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目的:观察白藜芦醇对佛波酯诱导的U937细胞中基质金属蛋白酶-9活性的影响,并从蛋白、mRNA及核转录因子激活蛋白-1(AP-1)水平对其影响进行分析。方法:酶谱法测定U937细胞培养上清中MMP-9的活性;Western blot法考察MMP-9蛋白的生成;RT-PCR法检测MMP-9 mRNA的表达;电泳迁移率变动分析法(EMSA)研究核转录因子SP-1的活性。结果:PMA 10nmol/L 可显著诱导无血清培养的U937细胞中MMP-9活性(P<0.01);白藜芦醇在1和10 μmol/L浓度下可抑制 PMA 10 nmol/L诱导的MMP-9活性(P<0.05 和P<0.01);PMA 10 nmol/L可显著诱导U937细胞中MMP-9蛋白生成(P<0.01)和MMP-9 mRNA的表达(P<0.01),白藜芦醇在1、10μmol/L浓度下可抑制PMA 10 nmol/L诱导的MMP-9蛋白生成和MMP-9 mRNA的表达(P<0.05);白藜芦醇在10、1和0.1μmol/L浓度下可抑制PMA诱导的U937细胞中AP-1的活化。结论:白藜芦醇可有效地抑制PMA诱导的U937细胞中MMP-9的活性,其作用可能是通过抑制PMA诱导的U937细胞核转录因子AP-1活化,进而降低MMP-9 mRNA表达,减少MMP-9蛋白生成而实现的。
OBJECTIVE: To observe the effect of resveratrol on the activity of matrix metalloproteinase-9 in U937 cells induced by phorbol ester, and to analyze the effect of protein, mRNA and nuclear transcription factor-1 (AP-1) levels on it. Methods: The activity of MMP-9 in culture supernatant of U937 cells was determined by zymography; the production of MMP-9 protein was detected by Western blot; The expression of MMP-9 mRNA was detected by RT-PCR; Electrophoretic mobility shift analysis (EMSA) The activity of nuclear transcription factor SP-1 was studied. RESULTS: PMA 10 nmol/L significantly induced MMP-9 activity in serum-free U937 cells (P <0.01); resveratrol inhibited PMA 10 nmol/L-induced MMP levels at 1 and 10 μmol/L. 9 activity (P<0.05 and P<0.01); PMA 10 nmol/L could significantly induce MMP-9 protein production (P<0.01) and MMP-9 mRNA expression in U937 cells (P<0.01), resveratrol Inhibition of PMA 10 nmol/L induced MMP-9 protein production and MMP-9 mRNA expression at concentrations of 1, 10 μmol/L (P<0.05); resveratrol at concentrations of 10, 1 and 0.1 μmol/L It can inhibit PMA-induced activation of AP-1 in U937 cells. CONCLUSION: Resveratrol can effectively inhibit the activity of MMP-9 in U937 cells induced by PMA. The effect of resveratrol may be through inhibiting PMA-induced activation of nuclear transcription factor AP-1 in U937 cells, thereby reducing the expression of MMP-9 mRNA and reducing MMP -9 protein production and achieved.