血必净对百草枯诱导人肾近曲小管上皮细胞-2氧化应激和炎症水平的影响

摘要

目的 观察血必净注射液对百草枯诱导的人肾近曲小管上皮细胞-2(HK-2)氧化应激和炎症水平的影响.方法 常规培养HK-2细胞,按随机数字表法分为空白对照组、百草枯中毒模型组、血必净预处理组、血必净对照组.采用高效液相色谱法(HPLC)检测细胞内百草枯含量;采用化学比色法检测超氧化物歧化酶(SOD)和丙二醛(MDA)水平;采用酶联免疫吸附试验(ELISA)检测白细胞介素-6(IL-6)和肿瘤坏死因子-α(TNF-α)水平.结果 随时间延长,百草枯中毒模型组和血必净预处理组HK-2细胞内百草枯含量逐渐升高,作用48 h达峰值,且血必净预处理组HK-2细胞内百草枯含量较百草枯中毒模型组明显降低(mg/L :4.26±0.20比5.77±0.18,P＜0.05).与空白对照组比较,百草枯中毒模型组MDA、TNF-α和IL-6含量明显升高〔MDA(nmol/mg):4.47±0.10比2.21±0.08,TNF-α(ng/L):206.91±13.22比98.14±5.67),IL-6(ng/L):253.33±5.22比116.97±13.54,均P＜0.05〕,SOD活性明显降低(U/mg :33.30±0.62比41.58±0.17,P＜0.05).与百草枯中毒模型组比较,血必净预处理组HK-2细胞内MDA、TNF-α和IL-6含量明显降低〔MDA(nmol/mg):2.92±0.17 比 4.47±0.10,TNF-α(ng/L):166.29±15.47 比 206.91±13.22,IL-6(ng/L):209.39±3.18 比253.33±5.22,均P＜0.05〕,SOD活性明显升高(U/mg :38.10±0.67比33.30±0.62,P＜0.05).结论 血必净可降低HK-2细胞内百草枯含量以及细胞氧化应激和炎症因子水平.%Objective To investigate the effect of Xuebijing on paraquat-induced oxidative stress and inflammation level in human kidney cell line-2 (HK-2) cells. Methods The HK-2 cells were routinely cultured and divided into blank control group, paraquat poisoning model group, Xuebijing injection pretreatment group and Xuebijing control group according to random number table method. The concentration of paraquat in HK-2 cells were measured by high performance liquid chromatography (HPLC); the activity of superoxide dismutase (SOD) and the level of malondialdehyde (MDA) were measured by chemical colorimetry method; the levels of interleukin-6 (IL-6) and tumor necrosis factor-α (TNF-α) were detected by enzyme linked immunosorbent assay (ELISA). Results With prolonged treatment, paraquat concentration gradually increased in HK-2 cells in the paraquat poisoning model group and the Xuebijing injection pretreatment group, reaching the peak value after treatment for 48 hours, moreover, the concentration of paraquat in HK-2 in the Xuebijing injection pretreatment group was significantly lower than that in the paraquat poisoning model group (mg/L: 4.26±0.20 vs. 5.77±0.18, P < 0.05). Compared with the blank control group, the contents of MDA, TNF-α and IL-6 of paraquat poisoning model group were obviously elevated [MDA (nmol/mg): 4.47±0.10 vs. 2.21±0.08, TNF-α (ng/L): 206.91±13.22 vs. 98.14±5.67, IL-6 (ng/L): 253.33±5.22 vs. 116.97±13.54, all P <0.05], and the activity of SOD was significantly reduced (U/mg: 33.30±0.62 vs. 41.58±0.17, P < 0.05). Compared with paraquat poisoning model group, the contents of MDA, TNF-α and IL-6 in Xuebijing injection pretreatment group were obviously decreased [MDA (nmol/mg): 2.92±0.17 vs. 4.47±0.10, TNF-α (ng/L): 166.29±15.47 vs. 206.91±13.22, IL-6 (ng/L): 209.39±3.18 vs. 253.33±5.22, all P < 0.05], and the activity of SOD was significantly increased (U/mg:38.10±0.67 vs. 33.30±0.62, P < 0.05) in the paraquat pretreatment group. Conclusion Xuebijing could reduce the concentration of paraquat in HK-2 cells, and decrease oxidative stress and inflammation factor levels in HK-2 cells.