丹参注射液对肝星状细胞的毒性及其作用机制研究  

Toxicity and mechanism of Salvianolate on hepatic stellate cell

作　　者：吴建辉[1] 裴晓丹[2] 邱晓燕[1] 刘向云[1] 蒋秀蓉[1] 刘桂明[1] 朱焰[1] 曹霖[1] 何桂林[1] 孙祖越[1] 

机构地区：[1]中国生育调节药物毒理检测中心上海市计划生育科学研究所药理毒理学研究室,上海200032 [2]辽宁大学环境科学系,沈阳110036

出　　处：《中药药理与临床》2006年第3期94-97,共4页Pharmacology and Clinics of Chinese Materia Medica

摘　　要：目的:探讨丹参注射液体外对肝星状细胞HSC-T6的毒性及其作用机制。方法:利用细胞培养技术,台盼兰法检测系列丹参溶液对HSC-T6生长的影响,G im sa染色观察细胞形态,MTT法检测丹参对细胞的生长抑制率,免疫组化检测血小板衍生生长因子(PDGF-BB)表达,生物法检测转化生长因子的表达。结果:①2.5至80μg/m l系列剂量丹参作用于HSC-T6 24、48和72h后,随着丹参剂量增大,HSC细胞存活率呈明显下降。②丹参作用48 h后,10μg/m l剂量部分细胞变成缩水球状,细胞胞浆变得不透明;20μg/m l剂量,脱壁细胞增多,细胞间隙增大,细胞很多呈缩水状;40μg/m l剂量多数细胞收缩成团块状,可见成片细胞脱壁;80μg/m l剂量细胞大量脱壁,活细胞很少。③HSC-T6经10至80μg/m l丹参作用24和48h后,490nm处测得OD值随剂量增大而降低,并随时间延长OD值减少。④免疫组化分析结果显示,与对照组相比,随着丹参剂量的增大,HSC-T6 PDGF-BB表达逐渐减弱,至20μg/m l差别已很明显(P<0.05)。⑤经10至40μg/m l丹参处理后HSC-T6 TGFβ1占总TGFβ的百分比降低。结论:丹参体外能够显著抑制HST-T6细胞增殖,其抑制机制是通过拮抗细胞因子PDGF-BB和TGFβ1的表达途径达到的。Aim： To explore the toxicity and mechanism of Danshen Injection on hepatic stellate cell in vitro. Methods： Effect of Danshen Injection on the proliferation of HSCs was detected by cell culture technology and trypan blue stain method, cell morphous was observed by Gimsa stain, Cell proliferation inhibition ratio was detected by methylthiazoly tetrazolium （MTT） method. The expression of PDGF and collegen was determined by immunohistochemical method. The expression of TGFβ1 was determined by biology method. Result： 1. After HSCs were cultivated with salvianolate for 24, 48 and 72 h, cell survival rate lower obviously by adding dose and has dose-dependent relationship. 2. After 48h, some cells that treated with 10μg/ml began shrinkage and turn into globular, endochylema becomes opacity. More cells of 20μg/ml group shed off and died, intercellular space widen and more began shrinkage, in 40μg/ml group, most cells became bolus and shed off culture flask, in 80μg/ml group, few cells alive. 3. After HSCs were cultivated with salvianolate for 24 and 48 h, OD value at 490 nm lower by adding dose and with time extension, OD value decreased. 4. The result of Immunohistochemical method demonstrates that the expression of PDGF-BB grew low when compared with control. Beyond 20μg/ml, difference was obvious（P 〈 0.05）. 5 The percentage of TGFβ1 content in total TGFβ lower after being treated with 10 to 40μg/ml salvianolate. Conclusion： Danshen Injection can obviously inhibit proliferation of HSC-T6, which mechanism can be achieved by inhibiting the expression of PDGF-BB and TGFβ1.

关 键 词：丹参注射液 肝星状细胞 细胞毒性 PDGF-BB 

分 类 号：R285[医药卫生—中药学]