丹参素对IL-1β刺激的大鼠肝星状细胞JNK、NF-κB信号通道的影响

Effect of danshensu on JNK and NF-κB signal transduction of rat hepatic stellate cells induced by interleukin-1β

目的观察丹参素对活化的大鼠肝星状细胞(HSC)氨基末端蛋白激酶(JNK)以及NF-κBP65表达的影响,探讨其抗肝纤维化作用的机制。方法分离大鼠原代HSC,MTT法检测丹参素对细胞的生长抑制,免疫细胞化学法观察Ⅲ型胶原合成,ELISA法观察Ⅲ型胶原分泌。AO/EB荧光染色观察HSC凋亡形态学变化,Annexin-V-FITC/PI联合流式细胞仪检测HSC的凋亡率,Western blotting观察丹参素对IL-1β刺激的HSCs内P-IκBα、NF-κBP65和JNK蛋白的表达。结果和结论丹参素可抑制活化的HSC增殖,降低胶原合成和分泌,促进HSC凋亡,并抑制IL-1β刺激的HSC中JNK的磷酸化及NF-κBp65的表达。

Objective To investigate the inhibitory effect of danshensu on c-Jun N-terminal kinase （JNK） and nuclear factor-kappa B （NF-κB） signaling in activated rat hepatic stellate cells （HSCs） and explore the mechanisms of danshensu for inhibiting hepatic fibrosis. Methods MTT colorimetric assay was used to detect the proliferation of rat HSCs treated with danshensu, and the apoptosis of the cells was analyzed with Annexin- V-FITC/PI and AO/EB staining. The expressions of P-IκBα, NF-κBP65 and JNK in HSCs stimulated by IL-1 β with subsequent danshensu treatment were observed by Western blotting. Type Ⅲ collagen in the medium of HSCs was detected by ELISA and immunocytochemistry. Results and Conclusions Danshensu inhibited the activation and proliferation of HSCs, and increased the apoptotic rate of HSCs and reduced the synthesis and secretion of type III collagen. Danshensu showed obvious inhibitory effect on JNK and P-IκB-α phosphorylation and NF-κBP65 expression in HSCs stimulated by IL-1 β. The mechanism of the actions ofdansgensu may be mediated by inhibition of JNK and NF-κB signal transduction.