来氟米特对肝星状细胞增殖和胶原合成的影响  

The active metabolite of leflunomide A_(771726) inhibits proliferation and collagen synthesis of hepatic stellate cell

作　　者：姚宏伟[1] 李俊[2] 陈季强[1] 徐叔云[2] 

机构地区：[1]浙江大学医学院国家食品药品监督管理局浙江呼吸药物研究实验室,浙江杭州310031 [2]安徽医科大学临床药理研究所,安徽合肥230032

出　　处：《浙江大学学报（医学版）》2004年第6期515-518,528,共5页Journal of Zhejiang University(Medical Sciences)

基　　金：安徽省自然科学基金 (984 4 6 733)资助项目

摘　　要：目的 :研究来氟米特活性代谢物 A77172 6对肝星状细胞 (HSC)增殖和胶原合成的影响。方法 :用链蛋白酶和胶原酶原位肝灌注、Nycodenz密度离心法分离大鼠 HSC和 Kupffer细胞 ,制备 Kupffer培养上清液 (KCCM) ,并以不同浓度、不同培养时间的 CCl4损伤的 KCCM作为刺激因素观察了 A77172 6对 HSC增殖 (3 H- Td R掺入法 )和胶原合成 (3 H- Pro掺入法 )的影响。EL ISA法测定 KCCM内 TGF- β、TNF- α和 IL- 1含量。结果 :HSC和 Kupffer细胞得到较好的分离。CCl4损伤的 KCCM显著增加 HSC增殖和胶原合成 ,并且稀释度为 1∶ 2的 KCCM在培养 7d时 ,对 HSC增殖和胶原合成的刺激作用明显。 A77172 6(0 .0 0 1～ 10 μmol/ L)对 CCl4损伤的 KCCM刺激 HSC增殖和胶原合成有不同程度的抑制作用 ,并且 A77172 6(0 .0 0 1～ 10 μmol/ L)明显抑制 CCl4损伤 KCCM内升高的 TGF- β、TNF- α和 IL- 1水平。结论 :CCl4损伤的 KCCM可明显刺激 HSC增殖和胶原合成 ,而 A77172 6对此有明显的抑制作用 ,并且与其抑制致 HSC激活的因子如 TGF- β、TNF- α和 IL- 1分泌有关。Objective: To investigate the effect of the metabolite of leflunomide,A 771726 ,on proliferation and collagen synthesis of hepatic stellate cell (HSC). Methods: HSC and Kupffer cells were isolated from the rat liver by collagenase Ⅳ and pronase perfusion,and purified by density gradient separation.The effects of A 771726 on cell proliferation and collagen synthesis were examined by 3H thymidine and 3H proline incorporation assays,respectively.The TGF β,TNF α and IL 1 levels in Kupffer cell conditioned medium (KCCM) were determined by enzyme linked immunosorbent assay (ELISA). Results: HSC and Kupffer cells in rat liver were well separated.The KCCM of CCl 4 injured rats had significant stimulation effect on proliferation and collagen synthesis of HSC in primary culture. Addition of A 771726 (0.001~10 μmol/L)in HSC culture stimulated by KCCM significantly inhibited proli feration and collagen synthesis of HSC.Furthermore,the elevated TGF β,TNF α and IL 1 levels in KCCM of CCl 4 injured rats were significantly reduced in A 771726 treatment groups. Conclusion: A 771726 has markedly inhibitory effect on proliferation and collagen synthesis of HSC and secretion of TGF β,TNF α and IL 1 from Kupffer cells. [

关 键 词：来氟米特 A771726 肝星状细胞 KUPFFER细胞 细胞因子 

分 类 号：R967[医药卫生—药理学]