溶血磷脂酸对人单核细胞株基质金属蛋白酶-9和Toll样受体4表达的影响以及核因子-κB抑制剂的干预作用

Influence of lysophosphatidie acid on experssion of matrix metalloproteinases-9 and toll like receptor-4 in THP-1 cell and intervention of nuclear factor-κB inhibitor

目的探讨溶血磷脂酸(LPA)对人单核细胞株(THP-1)基质金属蛋白酶(MMP)-9和Toll样受体4(TLR-4)表达的影响以及核因子(NF)-κB抑制剂的干预作用。方法分别以0、0.1、0.5、1、5、10μmol/L的LPA加入人THP-1细胞4 h,以及将NF-κB抑制剂咖啡酸苯乙酯(CAPE)20 mg/L预处理THP-1细胞1 h后,再加入LPA1μmol/L4 h。应用酶联免疫吸附法测定细胞上清液的MMP-9含量,RT-PCR法测定TLR-4mRNA表达,Western Blot检测NF-κB p65活性。结果 LPA0、0.1、0.5、1、5、10μmol/L组细胞上清液MMP-9水平分别为(256.63±20.51)ng/ml、(296.57±10.92)ng/ml、(330.73±9.05)ng/ml、(367.8±6.4)ng/ml、(316.4±4.87)ng/ml及(303.00±6.45)ng/ml,各组间差异有统计学意义(均P<0.01);各组TLR-4 mRNA表达的差异有统计学意义(均P<0.01)。CAPE干预组细胞上清液MMP-9含量、TLR-4 mRNA表达及NF-κBp65活性显著低于LPA1μmol/L组(均P<0.01)。结论 LPA能促进THP-1细胞MMP-9和TLR-4的表达,NF-κB抑制剂预处理可以降低其表达水平。

Objective To explore the influence of lysophosphatidie acid（LPA） on experssion of matrix metalloproteinases（MMP）-9 and toll like receptor（TLR）-4 in THP-1 cell and intervention of nuclear factor（NF）-κB inhibitor.Methods The different contents of LPA（0,0.1,0.5,1,5,10 μmol/L） were added respectively into human THP-1 cells for 4 h;and the other THP-1 cells were pretreatment with CAPE（20 mg/L） for 1 h and then were stimulated with LPA（1 μmol/L） for 4 h.In the cell supernatant,the content of TLR-4 mRNA was measured by RT-PCR;the expression of MMP-9 was detected by enzyme-linked immunosorbent assay;the activity of NF-κB p65 subunit was tested by Western Blot.Results The expression of MMP-9 in LPA groups（0,0.1,0.5,1,5,10 μmol/L） were（256.63±20.51）ng/ml,（296.57±10.92）ng/ml,（330.73±9.05）ng/ml,（367.8±6.4）ng/ml,（316.4±4.87）ng/ml and（303.00±6.45）ng/ml,respectively.The expression of MMP-9 and TLR-4 mRNA were statistically differences between each group（all P0.01）.The expression of MMP-9,TLR-4 mRNA and the activity of NF-κB p65 in CAPE intervention group were significantly lower than those in LPA 1 μmol/L group（all P0.01）.Conclusion LPA can increase the expression of MMP-9,TLR-4 mRNA in THP-1 cells;and those can be reduced by the pretreatment of NF-κB inhibitor.