lncRNACARLo-5调控miR-218在吸烟所致胰岛细胞凋亡和肝胰岛素抵抗中作用及干预研究

It has been proved that cigarette smoking can induce the apoptosis of islet cells and hepatic insulin resistance via causing inflammation, however, its molecular mechanisms remain unclear. Long non-coding RNA (lncRNAs) and microRNAs (miRNAs) have many important biological functions, but there are very few investigative publications about their roles and mechanisms in the damages induced by environmental pollutants and the development of diabetes. Based on our preliminary results that cigarette smoking and cigarette smoke extract (CSE) could induce the disorder of insulin secretion and insulin resistance in the human body, islet, pancreatic INS-1 cells, and liver L-02 cells in population study and cell experiments, we are going to investigate the roles and Andrographolide interventions of lncRNA CARLo-5 -regulated miR-218 in the apoptosis of islet cells and hepatic insulin resistance induced by cigarette smoking on the levels of cell, animal, and population, respectively, by using the following three levels. (i) CSE-induced damages of pancreatic INS-1 cells and liver L-02 cells in the presence or absence of anti-inflammatory factor, the siRNA or high expression plasmid of lncRNA CARLo-5, miR-218 mimic or inhibitor, and Andrographolide, respectively; (ii) Cigarette smoke promoting rat diabetes induced by the diet of high glucose and fat and Streptozotocin; (iii) Cohort study on population with different cigarette smoking levels and diabetes. Our results will be helpful to understand the molecular mechanisms underlying hazard induced by cigarette smoking and its role in the development of diabetes.

吸烟可通过引起炎症而诱导胰岛细胞凋亡和肝胰岛素抵抗，但其分子机制尚不清楚。长链非编码RNA (lncRNAs)和微小RNA(miRNAs)在环境污染物所致损害和糖尿病发生发展中的作用及机制研究报道甚少。本项目在人群研究和细胞实验发现吸烟和香烟烟雾抽提物(CSE)可引起人体、胰岛、胰INS-1和肝L-02细胞产生胰岛素分泌障碍和胰岛素抵抗的基础上，应用抗炎因子、lncRNA CARLo-5 siRNA和高表达质粒、miR-218 mimic和抑制剂及穿心莲内酯等处理CSE所致胰INS-1和肝L-02细胞损害、香烟烟雾促进高糖高脂饲料+链脲菌素所致大鼠糖尿病及不同程度吸烟和糖尿病人群研究，从正向和反向在细胞、动物和人群水平探讨lncRNA CARLo-5调控miR-218在吸烟所致胰岛细胞凋亡和肝胰岛素抵抗中作用及穿心莲内酯的干预效果，以揭示吸烟所致危害及其在糖尿病发生发展中的部分分子机制。

吸烟可通过引起炎症而诱导胰岛细胞凋亡和肝胰岛素抵抗，但其分子机制尚不清楚。长链非编码RNA(lncRNAs)和微小RNA(miRNAs)在烟草烟雾所致损害和糖尿病发生发展中的作用及机制研究报道甚少。本项目在人群研究和细胞实验发现吸烟和香烟烟雾抽提物(CSE)可引起人体、胰岛、胰INS-1和肝L-02细胞产生胰岛素分泌障碍和胰岛素抵抗的基础上，探讨lncRNA MALAT1调控miR-17、miR-191在烟草烟雾引起的胰岛细胞凋亡和肝胰岛素抵抗中的作用及穿心莲内酯的干预效果。主要从三个层次开展研究工作：(i) 在细胞水平探讨应用抗炎因子、lncRNA MALAT1 siRNA和高表达质粒、miR-17 mimic、miR-191 mimic抑制剂及穿心莲内酯等处理CSE所致胰INS-1和肝L-02细胞损害；(ii)香烟烟雾促进高糖高脂饲料+链脲菌素所致小鼠糖尿病及穿心莲内脂的干预效果；(iii) 不同程度吸烟和糖尿病人群研究；从正向和反向在细胞、动物和人群水平探讨lncRNA MALAT1调控miR-17、miR-191在吸烟所致胰岛细胞凋亡和肝胰岛素抵抗中作用及穿心莲内酯的干预效果。研究结果显示：(i)CSE可引起胰岛细胞凋亡和肝胰岛素抵抗，lncRNA MALAT1调控miR-17、miR-191可能是该作用的重要分子机制；(ii)香烟烟雾可加重糖尿病小鼠的葡萄糖代谢紊乱，穿心莲内酯可拮抗这种作用；(iii) 验证了吸烟与人群血糖水平升高、胰岛素水平降低及糖尿病患者有关，且lncRNA MALAT1调控 miR-17可能是其重要的分子机制。该研究揭示吸烟所致危害及其在糖尿病发生发展中的部分分子机制，为寻找吸烟所致糖尿病的早期生物标志及发现新的防控措施提供科学依据。