lncRNA NEAT1通过靶向抑制miR-29a促进低氧低糖诱导的肺纤维化进程的机制研究

目的 探究lncRNA NEAT1通过负调控miR-29a促进低氧低糖诱导的肺纤维化进程的机制.方法 收集健康者和缺血再灌注引起急性肺损伤患者的血清样本,通过qRT-PCR检测血清样本中NEAT1与miR-29a的表达.构建肺纤维化细胞模型和动物模型,进行离体和在体的RNA干扰,通过显微镜观察细胞的形态学改变,Hoec...

Study on mechanism of lncRNA NEAT1 in promoting hypoxia and hypoglycemia induced pulmonary fibrosis by targetedly regulating miR-29a

Objective To investigate the mechanism by which lncRNA NEAT1 negatively regulates miR-29 a to promote the progression of pulmonary fibrosis induced by hypoxia and low glucose.Methods Serum samples from healthy subjects and patients with acute lung injury induced by ischemia-reperfusion were collected and the expression of NEAT1 and miR-29 a in serum samples was detected by qRT-PCR.The cell model and the animal model of pulmonary fibrosis cells were built;the in vitro and in vivo RNA was interfered;the morphologic change of cells was observed through microscope;the apoptosis was detected by Hoechst staining;the pathologic change of lung tissue was evaluated by HE staining and Masson staining;the degree of lung tissue fibrosis was determined by lung tissue hydroxyproline(HYP)content;the expression of fibrosis marker proteins in cells,such as col1,col3 a1,α-SMA and TGF-β,was detected by Western blot;the expression of NEAT1 and miR-29 a was detected by qRT-PCR;the interaction and targeting relationship between NEAT1 and miR-29 a were verified by RIP and dual luciferase reporter genes.Results In serum samples from patients with acute lung injury induced by ischemia-reperfusion,NEAT1 expression was up-regulated and mir-29 a expression was down-regulated.Up-regulation of NEAT1 and down-regulation of miR-29 a were also found in cell and animal models.In the cell model of pulmonary fibrosis induced by hypoxia and hypoglycemia,it was found that NEAT1 knockout could inhibit the change of alveolar epithelial cell morphology caused by hypoxia and hypoglycemia from cobblestone type to long-spindle type,inhibit the apoptosis of alveolar epithelial cells,and reduce the expression of fibrotic marker proteins col1,col3 a1,α-SMA and TGF-β.RIP experiment revealed the direct binding of NEAT1 to miR-29 a.Dual luciferase reporter gene detection revealed that NEAT1 targetedly regulated miR-29 a.Inhibition of miR-29 a could reverse the inhibitory effect of NEAT1 knockout on hypoxia and hypoglycemia-induced apoptosis of alveolar epithelial cells and reducing the expression of fibrotic marker proteins col1,col3 a1,α-SMA and TGF-β,which then promoted the progression of pulmonary fibrosis.Conclusion lncRNA NEAT1 promotes hypoxia-induced pulmonary fibrosis by targeted inhibition of miR-29 a,providing new ideas for studying the mechanism of hypoxia-induced pulmonary fibrosis and developing new drugs.