Protection Mechanisms Of EPA And DNA On Rat Mesangial Cells Stimulated By LPS

BackgroundRenal diseases such as IgA nephropathy, glomerular nephritis is a common primary glomerulonephritis, some patients may develop into end stage renal disease which endangers heath. Factors such as transforming growth factor beta 1 (TGF-β1), integrin linked kinase (ILK),β1 integrin and peroxisome proliferator-activated receptor y (PPARy) have a very close relationship with renal diseases. Meanwhile, the accumulation of extracellular matrix (ECM) protein components caused by metabolic imbalance like increased synthesis or decreased degradation is the main pathological features of glomerulosclerosis.Fish oil is extracted from deep-sea fish, and is rich in n-3 polyunsaturated fatty acids (n-3 PUFA). Its main active ingredients are eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA). Besides the anti-thrombotic effect, vasodilation, blood lipoid adjustment and anti-tumor effect, fish oil has a renal protective effect. In this study, we investigated the effects of EPA and DHA on the expression of MMPs, PPARy, ILK,β1 integrin and TGF-β1 in glomerular mesangial Cells (GMCs) stimulated by LPS (LPS) and to explore its possible protection mechanisms.Methods The mesangial cells in logarithmic growth phase were used. After serum-free cultured for 24 h, the cells were synchronized in the Go period, and then divided into six groups:the control group; LPS group (model group, LPS 10.0 mg/L); LPS plus EPA (10μmol/L,100μmol/L) group; LPS plus DHA (10μmol/L,100μmol/L) group. All the cells were cultured in 37℃and 5% CO2 for 24 h or 48 h.1 The TGF-β1 secretion was measured by enzyme linked immunosorbent assay (ELISA);2 The MMP-2,9 concentration in supernatant was measured by gelatin zymography;3 The expression of PPARγand ILK was determined by Western Blotting;4 The mRNA expression of PPARγ, ILK,β1 integrin and TGF-β1 were determined by RT-PCR;Results1 Effect of EPA and DHA on the expression of TGF-β1 in GMCs stimulated by LPSLPS could promote TGF-β1 secretion and mRNA expression; EPA and DHA could inhibit the GMCs TGF-β1 secretion and mRNA expression stimulated by LPS.2 Effects of EPA and DHA on the Secretion of MMP-2,9 in GMCs stimulated by LPSLPS could inhibit MMP-2,9 secretion; EPA and DHA could promote the GMCs MMP-2,9 secretion stimulated by LPS.3 Effects of EPA and DHA on the PPARγexpression in GMCs stimulated by LPSLPS could inhibit PPARγprotein and mRNA expression; EPA and DHA could promote the GMCs PPARγprotein and mRNA expression stimulated by LPS.4 Effects of EPA and DHA on the ILK expression in GMCs stimulated by LPSLPS could promote ILK protein and mRNA expression; EPA and DHA could inhibit the GMCs ILK protein and mRNA expression stimulated by LPS.5 Effects of EPA and DHA on the mRNA expression ofβ1 integrin in GMCs stimulated by LPSLPS could promoteβ1 integrin mRNA expression; EPA and DHA could inhibit the GMCsβ1 integrin mRNA expression stimulated by LPS.Conclusion1 EPA and DHA could promote the GMCs MMP-2,9 secretion stimulated by LPS.2 EPA and DHA could inhibit the GMCs TGF-β1 secretion and mRNA expression stimulated by LPS.3 EPA and DHA could promote GMCs PPARy protein and mRNA expression stimulated by LPS.4 EPA and DHA could inhibit the GMCsβ1 integrin and ILK protein and mRNA expression stimulated by LPS.