Abstract: The aim of this study is to investigate the effect of long non-coding RNA PLUTO on β cell function and its underlying mechanism. Islet cells of db/db, ob/ob and HFD mice were isolated and extracted by enzymatic method to detect the expression of PLUTO in islet cells of different obesity model mice. Glucolipid toxicity was used to stimulate islet primary cells and Min6 cells, and the expression of PLUTO was detected to elucidate the reasons of PLUTO down-regulation induced by obesity factors. RT-qPCR and ELISA were performed to analyze the function of PLUTO on β cells; Western blot and rescue experiments were carried out to elucidate the regulatory mechanism of β cells by PLUTO. The results showed that the expression levels of PLUTO were significantly decreased in obesity model mice compared with control mice; over-expression PLUTO in primary islets and Min6 cells could improve insulin biosynthesis and secretion; Western blot analysis showed that PLUTO promoted insulin secretion and synthesis by up-regulating the transcription and translation of Pdx1, which is the adjacent gene of PLUTO.