丹酚酸B对MDA刺激的肝星状细胞增殖的抑制作用  

Inhibitory effect of salvianolic acid B on malondialdehyde-stimulated proliferation of rat hepatic stellate cells in vitro

作　　者：王晓玲[1] 刘平[2] 崔云华[3] 胡旭东[1] 李伯勤[4] 应馨萍[4] 

机构地区：[1]上海中医药大学生物教研室,上海市201203 [2]上海中医药大学肝病研究所,上海市201203 [3]上海中医药大学针灸经络研究所,上海市201203 [4]上海中医药大学教学实验中心,上海市201203

出　　处：《世界华人消化杂志》2006年第5期476-480,共5页World Chinese Journal of Digestology

基　　金：国家自然科学基金青年基金资助项目;No.30000230~~

摘　　要：目的:研究丹酚酸B的抗氧化性对大鼠肝星状细胞(HSC)的影响．方法:采用原位灌注法消化大鼠肝脏,108 g／L Nycodenz密度梯度离心,分离肝星状细胞,分别以不同浓度MDA／SAB处理细胞,MTT法观察细胞的增殖能力,2’,7’-二氯二氢荧光素 (DCFH)掺入反映细胞内氧化水平,Western blot检测增殖细胞核抗原(PCNA)蛋白含量, 免疫组化法检测血小板衍生的生长因子受体 (PDGFR)含量．结果:MDA刺激后,细胞增殖明显增强, MTT结果显示,吸光度与对照组相比显著升高(0．253±0．016 vs 0．213±0．004,P<0．05), 而1 μmol／L SAB(0．182±0．006,P<0．01)和 10 μmol／L SAB(0．179±0．006,P<0．01)均可以显著抑制MDA刺激的HSC增殖．Western blot显示,PCNA蛋白在MDA刺激后明显增加(1．72±0．026 vs 1．223±0．025,P<0．01),而 1和10 μmol／L SAB可显著抑制PCNA蛋白表达的升高(分别是1．080±0．040和1．066± 0．025,P<0．01)．MDA可明显刺激细胞PDGF 受体的表达(5．5±0．653 vs 对照组3．3±0．616, P<0．01),提高HSC细胞内氧化水平(荧光强度: 4．721±0．385 vs 对照组2．413±0．662,P<0．01), 10 μmol／L SAB则可抑制PDGF受体的表达(2．723±0．326)和降低细胞内的氧化水平 (3．324±0．264)(P<0．01)．结论:丹酚酸B可通过影响PDGF信号通路而抑制体外培养HSC的增殖,且这种抑制作用与丹酚酸B的抗氧化作用有关．AIM： To investigate the inhibitory effects of salvianolic acid B （SAB） on the proliferation of rat hepatic steUate ceils stimulated by malondialdehyde （MDA）. METHODS： Hepatic stellate cells were derived from normal rat by in situ perfusion, digestion and density gradient centrifugation with 108 g/L Nycodenz. Then the cells were stimulated by MDA （200 μmol/L） for 2 h, and then treated with various concentrations of SAB （1 and 10μmol/L）. The proliferation of cells was assayed by MTT incorporation, and the level of intracellular oxidation was reflected by 2＇, 7＇-dichlorodihydrofluorescein （DCFH）. The protein expression of proliferating cell nuclear antigen （PCNA） was determined by Western blot and the expression of platelet-derived growth factor receptor （PDGFR） was detected by immunohis-tochemistry. RESULTS： After MDA stimulation, the optical density by MTT was significantly increased in comparison with that of control cells （0.253 ± 0.016 vs 0.213 ± 0.004, P 〈 0.05）. However, after treatment with SAB, the optical density was markedly decreased （1 μmol/L： 0.182 ± 0.006, P 〈 0.01; 10μmol/L： 0.179 ± 0.006, P 〈 0.01）. The protein expression of PCNA was notably elevated after MDA stimulation （1.72 ± 0.026 vs 1.223 ± 0.025, P 〈 0.01）, while it was significantly downregulated by 1 and 10μmol/L SAB （1.080 ± 0.040 and 1.066 ± 0.025, respectively, P 〈 0.01）. MDA increased the expression of PDGFRβ （optical density： 5.5 ± 0.653 vs control, 3.3 ± 0.616, P 〈 0.01） and intracellular oxidative level （fluorescence intensity： 4.721 ± 0.385 vs control, 2.413 ± 0.662, P 〈 0.01）, but 10μmol/L SAB suppressed this effect （2.723 ± 0.326, 3.324 ± 0.264 vs MDA, P 〈 0.01）. CONCLUSION： SAB can inhibit MDA-stimulated proliferation of rat hepatic steUate cells in vitro by affecting PDGF signal transduction, which is associated with the antioxidant effect of SAB.

关 键 词：丹酚酸B 肝星状细胞 增殖周期 丙二醛 

分 类 号：R575.2[医药卫生—消化系统]