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大黄素对脂多糖诱导的胆管上皮细胞炎症反应的影响

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目的 探讨大黄素对脂多糖(LPS)诱导的肝内胆管上皮细胞的影响及其作用机制.方法 用LPS方法建立肝内胆管上皮细胞炎症模型.将细胞随机分为对照组(正常培养),模型组(20 μg·mL-1 LPS),低、中、高剂量实验组(20μg·mL-1 LPS+5、10、20 μg·mL-1 大黄素)和 pcDNA-TLR4 组(转染pcDNA-TLR4质粒+20μg·mL-1 LPS+20 µg·mL-1 大黄素).用蛋白质印迹法检测Toll-样受体4(TLR4)、磷酸化核转录因子κB(p-NF-κB)、增殖细胞核抗原(PCNA)、上皮细胞钙黏蛋白(E-cadherin)的表达情况,用细胞计数试剂盒8(CCK-8)实验检测细胞增殖活性,用5-乙炔基-2'-脱氧尿苷(Edu)实验检测细胞增殖情况,用酶联免疫吸附试验法检测炎性因子水平.结果 对照组、模型组、低剂量实验组、中剂量实验组、高剂量实验组和pcDNA-TLR4组的TLR4 蛋白相对表达水平分别为 0.21±0.02、0.92±0.11、0.74±0.09、0.61±0.06、0.39±0.04和1.37±0.09,p-NF-κB蛋白相对表达水平分别为0.33±0.03、1.03±0.11、0.80±0.08、0.66±0.05、0.41±0.06 和 0.85±0.08,E-cadherin蛋白相对表达水平分别为 1.20±0.10、0.41±0.04、0.58±0.07、0.79±0.06、0.91±0.05 和 0.58±0.05,Edu 阳性细 胞率分 别 为(22.11±1.18)%、(51.78±2.35)%、(43.20±3.76)%、(33.19±3.58)%、(26.96±2.31)%和(41.40±2.74)%,白细胞介素-6(IL-6)水平分别为(9.79±1.35)、(38.69±3.99)、(29.80±3.41)、(25.48±1.50)、(15.63±0.91)和(27.75±3.61)pg·mL-1.对照组的上述指标与模型组比较,各剂量实验组的上述指标与模型组比较,pcDNA-TLR4组的上述指标与高剂量实验组比较,在统计学上差异均有统计学意义(均P<0.05).结论 大黄素可能通过抑制TLR4/NF-κB/c-myc通路抑制细胞过度增殖和上皮间质转化进而改善LPS诱导的胆管上皮细胞炎性反应.
Effects of emodin on lipopolysaccharide-induced inflammation of biliary epithelial cell and its mechanism
Objective To investigate the effect of emodin on lipopolysaccharide(LPS)-induced biliary epithelial cell and its mechanism.Methods Biliary epithelial cells were randomly divided into control group(normal culture),model group(20 μg·mL-1 LPS),experimental-L,-M,-H groups(20 μg·mL-1 LPS+5,10,20μg·mL-1 emodin),pcDNA-TLR4 group(20 μg·mL-1 LPS+20μg·mL-1 emodin+transfected pcDNA-TLR4 plasmid).Western blot assay was used to detect protein expression of Toll-like receptor 4(TLR4),phosphorylation-nuclear factor kappa B(p-NF-κB),proliferating cell nuclear antigen(PCNA),E-cadherin;cell counting kit 8(CCK-8)assay and 5-Ethynyl-2'-deoxyuridine(Edu)assay were used to detect cell proliferation;enzyme-linked immunosorbent assay(ELISA)assay was used to detect the expression level of inflammatory factors.Results The expression levels of TLR4 protein in control group,model group,experimental-L,-M,-H groups and pcDNA-TLR4 group were 0.21±0.02,0.92±0.11,0.74±0.09,0.61±0.06,0.39±0.04 and 1.37±0.09;the expression levels of p-NF-KB protein were 0.33±0.03,1.03±0.11,0.80±0.08,0.66±0.05,0.41±0.06 and 0.85±0.08;the expression levels of E-cadherin were 1.20±0.10,0.41±0.04,0.58±0.07,0.79±0.06,0.91±0.05 and 0.58±0.05;Edu positive cells rate were(22.11±1.18)%,(51.78±2.35)%,(43.20±3.76)%,(33.19±3.58)%,(26.96±2.31)%and(41.40±2.74)%;IL-6 levels were(9.79±1.35),(38.69±3.99),(29.80±3.41),(25.48±1.50),(15.63±0.91)and(27.75±3.61)pg·mL-1.Compared with control group and model group,compared with experimental-L,-M,-H groups and model group,pcDNA-TLR4 group was compared with experimental-H group,the differences of the above indexes were statistically significant(all P<0.05).Conclusion Emodin may inhibit cell hyperproliferation and EMT by inhibiting TLR4/NF-KB/c-myc pathway,thereby improving the LPS-induced inflammatory response of biliary epithelial cell.

emodinprimary biliary cholangitislipopolysaccharidebiliary epithelial cellepithelial mesenchymal transformation

闫凯、刘旭呈

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东南大学附属中大医院肝胆胰中心,江苏南京 210009

南京市第二医院 胃肠治疗中心,江苏 南京 210000

大黄素 原发性胆汁性胆管炎 脂多糖 胆管上皮细胞 上皮间质转化

2024

中国临床药理学杂志
中国药学会

中国临床药理学杂志

CSTPCD北大核心
影响因子:1.91
ISSN:1001-6821
年,卷(期):2024.40(3)
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