红景天苷对内毒素诱导的RAW264.7细胞损伤拮抗作用的研究

目的:观察不同浓度的红景天苷（SDS）对内毒素（LPS）引起的巨噬细胞RAW264.7损伤的拮抗作用，并初步探讨其作用机制。方法: 将小鼠巨噬细胞RAW264.7随机分为正常对照组（A组）、LPS组(B组)、SDS (5 μg/ml)对照组（C组）、SDS(5 μg/ml)+LPS组（D组）、SDS(10 μg/ml) 对照组（E组）、SDS(10 μg/ml)+LPS组(F组)、SDS(20 μg/ml) 对照组(G组)和SDS(20 μg/ml)+LPS组(H组)。选取对数生长期的细胞，相继用SDS（0~20 μg/ml）预处理1 h及1 μg/ml LPS刺激24 h后，收集细胞，用MTT比色法检测细胞的活力。6 h后收集细胞用Western blot检测细胞中NF-κB的含量，用ELISA法检测细胞上清中TNF-α、IL-6和IL-10的含量，同时用光密度法检测细胞上清中LDH的含量。结果: LPS可显著降低细胞的活力（P<0.05,P<0.01），增加NF-κB的表达（P<0.01），并显著增加细胞上清中TNF-α、IL-6、IL-10和LDH的含量（P<0.05,P<0.01）。而SDS则可以浓度依赖的方式对抗LPS所致细胞活力的减低、NF-κB含量的增加，使TNF-α、IL-6和LDH的含量明显减少，IL-10的含量明显增加；但不同浓度的SDS对正常细胞的上述指标没有影响。结论: SDS对LPS引起的细胞损伤具有对抗作用，其作用可能与其抑制炎症介质的释放有关。

Protective effects of salidroside on lipopolysaccharide-induced injury in RAW264.7 cells

AIM:To explore the protective effects and the underlying mechanism of salidroside (SDS) on lipopolysaccharide (LPS)-induced inflammation in murine macrophage RAW264.7 cells. METHODS: Cells were divided randomly into eight groups: control group (A group), LPS group (B group), SDS 5 group (C group), SDS 5+LPS group (D group), SDS 10 group (E group), SDS 10+LPS group (F group), SDS 20 group (G group) and SDS 20+LPS group (H group). After cells were stimulated with LPS (1 μg/ml) for 6 h in the presence or absence of 0-20 μg/ml SDS for 1 h, activation of nuclear factor kappa B (NF-κB) was detected and analyzed by Western blotting. Supernatant was then used for determination of TNF-α, IL-6, and IL-10 levels by ELISA and lactate dehydrogenase (LDH) activity with corresponding detection kit according to the manufacturer’s instructions. Cells were stimulated with LPS (1 μg/ml) for 24 h in the presence or absence of 0-20 μg/ml SDS for 1 h, and cell viability was measured using the MTT assay. RESULTS: MTT assay showed that 1 μg/ml LPS reduced cell viability, but SDS concentration-dependently reversed LPS-induced reduction of cell viability. Western blotting showed that SDS markedly inhibited the activation of NF-κB in LPS-treated cells. SDS concentration-dependently reduced LPS-induced TNF-α, IL-6, and LDH content and increased IL-10 content. CONCLUSION: Salidroside plays an important role in anti-inflammation of murine macrophage RAW264.7 cells induced by LPS. Protection may be correlated with inhibition of inflammatory factors through NF-κB signal pathway and the balance between pro- and anti-inflammatory systems.