Zhou Yuhua, Chen Chunyan, Li Qingtian, Sheng Huiqiu, Guo Xiaokui, Mao Enqiang
Department of Emergency, Ruijin Hospital, Shanghai Jiao Tong University School of Medicine, Shanghai, 200025, China.
Key Laboratory of Parasite and Vector Biology, Ministry of Health, Chinese Center for Tropical Disease Research, Shanghai Jiao Tong University School of Medicine, Shanghai, 200025, China.
Open Med (Wars). 2022 Apr 6;17(1):676-688. doi: 10.1515/med-2022-0446. eCollection 2022.
Acute lung injury (ALI) is a devastating human malignancy characterized by excessively uncontrolled inflammation and lung endothelial dysfunction. Non-coding RNAs play essential roles in endothelial protections during the pathological processes of ALI. The precise functions and molecular mechanisms of the lncRNA-NORAD-mediated endothelial protection remain obscure. This study reports NORAD was significantly induced in human pulmonary microvascular endothelial cells (HPMECs) under lipopolysaccharide (LPS) treatment. Silencing NORAD effectively protected HPMECs against the LPS-induced cell dysfunction. In addition, RNA pull-down and luciferase assay validated that NORAD sponged miR-30c-5p, which showed reverse functions of NORAD in the LPS-induced cell injury of HPMECs. Furthermore, the glucose metabolism of HPMECs was significantly elevated under LPS stimulation which promoted the glucose consumption and extracellular acidification rate (ECAR) of HPMECs. Inhibiting NORAD or overexpressing miR-30c-5p suppressed glucose metabolism in HPMECs, leading to protective effects on HPMECs under LPS stimulation. The glycolysis key enzyme, lactate dehydrogenase-A (LDHA), was subsequently identified as a direct target of miR-30c-5p. Finally, recovery of miR-30c-5p in NORAD-overexpressing HPMECs effectively overrode the NORAD-promoted glycolysis and impaired endothelial dysfunction under LPS stimulation by targeting LDHA. Summarily, we demonstrated a NORAD-miR-30c-5p-LDHA-glycolysis axis in the LPS-induced HPMECs dysfunction and , contributing to the development of anti-ALI therapeutic approaches.
急性肺损伤（ALI）是一种具有毁灭性的人类疾病，其特征为炎症过度失控和肺内皮功能障碍。非编码RNA在ALI病理过程中的内皮保护中发挥着重要作用。lncRNA-NORAD介导的内皮保护的确切功能和分子机制仍不清楚。本研究报道，在脂多糖（LPS）处理下，人肺微血管内皮细胞（HPMECs）中NORAD显著上调。沉默NORAD可有效保护HPMECs免受LPS诱导的细胞功能障碍。此外，RNA下拉和荧光素酶测定证实NORAD可吸附miR-30c-5p，在LPS诱导的HPMECs细胞损伤中，miR-30c-5p与NORAD发挥相反作用。此外，LPS刺激下HPMECs的葡萄糖代谢显著升高，促进了HPMECs的葡萄糖消耗和细胞外酸化率（ECAR）。抑制NORAD或过表达miR-30c-5p可抑制HPMECs的葡萄糖代谢，从而在LPS刺激下对HPMECs产生保护作用。糖酵解关键酶乳酸脱氢酶-A（LDHA）随后被确定为miR-30c-5p的直接靶点。最后，在过表达NORAD的HPMECs中恢复miR-30c-5p，通过靶向LDHA有效地抵消了NORAD促进的糖酵解，并改善了LPS刺激下的内皮功能障碍。总之，我们在LPS诱导的HPMECs功能障碍中证实了NORAD-miR-30c-5p-LDHA-糖酵解轴，这有助于抗ALI治疗方法的开发。
