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目的探讨1型多聚ADP核糖聚合酶[poly(ADP-ribose)polymerase-1,PARP-1]对炎性刺激物脂多糖(lipopolysaccharide,LPS)诱导人外周血单个核细胞产生炎性细胞因子的作用及其机制。方法①对原代培养的正常人外周血单个核细胞给予LPS刺激24h后,采用酶联免疫吸附法(ELISA法)检测炎性细胞因子白细胞介素(IL)-1β、IL-6、IL-10、肿瘤坏死因子-α(TNF-α)的表达水平,结果于450nm波长处测定的光密度值(A值)表示,并观察PARP-1抑制剂3-氨基苯甲酰胺(3-aminobenzamide,3-AB)对上述炎性细胞因子表达的影响;②提取培养细胞全蛋白,采用Westernblotting法检测培养细胞内PARP-1蛋白表达水平及3-AB对PARP-1蛋白表达的影响,结果使用灰度值表示。结果空白对照组炎性细胞因子IL-1β、IL-6、IL-10、TNF-α表达水平(A值)分别为12.40±4.09、43.58±10.43、11.20±2.30、51.36±3.43;LPS刺激组分别为233.87±30.60、320.27±10.13、26.6±2.09、506.47±27.83,均显著高于空白对照组(P<0.01);3-AB抑制组分别为125.04±28.32、208.91±28.76、17.03±2.03、152.86±16.86,均显著低于LPS刺激组(P<0.01),但高于空白对照组。LPS刺激组PARP-1蛋白表达显著高于空白对照组(P<0.01);3-AB抑制组PARP-1蛋白表达显著低于LPS刺激组(P<0.01)。PARP-1的蛋白表达水平与炎性细胞因子IL-1β、IL-6、IL-10、TNF-α表达水平均具有明显相关性,r值分别为-0.619(P<0.05)、-0.750(P<0.05)、-0.593(P<0.05)、-0.694(P<0.05)。结论在LPS诱导人外周血单个核细胞炎性反应产生细胞因子过程中,PARP-1的表达水平与炎性细胞因子的表达水平显著相关。PARP-1可能是炎性反应免疫激活过程中的重要调节因子之一。
Objective To investigate the effect of poly (ADP-ribose) polymerase-1 (PARP-1) on proliferation of human peripheral blood mononuclear cells induced by lipopolysaccharide (LPS) Role and mechanism. Methods ① The primary cultured human peripheral blood mononuclear cells were stimulated with LPS for 24 hours. The levels of IL-1β, IL-6, IL-6 and IL-6 in inflammatory cells were detected by enzyme linked immunosorbent assay (ELISA) 10, the expression of tumor necrosis factor-α (TNF-α), the results of optical density measured at 450nm value (A value), and observed PARP-1 inhibitor 3-aminobenzamide (3-aminobenzamide 3-AB) on the expression of inflammatory cytokines; ②The total protein of cultured cells was extracted, the expression of PARP-1 in cultured cells and the effect of 3-AB on the expression of PARP-1 protein were detected by Western blotting. Degree value. Results The expression of IL-1β, IL-6, IL-10 and TNF-α in the blank control group were 12.40 ± 4.09, 43.58 ± 10.43, 11.20 ± 2.30 and 51.36 ± 3.43, respectively; (233.87 ± 30.60,320.27 ± 10.13,26.6 ± 2.09,506.47 ± 27.83, respectively) were significantly higher than those in the blank control group (P <0.01); the levels of 3-AB inhibition group were 125.04 ± 28.32,208.91 ± 28.76,17.03 ± 2.03, 152.86 ± 16.86, respectively, which were significantly lower than that of LPS stimulation group (P <0.01), but higher than that of blank control group. The protein expression of PARP-1 in LPS stimulation group was significantly higher than that in control group (P <0.01). The protein expression of PARP-1 in 3-AB inhibition group was significantly lower than that in LPS stimulation group (P <0.01). The protein expression of PARP-1 was significantly correlated with the expression of IL-1β, IL-6, IL-10 and TNF-αin the inflammatory cells. The r values were -0.619 (P <0.05) P <0.05), -0.593 (P <0.05), -0.694 (P <0.05). Conclusions The expression of PARP-1 is significantly associated with the expression of inflammatory cytokines in LPS-induced cytokine production by human peripheral blood mononuclear cells. PARP-1 may be one of the important regulators of immune activation during inflammatory response.