丹参酚酸B盐对内皮素诱导的人肝星状细胞收缩的抑制作用及机制研究  

Effect and Mechanism of Salianic-acid B on Inhibiting Endothelin-1-induced Contraction of Human Hepatic Stellate Cells

作　　者：张杰[1,2] 张文炜[1,2] 徐列明[1,2,3] 

机构地区：[1]上海中医药大学附属曙光医院,上海201203 [2]上海市中医药研究院肝病研究所 [3]上海中医药大学肝肾疾病病证教育部重点实验室

出　　处：《中国中西医结合杂志》2009年第1期60-64,共5页Chinese Journal of Integrated Traditional and Western Medicine

基　　金：国家自然科学基金资助项目(No.30672489);;上海市重点学科建设资助项目(No.Y0302)

摘　　要：目的以培养的人肝星状细胞(HSC)为研究对象,观察内皮素-1(ET-1)对HSC的收缩作用及丹参酚酸B盐(SA-B)对ET-1的抑制作用,并探讨SA-B抑制HSC收缩的作用环节和可能作用机制。方法采用酶消化、Nycodenz密度梯度离心的方法分离人HSC,以胶原凝胶收缩法观察ET-1对于传代HSC的收缩作用,以及低、中、高3种剂量的SA-B对其不同的干预作用;将ET-1、SA-B直接添加于传代HSC的无血清培养液中,以激光共聚焦显微镜技术观测HSC内钙离子浓度的变化。结果胶原凝胶收缩实验显示,ET-1可诱导HSC收缩(P<0.01);3种剂量的SA-B可明显抑制ET-1诱导的HSC收缩(均P<0.01);加入ET-1后,HSC形态学改变明显,细胞数量减少,但SA-B对这些改变也有抑制。激光共聚焦显微镜下观察,ET-1可刺激细胞内钙离子短暂迅速增加,加入SA-B后,该作用被明显抑制。结论SA-B能显著抑制ET-1诱导的HSC收缩,其抑制收缩的机制与降低HSC内的钙离子浓度有关。SA-B的抑制HSC收缩作用可能是其抗肝纤维化及门脉高压的机制之一。Objective To observe the contraction effect of endothelin-1 （ ET-1 ） on human hepatic stellate cells （HSCs） and the inhibition of salianic-acid B （SA-B） on ET-1, to explore the acting link and the possible mechanism. Methods HSC were isolated from human normal liver tissue by enzyme digestion and Nycondenz density gradient centrifugation. The contraction of ET-1 on passage HSCs and the intervention of SA-B with three doses （low-, middle-, and high-） on the contraction were observed by collagen gel contraction. ET-1 and SA-B were directly added to the serum-free medium of HSCs, then calcium ion concentration was detected by laser scanning confocal microscope. Results Collagen gel contraction experiments showed that ET-1 could induce the contraction of HSC directly （ P 〈0.01 ）. Three doses of SA-B significantly inhibited the contraction effects of ET- 1 on HSCs（ all P 〈0.01 ）. After adding the ET-1, HSCs morphology changed obviously with the number of cells decreased. However, SA-B inhibited the changes. Laser scanning confocal microscope experiments revealed that ET-1 stimulated the transiently rapid increase of intracellular calcium ion concentration, and the effects was obviously inhibited when SA-B was added. Conclusions SA-B could inhibit the contraction of HSCs induced by ET-1, and its mechanism might be related to the lowing of free calcium ion concentration in HSCs. This anti-con- traction effect of SA-B is perhaps one of the mechanisms of its anti-fibrosis and anti-portal hypertension effects.

关 键 词：丹参酚酸B盐 肝纤维化 肝星状细胞 细胞收缩 内皮素-1 游离钙离子 

分 类 号：R285.5[医药卫生—中药学]