .we will explore the possible mechanism of asthma airway remodeling’s onset from the angle of the epigenetic, provide the experimental basis for the LncRNA - PVT1 participate in airway remodeling and new targets for diagnosis and treatment of asthma.

LncRNA可以作为内源性竞争性RNA（ceRNA）结合miRNA调控其下游靶基因参与疾病的发病过程。已有研究证实在哮喘病人气道平滑肌细胞中LncRNA-PVT1表达减少,那么其对气道平滑肌细胞的增殖凋亡以及自噬有何影响?已知PVT1与mir-1207-5p存在相互作用位点，同时mir-1207-5p可以通过mTOR通路调节细胞的增殖，凋亡与自噬，那么PVT1是否可以通过mir-1207-5p影响mTOR通路调控的气道平滑肌细胞增殖、凋亡与自噬，从而参与气道重塑? 本项目拟通过TGFβ作用于PVT1基因干扰后的气道平滑肌细胞，及腺病毒包埋的siPVT1干预慢性OVA哮喘大鼠模型，应用分子生物学实验、激光捕获显微切割及透射电镜等技术检测细胞增殖凋亡与自噬，从表观遗传的角度探讨哮喘气道重塑发病可能机制，进一步为LncRNA-PVT1参与气道重塑提供实验依据，为哮喘诊治提供新的靶点。

LncRNA可以作为内源性竞争性RNA（ceRNA）结合miRNA调控其下游靶基因参与疾病的发病过程。已有研究证实在哮喘病人气道平滑肌细胞中LncRNA-PVT1表达减少,那么其对气道平滑肌细胞的增殖凋亡以及自噬有何影响?已知PVT1与mir-1207-5p存在相互作用位点，同时mir-1207-5p可以通过mTOR通路调节细胞的增殖，凋亡与自噬，那么PVT1是否可以通过mir-1207-5p影响mTOR通路调控的气道平滑肌细胞增殖、凋亡与自噬，从而参与气道重塑? 本研究通过TGFβ作用于PVT1基因干扰后的气道平滑肌细胞，及腺病毒包埋的siPVT1干预慢性OVA哮喘大鼠模型，应用分子生物学实验、激光捕获显微切割及透射电镜等技术检测细胞增殖凋亡与自噬，初步证实了siPVT1 可以降低增殖的气道平滑肌细胞数量、降低气道平滑肌细胞增殖相关基因及蛋白的表达，同时促进气道平滑肌细胞自噬。同时动物实验表明， Ad-shPVT1干预后可以抑制哮喘小鼠模型气道重塑、减轻气道炎症、促进气道平滑肌自噬及凋亡。本研究从表观遗传的角度探讨哮喘气道重塑发病的机制，为LncRNA-PVT1参与气道重塑提供实验依据，为哮喘诊治提供新的靶点。