甲基莲心碱在氧化应激中的作用机制研究

Mechanism of neferine in antioxidant stress

摘要:

目的 探究甲基莲心碱（Nef）抗氧化应激作用及其机制。方法 体外培养大鼠近端肾小管上皮 细胞（NRK-52E），分为对照组、高渗组、高糖组、Nef 组、锌原卟啉Ⅸ（ZnP）组和HO-1 抑制剂组。采 用CCK-8 法检测细胞活力，Western blot 检测血红素加氧酶1（HO-1）和P67 蛋白的表达，DCFH-DA 探 针检测活性氧（ROS），化学氧化酶标法检测HO-1 酶活性，TBA 法检测丙二醛（MDA）。结果 与对照组 比较，高糖组的氧化应激指标P67 表达增加，ROS 和MDA 含量升高，细胞活力下降（P <0.05）；与高糖组比 较，Nef 组HO-1 蛋白表达和酶活性增加，P67 表达降低，ROS 和MDA 含量减少，细胞活力上升（P <0.05）； HO-1 抑制剂组较Nef 组上述作用减弱（P <0.05）。结论 Nef 通过诱导HO-1 表达并增加其酶活性，从而 减轻高糖培养的NRK-52E 细胞氧化应激损伤。

Abstract:

Objective To investigate the effect of neferine (Nef) on oxidative stress and its mechanism. Methods NRK-52E cells (normal rat kidney proximal tubular epithelial cells) were cultured in vitro and divided into six groups: control group, Mannitol group, high-glucose group (HG), HG+Nef group, zinc protoporphyrin IX group (ZnP) and HG+Nef+ZnP group. Cell viability was detected by CCK-8. The expressions of heme oxygenase-1 (HO-1) and p67 were detected by Western blot. Reactive oxygen species (ROS) was detected by DCFH-DA probes. The level of malondialdehyde (MDA) was measured by TBA and the activity of HO-1 was measured by TBIL. Results Compared with the control group, the oxidative stress indexes p67, ROS and MDA were increased, but the cell viability was decreased in the HG group (P < 0.05). Compared with the HG group, the expression and activity of HO-1 and cell viability were increased, but the expression of p67 and the levels of ROS and MDA were decreased in the HG+Nef group (P < 0.05). The effect of neferine mentioned above was reduced in the HG+Nef+ZnP group, compared with the HG+Nef group (P < 0.05). Conclusions Neferine attenuates oxidative stress injury in NRK-52E cells by inducing HO-1 expression and increasing its enzymatic activity.