Effect Of Glucose On Interleukin-18Gene Experssion Of Mouse Peritoneal Macrophages And Its Possible Mechanism

ObjectiveTo investigate the effect of high glucose(GLU) and fluctuant GLUconcentration on the expression of interleukin-18(IL-18) in mouse peritonealmacrophages(MPM); Observe the impact of time on the expression of IL-18and the correlation of JNK signal pathway on the expression of IL-18in highGLU treated MPM.Methods1Pairs of Kunming male MPM were isolated and Purified and were set up as theblank control group,4.0mmol/L GLU treatment group,8.0mmol/L GLUtreatment group,16.0mmol/L GLU treatment group,24.0mmol/L GLUtreatment group,32mmol/L GLU treatment group, and GLU variabilitytreatment group. The expression of IL-18mRNA and the secretion of IL-18were respectively measured by reverse-transcription-polymer chainreaction(RT-PCR) and enzyme linked immunosorbent assay(ELISA).1MPM which were obtained from Kunming male mouse,were randomly assigned tofour GLU bellow(n=3in each):3h group,6h group,12h group,18h group.The expression of IL-18mRNA and the secretion of IL-18were respectivelymeasured by RT-PCR and ELISA.2MPM which were obtained from Kunming male mouse, according to theconcentration of c-Jun N-terminal kinase(JNK) signaling inhibitor SP600125,were randomly assigned to four groups bellow(n=3in each): the blank controlgroup,5μ mol/L group,10μ mol/L group, and15μ mol/L group. Theexpression of IL-18mRNA were determined by RT-PCR and the secretion ofIL-18in supernatant were measured by immunological ELISA. Results1GLU increases IL-18mRNA expression dose-dependently (4,8,16,24and32mmol/l). The GLU (4mmol/l) induced a little IL-18mRNA expression,IL-18/-actin is0.13±0.04. Then accompany with the increase of GLUconcentration the IL-18mRNA expression rise correspondly. Fluctuant GLUconcentration induced a6.0-fold increase in IL-18gene expression, compared withthat under4mmol/L conditions (P <0.05).The ELISA results of IL-18wereconsistent with the results of RT-PCR, and the comparison of IL-18expressionbetween any high GLU groups is statistical significance (P <0.05).2From3h to12h, High glucose(HG) increase IL-18mRNA expression andsecretion in MPM Time-dependently-induced mRNA expression of IL-18genes,which occurred as early as3h, and some reached a peak at12h. TheHG-induced increase in mRNA expression declined at18h compared with6h（.P<0.001).The ELISA results of IL-18consistent with the results of RT-PCR,andthe comparison between any HG groups is statistical significance (P <0.01).3JNK inhibitor SP600125inhibited HG-induced IL-18gene expressiondose-dependently (5,10and15mmol/l) compared with the blank control group（P<0.05).The ELISA results of IL-18consistent with the results of RT-PCR,andthe comparison between any HG groups is statistical significance (P <0.05).Conclusions1HG increases IL-18expression dose-dependently. Fluctuant GLU concentrationplay a important role in HG-induced IL-18gene expression.2From3h to12h, the HG increases IL-18mRNA expression time-dependently.The HG increases IL-18mRNA expression declined at18h.3JNK signaling maybe implicated in the cascade of HG-induced IL-18geneexpression.