| Objective To investigate EPA and DHA on the proliferation,apoptosis and inflammatory response of rat glomerular mesangial cells(GMCs)stimulated by lipopolysaccharide(LPS)and explore the possible protect mechanism of EPA and DHA.Methods and Results1.The effect of EPA and DHA on the proliferation in normal GMCs.The proliferation of cultured glomerular mesangial cells was analysed by MTT in vitro:the 3rd generation cell was cultured in 96-well plates at 5×103 per well,and then divided into four groups:the control group;EPA(10-2mol·L-1,10-1mol·L-1)group; DHA(10-2mol·L-1,10-1mol·L-1)group.All the cells were cultured in 37℃for 24 h, 48h,72h.The proliferation was measured by MTT method.The results showed that there was no significant differences between the normal group and EPA,DHA groups.2.The effect of EPA and DHA on the proliferation in LPS-stimulated GMCs.The 3rd generation cell was used for experiments and was divided into four groups:the control group;LPS(10%FCS,with LPS(10μg/ml,final concentration); EPA(10-2mol·L-1,10-1mol·L-1);DHA(10-2mol·L-1,10-1mol·L-1)groups,all of them were cultured in 37℃for 24 h,48h,72h,The proliferation was measured by MTT method.The results showed that LPS could promote the proliferation of GMCs,EPA and DHA could inhibit the excessive proliferation in a dose-dependent manner(P＜0.01,P＜0.01).3.The effect of EPA and DHA on the apoptosis of LPS-stimulated GMCsAccording to apoptosis test kit,the apoptosis was detected by flow cytometry. LPS could inhibit the apoptosis of GMCs significantly.EPA and DHA(10-1mmol·L-1) could promote the apoptosis of GMCs.4.The effect of EPA and DHA on antioxidant system of mesangial cellsThe activity of SOD and GSH-PX,the level of MDA were measured according to the test kit.The results showed that at all time points,there was no significant difference of drug group SOD,MDA,GSH-PX with the normal group.5.The effect of EPA and DHA on the antioxidant system of GMCs induced by LPSThe activity of SOD and GSH-PX,the level of MDA were measured according to the test kit.The results showed that LPS could increase the activity of SOD and GSH-PX and increase the level of MDA in GMCs.EPA and DHA could decreased the activity of SOD and GSH-PX and decrease the level of MDA significantly compared with LPS Group(P＜0.01).6.The effect of EPA and DHA on the expression of MCP-1 and TGF-β1 in GMCs induced by LPSThe expression of MCP-1 and TGF-β1 was measured by immunohistochemistry method.The results showed that LPS could increase the expression of MCP-1 in GMCs.EPA and DHA could inhibit the expression significantly compared with LPS group.Conclusion1.EPA and DHA mesangial cells can inhibit the excessive proliferation and the promote the apoptosis of GMCs stimulated by LPS.2.EPA and DHA can increase the ability of antioxidition by inhance the activity of antioxidant enzymes(SOD,GSH-PX).3.EPA and DHA can reduce the expression of TGF-β1 MCP-1.
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