Protective Effects Of EPA And DHA On Rat Glomerular Mesangial Cells Stimulated By Lipopolysaccharide

ObjectiveTo investigate the effects of eicosapentaenoic acid(EPA) and docosahexaenoic acid(DHA) on matrix metalloproteinases(MMPs),tissue inhibitors of metalloprotein-ases(TIMPs),peroxisome proliferator-activated receptor gamma(PPARy),monocyte chemoattractant protein-1(MCP-1),transforming growth factor beta 1(TGF-β1) on rat glomerular mesangial cells(GMCs) stimulated by lipopolysaccharide(LPS), and explore the possible protective mechanism of EPA and DHA to impaired GMCs.Methods and Results1. Effect of EPA and DHA on the expression of MMP-2/TIMP-2 mRNA on GMCs stimulated by LPS by real-time PCRGMCs were cultured in vitro. Count the 4th generation cell at a 1×105per well to the kind of 6-well plates, and then divided into six groups:control group; LPS group(with LPS final concentration of 10μg-mL-1); EPA group(with EPA final concentration of 10μmol·L-1 and 100μmol·L-1); DHA group(with DHA final concentration of 10μmol·L-1 and 100μmol·L-1). The expression of MMP-2 and TIMP-2 mRNA of GMCs was measured by real-time PCR after incubated 48h.The result showed that the level of MMP-2 mRNA and the ratio of MMP-2/TIMP-2 were significantly attenuated, TIMP-2 mRNA significantly increased when stimulated by LPS(P＜0.01). EPA and DHA could decrease the level of MMP-2 and TIMP-2 mRNA, increase the ratio of MMP-2/TIMP-2. 2. Effect of EPA and DHA on the expression of MMP-9/TIMP-1 mRNA on GMCs stimulated by LPS by real-time PCRCount the 4th generation cell at a 1×105per well to the kind of 6-well plates, and then divided into the same six groups. The expression of MMP-9 and TIMP-1 mRNA of GMCs was measured by real-time PCR after incubated 48h.The result showed that the level of MMP-9 mRNA and the ratio of MMP-9/TIMP-1 were significantly attenuated, TIMP-1 mRNA significantly increased when stimulated by LPS(P＜0.01). EPA and DHA could decrease the level of MMP-9 and TIMP-1 mRNA, increase the ratio of MMP-9/TIMP-1.3. Effect of EPA and DHA on the expression of PPARy mRNA on GMCs stimulated by LPS by real-time PCRGMCs were cultured for 24h,48h,72h, respectively. The expression of PPARy mRNA of GMCs was measured by real-time PCR. The result showed that the level of PPARγmRNA was significantly attenuated when stimulated by LPS(P<0.01). EPA and DHA could increase the level of PPARy mRNA.4. Effect of EPA and DHA on the expression of MCP-1,TGF-β1 mRNA on GMCs stimulated by LPS by real-time PCRGMCs were cultured for 24h,48h,72h, respectively. The expression of MCP-1,TGF-β1 mRNA of GMCs was measured by real-time PCR. The result showed that the level of MCP-1,TGF-β1 mRNA was significantly increased when stimulated by LPS(P＜0.01). EPA and DHA could decrease the level of MCP-1,TGF-β1 mRNA.5. Effect of EPA and DHA on the level of PPARγ,TGF-β1 protein on GMCs stimulated by LPS by immunocytochemistryCount the cell at a 1×104per well to the kind of 24-well plates, and then divided into the same six groups. The expression of PPARγ,TGF-β1 protein of GMCs was detected by immunocytochemistry after incubated 48h. The result showed that the level of PPARy protein was significantly attenuated and TGF-β1 protein significantly increased when stimulated by LPS(P<0.05). EPA and DHA could increase the level of PPARy mRNA and decrease the level of TGF-β1 mRNA. Conclusion1. EPA and DHA can decrease the level of MMP-2,MMP-9,TIMP-1,TIMP-2 mRNA of GMCs stimulated by LPS, increase the ratio of MMP-2/TIMP-2,MMP-9/TIMP-1.2. EPA and DHA can increase the level of PPARy mRNA and protein of GMCs stimulated by LPS.3. EPA and DHA can decrease the level of MCP-1 mRNA of GMCs stimulated by LPS.4. EPA and DHA can decrease the level of TGF-β1 mRNA and protein of GMCs stimulated by LPS.