龙胆苦苷对脂多糖诱导血管内皮细胞损伤的作用及机制  

Effect and mechanism of gentiopicroside on vascular endothelial cell injury induced by lipopolysaccharide

作　　者：胡志良 池豪 丁水印 HU Zhi-liang;CHI Hao;DING Shui-yin(Cardiovascular Department of Zhumadian Central Hospital,Zhumadian 463000,China;Department of Cardiology,Zhengzhou Central Hospital,Zhengzhou 450000,China)

机构地区：[1]驻马店市中心医院心血管内科,河南驻马店463000 [2]郑州市中心医院心血管内科,河南郑州450000

出　　处：《中草药》2021年第10期3002-3008,共7页Chinese Traditional and Herbal Drugs

基　　金：河南省科学技术计划项目(2018225019)。

摘　　要：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、20μmol/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-1βmRNA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-1βm RNA表达水平显著升高(P<0.05),cleavedCaspase-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-1βmRNA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(5,10,20μmol/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-1βmRNA 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-1βmRNA 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-1βm RNA 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.

关 键 词：龙胆苦苷 血管内皮细胞 凋亡 炎性反应 氧化损伤 

分 类 号：R285.5[医药卫生—中药学]