摘要
目的探讨扶肝化纤汤对肝星状HSC-T6细胞活化增殖及c-Jun氨基末端激酶(c-Jun N-terminal kinase,JNK)信号通路的影响。方法SD大鼠ig扶肝化纤汤制备含药血清,转化生长因子-β1(transforming growth factor-β1,TGF-β1)诱导HSC-T6细胞活化,设置对照组(10%胎牛血清)、模型组(15%正常血清)、扶肝化纤汤含药血清组(15%含药血清)、JNK抑制剂组(15%正常血清、20μmol/L SP600125),干预24 h。CCK-8法检测各组HSC-T6细胞增殖情况;ELISA法检测细胞上清液中白细胞介素-1β(interleukin-1β,IL-1β)、IL-6和肿瘤坏死因子-α(tumor necrosis factor-α,TNF-α)水平;Western blotting检测p-JNK和α-平滑肌肌动蛋白(α-smooth muscle actin,α-SMA)表达;qRT-PCR检测JNK和α-SMA m RNA表达;细胞免疫荧光检测α-SMA表达。结果TGF-β1诱导24 h后,HSC-T6细胞变长,胞体变大,胞质内折光颗粒减少,部分细胞融合成片状。与模型组比较,各给药组均可明显抑制HSC-T6增殖(P<0.05),显著抑制炎症因子IL-1β、IL-6、TNF-α的分泌(P<0.05),下调p-JNK、α-SMA蛋白表达及JNK、α-SMA mRNA表达水平(P<0.05),扶肝化纤汤含药血清与JNK抑制剂作用相当。结论扶肝化纤汤可抑制HSC细胞活化增殖,减少活化HSC-T6细胞JNK、α-SMA m RNA表达,降低pJNK、α-SMA蛋白表达水平。抑制JNK信号通路激活可能是扶肝化纤汤抗肺纤维化的另一种作用机制。
Objective To explore the effect of Fugan Huaxian Decoction(扶肝化纤汤)on activation and proliferation of hepatic stellate HSC-T6 cells and the signal pathway of c-Jun N-terminal kinase(JNK).Methods SD rats were ig Fugan Huaxian Decoction to prepare medicated serum,and HSC-T6 cells were activated by transforming growth factor-β1(TGF-β1).The rats were divided into control group(10%fetal bovine serum),model group(15% normal serum),Fugan Huaxian Decoction medicated serum group(15%medicated serum)and JNK inhibitor group(15% normal serum and 20μmol/L SP600125).The proliferation of HSC-T6 cells in each group was detected by CCK-8 method.The levels of interleukin-1β(IL-1β),IL-6 and tumor necrosis factor-α(TNF-α)in cell supernatant were detected by ELISA.Western blotting was used to detect the expressions of p-JNK and α-smooth muscle actin(α-SMA).The expressions of JNK andα-SMA mRNA was detected by qRT-PCR.The expression ofα-SMA was detected by cell immunofluorescence.Results After 24 h of TGF-β1 induction,HSC-T6 cells became longer,the cell body became larger,the refractive particles in cytoplasm was decreased,and some cells merged into sheets.Compared with model group,each drug group significantly inhibited the proliferation of HSC-T6(P<0.05),significantly inhibited the secretion of inflammatory factors IL-1β,IL-6 and TNF-α(P<0.05),and down-regulated p-JNK andα-SMA protein and JNK,α-SMA mRNA expressions(P<0.05).Fugan Huaxian Decoction medicated serum had the same effect as JNK inhibitor.Conclusion Fugan Huaxian Decoction can inhibit the activation and proliferation of HSC cells,reduce the expressions of JNK and α-SMA mRNA in activated HSC-T6 cells,and reduce the expression levels of p-JNK and α-SMA protein.Inhibition of JNK signaling pathway activation may be another mechanism of anti-hepatic fibrosis of Fugan Huaxian Decoction.
作者
肖政华
李婷婷
石以石则
杨庆万
李珊珊
XIAO Zheng-hua;LI Ting-ting;SHIYI Shi-ze;YANG Qing-wan;LI Shan-shan(Department of Gastroenterology,The Second Affiliated Hospital of Guizhou University of Traditional Chinese Medicine,Guiyang 550001,China;Guizhou University of Traditional Chinese Medicine,Guiyang 550025,China;The First People’s Hospital of Lanzhou New Area,Lanzhou 730300,China;Meishan Hospital of Traditional Chinese Medicine,Meishan 620000,China)
出处
《中草药》
CAS
CSCD
北大核心
2023年第16期5283-5288,共6页
Chinese Traditional and Herbal Drugs
基金
国家自然科学基金资助项目(81660790)
青年岐黄学者项目(国中医药人教发[2020]7号)
贵州省高等学校中西医结合防治疾病转化医学重点实验室项目(黔教技[2023]017号)。
