大黄素对脂多糖诱导的胆管上皮细胞炎症反应的影响  

Effects of emodin on lipopolysaccharide-induced inflammation of biliary epithelial cell and its mechanism

作　　者：闫凯 刘旭呈 YAN Kai;LIU Xu-cheng(Hepatobiliary and Pancreatic Center,Zhongda Hospital Affiliated to Southeast University,Nanjing 210009,Jiangsu Province,China;Gastrointestinal Treatment Center,Nanjing Second Hospital,Nanjing 210000,Jiangsu Province,China)

机构地区：[1]东南大学附属中大医院肝胆胰中心,江苏南京210009 [2]南京市第二医院胃肠治疗中心,江苏南京210000

出　　处：《中国临床药理学杂志》2024年第3期368-372,共5页The Chinese Journal of Clinical Pharmacology

摘　　要：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(20μg·mL^(-1) LPS),低、中、高剂量实验组(20μg·mL^(-1) LPS+5、10、20μg·mL^(-1)大黄素)和pcDNA-TLR4组(转染pcDNA-TLR4质粒+20μg·mL^(-1) LPS+20μg·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。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+5,10,20 μg·mL^(-1) emodin),pcDNA-TLR4 group(20 μg·mL^(-1) LPS+20 μg·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 pc DNA-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-κB 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·m L^(-1).Compared with control group and model group,compared with experimental-L,-M,-H groups and model group,pc DNA-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-κB/c-myc pathway,thereby improving the LPS-induced inflammatory response of biliary epithelial cell.

关 键 词：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 

分 类 号：R28[医药卫生—中药学]