Objective To investigate the influence of eicosapentaenoic acid(EPA)and docosahexaenoic acid(DHA)on the growth of HT-29 cell by regulating macrophage phenotype.Methods Human monocyte THP-1 was induced into M2-type macrophage,and then co-cultured with human colon cancer cell HT-29.The co-culture system was treated with DHA or EPA 0,10,20 and 40 μmol/L for 48 hours,respectively.The proliferation of HT-29 cell was detected by MTT,and the levels of TNF-α,IL-6,IL-1β and IL-10 in the co-culture supernatant were detected by ELISA.Real-time fluorescence quantitative PCR was used to detect the mRNA expressions of CD86,inducible nitric oxide synthase(iNOS),arginase 1(Arg1)and CD206 in macrophage.The protein expressions of phosphorylated NF-κB p65(p-NF-κB p65),phosphorylated signal transduction and activator of transcription 3(p-STAT3)and NOD-like receptor pyrin domain containing 3(NLRP3)in HT-29 cell were detected by Western blot.Results Compared with DHA 0 μmol/L,DHA 10,20 and 40 μmol/L reduced the cell survival rate,the level of IL-10 in supernatant,and the mRNA expressions of Arg1 and CD206,and increased the levels of TNF-α,IL-6 and IL-1β in supernatant,the mRNA expressions of CD86 and iNOS,and the protein expressions of p-NF-κB p65,p-STAT3 and NLRP3 in a concentration-dependent manner(P＜0.05).Compared with EPA 0 μmol/L,EPA 10,20 and 40 μmol/L reduced the cell survival rate,the level of IL-10 in supernatant,and the mRNA expressions of Arg1 and CD206,and increased the levels of TNF-α,IL-6 and IL-1β in supernatant,the mRNA expressions of CD86 and iNOS,and the protein expressions of p-NF-κB p65,p-STAT3 and NLRP3 in a concentration-dependent manner(P＜0.05).Conclusion DHA and EPA inhibit the growth of HT-29 cell by promoting the transformation of M2-type macrophage to M1-type macrophage.
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