| Objectives①To observe if adenovirus mediated IL-24 gene transfer can inhibit LPS-induced proliferation of glomeru lar mesangial cells and to find out the effect of IL-24 on cell cycle regulatory protein p21,p27 and CyclinE of GMCs induced by LPS;②To explore the effect of IL-24 on GMCs excretion cytokines like IL-6 and TNF-α.Methods Ad.IL-24 and Ad.GFP were constructed through RT-PCR and amplify- cated in 293 cells. GMCs were infected with Ad.IL-24 or Ad.GFP and cultured in DMEM supplemented with 5%FCS/DMEM in the presence or absence of 10mg/L of LPS respectively. The mRNA expression of MDA7/IL-24 in GMC cells was confirmed by RT-PCR and the protein expression was confirmed by ELISA. Proliferation of GMC was measured by MTT assay and flow cytometry. The expression of cell cycle regulatory protein like p21, p27 and cyclinE were detected by Western-blotting .Results No IL-24 mRNA and protein expression were observed in cultured GMCs and even in LPS-activated GMCs. However, stable mRNA expressions of exogenous IL-24 gene were found in GMCs cultured for 4h to 48h. The levels of IL-24 in the supernatant were 79±3.61pg/ml at 24h and 98±2pg/ml at 48h respectively through ELISA. Adenovirus mediated IL-24 gene transfer didn't induce abnormal proliferation of GMC, but can inhibit LPS-induced proliferation of glomerular mesangial cells. The expression of p21 and p27 were down-regulated while CyclinE expression was up-regulated in GMC by LPS. Adenovirus mediated IL-24 gene transfer could abolish the role of LPS on regulation of p21, p27 and CyclinE expression. Finally,IL-24 gene delivery into GMC significantly increased the secretion of interleukin-6 ,but showed no impact on TNF-αexcretion. Conclusion It was concluded that adenovirus mediated exogenous IL-24 gene transfer could effectively inhibit LPS-induced proliferation of glomerular mesangial cells, and thus might find its clinical significance in gene therapy of mesangial cell proliferation.
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