重组人B型利钠肽抑制氧化低密度脂蛋白诱导的巨噬细胞氧化应激及炎症反应

Inhibition effect of recombinant human brain natriuretic peptide in oxidised low-density-liporotein induced oxidative stress and inflammatory respones of macrophages

目的探讨重组人B型利钠肽(rh BNP)对氧化低密度脂蛋白(ox-LDL)诱导的巨噬细胞氧化应激和炎症反应的抑制作用。方法建立ox-LDL干预THP-1源巨噬细胞的模型,分为3组:对照组、100μg/ml ox-LDL组和100μg/ml ox-LDL+10-9mol/L rh BNP组。收集各组细胞,应用共聚焦显微镜测定细胞内活性氧(ROS)水平,采用逆转录聚合酶链反应法(RT-PCR)及生物化学法分别检测超氧化物歧化酶(SOD)和丙二醛(MDA)基因的表达及活性,采用酶联免疫吸附法检测白细胞介素37(IL-37)的水平。结果与对照组相比,100μg/ml ox-LDL组的ROS水平升高(527. 30%±36. 20%比100. 00%±0. 00%),SOD mRNA的表达和活性降低[0. 53±0. 18比1. 00±0. 00;(256. 60±8. 20) U/ml比(355. 80±9. 58) U/ml],MDA mRNA的表达和含量升高[1. 59±0. 23比1. 00±0. 00;(29. 40±1. 68) nmol/ml比(5. 94±0. 51) nmol/ml],IL-37水平降低[(48. 05±3. 01) ng/L比(57. 82±2. 26) ng/L](均为P <0. 05);与100μg/ml ox-LDL组相比,100μg/ml ox-LDL+10-9mol/L rh BNP组的上述指标均有所改善[237. 30%±30. 62%,0. 90±0. 07,(310. 40±2. 97) U/ml,1. 14±0. 10,(20. 54±1. 55) nmol/ml,(53. 06±1. 87) ng/L,均为P <0. 05]。结论 rh BNP可能通过提高SOD表达及活性,上调IL-37水平,而抑制ox-LDL诱导的THP-1源巨噬细胞的氧化应激及炎症反应。

Objective To investigate the inhibition effect of recombinant human brain natriuretic peptide (rhBNP) in regulating oxidised low density lipoprotein (ox-LDL) induced oxidative stress and inflammatory responses in macrophage. Methods A model of ox-LDL-induced macrophage injury was established to evaluate the effect of rhBNP and divided into 3 groups: control group, 100 μg/ml ox-LDL group, 100 μg/ml ox-LDL+10-9 mol/L rhBNP group. Confocal microscopy was used to determine the cellular reactive oxygen species (ROS) levels. Reverse transcription polymerase chain reaction (RT-PCR) and colourimetry were used to detect the mRNA expression and activity, respectively, of superoxide dismutase (SOD) and malonaldehyde (MDA). Additionally, use enzyme-linked immunosorbent method (ELISA) to measure the IL-37 levels in cell culture medium of each group. Results Compared with the control group, intracellular ROS levels increased (527.30%±36.20% vs. 100.00%±0.00%), SOD mRNA expression and activity decreased [0.53±0.18 vs. 1.00±0.00,(256.60±8.20)U/ml vs.(355.80±9.58)U/ml], MDA mRNA expression and content increased [1.59±0.23 vs. 1.00±0.00,(29.40±1.68)nmol/ml vs.(5.94±0.51)nmol/ml], the level of IL-37 decreased [(48.05±3.01)ng/L vs.(57.82±2.26)ng/L] in the 100 μg/ml ox-LDL group (all P<0.05); these effects were significantly counteracted by 10-9 mol/L rhBNP [237.30%±30.62%, 0.90±0.07,(310.40±2.97)U/ml, 1.14±0.10,(20.54±1.55)nmol/ml,(53.06±1.87)ng/L, all P<0.05]. Conclusions rhBNP may up-regulate IL-37 levels by increasing SOD expression and activity, and inhibit ox-LDL-induced macrophage oxidative stress and inflammatory.