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、20 μmol/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 mRNA表达；细胞免疫荧光检测α-SMA表达。结果 TGF-b1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 mRNA表达，降低p-JNK、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 20 μmol/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 and α-SMA mRNA was detected by qRT-PCR. The expression of α-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 and α-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.

R285.5

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）