摘要
目的:探讨姜黄素(Curcumin)对肝纤维化体内外的作用和机制。方法:以人肝星状细胞LX-2为研究目标,CCK-8检测正常条件和TGF-β激活状态下LX-2细胞增殖情况;qRT-PCR和Western blot检测姜黄素对LX-2细胞CollagenⅠ、CollagenⅢ、α-SMA、p-smad2和p-65/NF-κB表达的影响;ELISA检测免疫炎性因子IL-10和IFN-γ的表达变化;CCl4诱导小鼠肝纤维化模型,100 mg/kg给药,检测小鼠给药前后血液中谷草转氨酶(AST)及谷丙转氨酶(ALT)、肌酐(Cr)及尿素氮(BUN)的水平变化;HE染色观察小鼠肝组织给药前后结构变化和损伤;马松染色观察小鼠肝组织给药前后纤维胶原表达的变化;Western blot检测小鼠肝组织给药前后α-SMA、p-smad2和p-65/NF-κB 表达的影响;羟脯氨酸试剂盒检测小鼠肝组织给药前后羟脯氨酸含量的变化。结果:CCK-8实验结果显示10、15、20、25 μmol/L 姜黄素对LX-2均有抑制作用,且20 μmol/L抑制作用最强(P<0.05);姜黄素显著抑制CollagenⅠ、CollagenⅢ、α-SMA、p-smad2和p-65以及免疫炎性因子IL-10和IFN-γ的表达(P<0.05);动物实验结果显示姜黄素可以有效改善CCL4诱导的肝组织损伤和肝功能损伤(P<0.05),减少肝组织内胶原的沉积(P<0.05),抑制肝组织内p-smad2和p-65/NF-κB表达(P<0.05)。结论:姜黄素通过抑制TGF-β/smad2信号通路从而抑制人肝星状细胞纤维化作用。
Objective:To explore the effect and mechanism of curcumin on liver fibrosis in vitro and in vivo.Methods:Using human liver stellate cells LX-2 as the research target,CCK-8 was used to detect the proliferation of LX-2 cells under normal conditions and TGF-β activation;qRT-PCR and Western blot were used to detect curcumin on LX-2 cells.Effects of curcumin on the expression of CollagenⅠ,CollagenⅢ,α-SMA,p-smad2 and p-65/NF-κB in LX-2 cells by qRT-PCR and Western blot.ELISA was used to detect changes in the expression of immunoinflammatory factors IL-10 and IFN-γ.CCl4-induced liver fibrosis in mice,100 mg/kg administration,the changes of aspartate aminotransferase(AST),alanine aminotransferase(ALT),creatinine(Cr) and urea nitrogen(BUN) levels in blood of mice before and after administration were measured.HE staining was used to observe the structure of mice liver tissue before and after administration changes and damages.Masson staining was used to observe the changes of fibrous collagen expression in mice before and after administration.Western blot was used to detect the effects of α-SMA,p-smad2 and p-65/NF-κB expression in mice before and after administration.The hydroxyproline kit was used to detect the changes of hydroxyproline content in the liver tissue of mice before and after administration.Results:CCK-8 results showed that 10,15,20,25 μmol/L curcumin inhibited LX-2,and 20 μmol/L had the strongest inhibitory effect(P<0.05),and 20 μmol/L produced cytotoxicity.Curcumin significantly inhibited the expression of CollagenⅠ,CollagenⅢ,α-SMA,p-smad2 and p-65 and immunoinflammatory factors IL-10 and IFN-γ(P<0.05).Animal experiments showed that curcumin could Effectively improve CCL4-induced liver injury and liver function damage(P<0.05),reduce collagen deposition in liver tissue(P<0.05),and inhibit the expression of p-smad2 and p-65/NF-κB in liver tissue(P<0.05).Conclusion:Curcumin inhibits fibrosis of human hepatic stellate cells by inhibiting the TGF-β/smad2 signaling pathway.
作者
王国泰
李京涛
魏海梁
闫曙光
李宁
WANG Guo-Tai;LI Jing-Tao;WEI Hai-Liang;YAN Shu-Guang;LI Ning(Hepatobiliary Surgery,Affiliated Hospital of Shaanxi University of Traditional Chinese Medicine,Xianyang 712000,China)
出处
《中国免疫学杂志》
CAS
CSCD
北大核心
2020年第4期422-427,共6页
Chinese Journal of Immunology
基金
陕西省科技厅科研基金(No.2016SF-234、No.2018KJXX-093)
陕西省中医药管理局科研基金(No.15-JC009)
陕西省“特支计划”区域发展人才项目(2017)
陕西中医药大学学科创新团队建设项目(No.2019-YL05)