丹参素对大鼠肝星状细胞氨基末端蛋白激酶信号转导的影响

目的 探讨丹参素抗肝纤维化作用的机制. 方法分离大鼠原代肝星状细胞(HSC),用0.0312、0.0625、0.1250、0.2500、0.5000、1.0000 mmol/I.不同浓度丹参素作用于HSC,四甲基偶氮唑黼法检测丹参素对HSC生长增殖的抑制作用;应用免疫细胞化学法观察Ⅰ型胶原合成,酶联免疫吸附法观察Ⅰ型胶原分泌.Western blot法观察丹参索对白细胞介素-1 β(IL-1 β)刺激的HSC内氨基末端蛋白激酶(JNK)蛋白及其磷酸化表达的影响.采用随机设计的方差分析,SNK-q进行统计学处理. 结果与0 mmol/L组比较,丹参素其他浓度组明显抑制了HSC增殖,并旱剂量依赖性.丹参素(0.0625～0.2500 mmol/L)对IL-1 β引起的HSC增殖具有明显的抑制作用,并呈剂量依赖性.0.25 mmol/L丹参素对正常培养的和经IL-1 β刺激的HSC作用24 h能下调Ⅰ型胶原的合成和分泌.IL-1 β能刺激ttSC中p-JNK的明显表达,丹参素能下凋亡IL-1 β诱导的ItSC中P-JNK的表达,但其对JNK表达水平没有明显影响. 结论丹参索可抑制正常传代培养的和经IL-1 β刺激的人鼠HSC增殖、Ⅰ型胶原合成与分泌,其机制可能与抑制JNK信号转导通路有关.

Effect of danshensu on activation of JNK pathway in hepatic stellate cells(HSCs) induced by IL-1 beta

Objective To investigate the inhibitory effect of danshensu on the activation of JNK signaling in rat hepatic stellate cells (HSCs) induced by IL-1 β. Methods The proliferation of primary rat HSCs treated with different concentration of Danshensu was checked by MTT colorimetric assay. The expres-sion and phosphorylation of JNK and P-JNK was detected by western blotting. Synthesis and secretion of collagen Ⅰ were detected by the quantitative immunocytochemical assay and ELISA. Results Danshensu inhibited the proliferation of HSCs in a dose-dependent manner. At the concentration of 0.0625 to 0.25 mmol/L, Danshensu significantly repressed the proliferation of HSC induced by IL-115 (P < 0.05). Synthesis and secretion of Type β collagen was significantly decreased 24 hours after 0.25 mmol/L Danshensu treatment (P < 0.01). The phosphorylation of JNK induced by IL-1 β was significantly inhibited by Danshensu treatment (P < 0.05), however, the expression of JNK was not regulated by Danshensu. Conelutions Danshensu represses the activation and proliferation of HSCs, and inhibits the synthesis and secretion of Type β collagen, possibly via the repression of the JNK signal transduction.