目的 研究龙胆苦苷对脂多糖（lipopolysaccharide，LPS）诱导血管内皮细胞损伤的作用及机制。方法 以LPS刺激大鼠主动脉血管内皮细胞RAEC，并给予低、中、高剂量（5、10、20 μmol/L）龙胆苦苷进行干预，采用CCK-8法检测细胞增殖情况；采用流式细胞术检测细胞凋亡情况；采用试剂盒检测细胞丙二醛水平和超氧化物歧化酶（superoxide dismutase，SOD）活性；采用qRT-PCR法检测细胞中肿瘤坏死因子-α（tumor necrosis factor-α，TNF-α）、白细胞介素-6（interleukin-6，IL-6）、IL-1β mRNA表达情况；采用Western blotting方法检测细胞中磷酸化p65（p-p65）和剪切型半胱氨酸蛋白酶-3（cleaved Caspase-3）蛋白表达情况。RAEC细胞给予核因子-κB（nuclear factor-κB，NF-κB）信号激活剂处理，考察NF-κB信号激活剂对龙胆苦苷改善RAEC细胞损伤的影响。结果 与对照组比较，模型组细胞活性显著降低（P<0.05），细胞凋亡率显著升高（P<0.05），丙二醛水平显著升高（P<0.05），SOD活性显著降低（P<0.05），TNF-α、IL-6和IL-1β mRNA表达水平显著升高（P<0.05），cleaved Caspase-3和p-p65蛋白表达水平显著升高（P<0.05）。与模型组比较，龙胆苦苷组细胞活性显著升高（P<0.05），细胞凋亡率显著降低（P<0.05），丙二醛水平显著降低（P<0.05），SOD活性显著升高（P<0.05），TNF-α、IL-6和IL-1β mRNA表达水平显著降低（P<0.05），cleaved Caspase-3和p-p65蛋白表达水平显著降低（P<0.05）。NF-κB信号激活剂显著抑制龙胆苦苷改善RAEC细胞损伤的作用（P<0.05）。结论 龙胆苦苷能够抑制LPS诱导的RAEC细胞凋亡、氧化损伤和炎性因子表达，其作用机制可能与抑制NF-κB信号通路有关。

Objective To study effect and mechanism of gentiopicroside on lipopolysaccharide (LPS)-induced vascular endothelial cell injury. Methods RAEC cells were stimulated by LPS and given low-, medium- and high-dose (5, 10, 20 μmol/L) gentiopicroside for intervention. CCK-8 was used to detect cell proliferation; Flow cytometry was used to detect cell apoptosis; Reagent test kits were used to detect malondialdehyde level and superoxide dismutase (SOD) activity in cells; qRT-PCR was used to detect tumor necrosis factor-α (TNF-α), interleukin-6 (IL-6), and IL-1β mRNA levels in cells; Western blotting was used to detect p-p65 and cleaved Caspase-3 expressions in cells. RAEC cells were treated with nuclear factor-κB (NF-κB) signal activator to investigate the effect of NF-κB signal activator on gentiopicroside in improving RAEC cells damage. Results Compared with control group, cell activity in model group was significantly reduced (P<0.05), apoptosis rate was significantly increased (P<0.05), malondialdehyde level was significantly increased (P<0.05), SOD activity was significantly reduced (P<0.05), TNF-α, IL-6 and IL-1β mRNA levels were significantly increased (P<0.05), cleaved Caspase-3 and p-p65 expressions were significantly increased (P<0.05). Compared with model group, cell activity in gentiopicroside group was significantly increased (P<0.05), apoptosis rate was significantly reduced (P<0.05), malondialdehyde level was significantly reduced (P<0.05), SOD activity was significantly increased (P<0.05), TNF-α, IL-6 and IL-1β mRNA levels were significantly reduced (P<0.05), cleaved Caspase-3 and p-p65 expressions were significantly reduced (P<0.05). NF-κB signal activator significantly inhibited the effect of gentiopicroside in improving RAEC cells damage (P<0.05). Conclusion Gentiopicrin can inhibit LPS-induced apoptosis, oxidative damage and inflammatory factor expression in RAEC cells, which mechanism may be related to the inhibition of NF-κB signaling pathway.

R285.5

河南省科学技术计划项目（2018225019）