拆方益肝康不同药物血清对肝星状细胞胶原代谢及TGF-β1的影响  

Seropharmacological effects of Yigankang Decoction and its separated recipe on collagen metabolism and transforming growth factorbeta 1 expression in hepatic stellate cells

作　　者：房澍名[1] 姚冬梅[1] 张晓岚[1] 姚希贤[1] 

机构地区：[1]河北医科大学第二医院消化科,河北省消化病重点实验室,河北省消化病研究所,河北省石家庄市050000

出　　处：《世界华人消化杂志》2009年第27期2773-2777,共5页World Chinese Journal of Digestology

基　　金：国家自然科学基金资助项目;No.30872513;河北省中医药管理局基金资助项目;No.200335~~

摘　　要：目的:探讨益肝康抗肝纤维化作用机制、配伍意义,并采用对比血清药理学方法探讨更为科学的中药研究方法.方法:分别制备益肝康及其拆方-丹参小复方正常大鼠(A组,A1组:正常大鼠血清对照组;A2组:正常大鼠丹参小复方血清组;A3组:正常大鼠益肝康血清组)与肝纤维化大鼠(B组,B1组:肝纤维化大鼠血清对照组;B2组:肝纤维化大鼠丹参小复方血清组;B3组:肝纤维化大鼠益肝康血清组)药物血清,温育体外培养的HSC,3H-脯氨酸掺入法检测胶原合成,Western blot技术检测TGF-β1蛋白表达,RT-PCR技术检测TGF-β1mRNA表达.结果:益肝康及丹参小复方正常大鼠与肝纤维化大鼠药物血清均可以抑制HSC胶原合成(100mL/L时A组:2275.00±114.30cpm,2401.87±108.50cpm vs2963.62±128.01cpm;B组:2205.31±108.97cpm,2462.70±177.02cpm vs3179.12±223.34cpm;200mL时A组:2372.96±123.3cpm vs2515.37±98.25cpm vs3209.38±110.75cpm;B组:2394.29±150.50cpm vs2611.25±126.05cpm vs3490.46±183.16cpm,P<0.05或0.01),100mL时降低TGF-β1基因及蛋白表达(均P<0.01).2组在胶原合成、TGF-β1基因及蛋白表达益肝康组作用优于丹参小复方(均P<0.01),而益肝康组与丹参小复方组比较则B组血清作用优于A组(益肝康组:TGF-β1基因:0.356±0.032vs0.568±0.028;TGF-β1蛋白:0.458±0.009vs0.639±0.102;丹参小复方组:0.601±0.047vs0.810±0.051;0.612±0.126vs0.860±0.138,P<0.05或P<0.01).结论:益肝康及丹参小复方均可抑制HSC胶原合成,其主要机制之一是抑制TGF-β1基因和蛋白表达,益肝康更具配伍意义,肝纤维化大鼠药物血清药效优于正常大鼠血清药效.AIM：To investigate the seropharmacological effects of Yigankang Decoction and its separated recipe on collagen metabolism and transforming growth factor-beta 1 （TGF-β1） expression in hepatic stellate cells （HSCs） and explore the antifibrotic mechanism of Yigankang Decoction.METHODS：Normal rats and rats with carbon tetrachloride （CCl4）-induced liver fibrosis were administered Yigankang Decoction and its separated recipe （containing radix salvia militiorrhizae,astragalus and angelica sinensis）,respectively,to prepare medicated sera. After medicated rat sera were incubated with subcultured HSCs,the synthesis of collagen was evaluated by 3H-proline incorporation assay,and the expression of TGF-β1 mRNA and protein was detected by reverse transcription-polymerase chain reaction （RT-PCR） and Western blot,respectively. RESULTS：All kinds of medicated rat sera prepared in the study could inhibit 3H-proline incorporation at a concentration of 100 or 200 mL/L （normal rat sera at 100 mL/L：2275.00 ± 114.30 cpm vs 2401.87 ± 108.50 cpm vs 2963.62 ± 128.01 cpm; fibrotic rat sera at 100 mL/L：2205.31 ± 108.97 cpm vs 2462.70 ± 177.02 cpm vs 3179.12 ± 223.34 cpm; normal rat sera at 200 mL/L：372.96 ± 123.3 cpm vs 2515.37 ± 98.25 cpm vs 3209.38 ± 110.75 cpm; fibrotic rat sera at 200 mL/L：2394.29 ± 150.50 cpm vs 2611.25 ± 126.05 cpm vs 3490.46 ± 183.16 cpm; all P〈0.05 or 0.01）,and decrease the expression of TGF-β1 mRNA and protein at a concentration of 100 mL/L （all P〈0.01）. The sera from rats medicated with Yigankang Decoction had more significant inhibitory effects on 3H-proline incorporation and TGF-β1 expression than those from rats medicated with the separated recipe （all P〈0.01）. The sera from medicated fibrotic rats were superior to those from medicated normal rats in the downregulation of TGF-β1 expression （Yigankang Decoction：TGF-β1 mRNA,0.356 ± 0.032 vs 0.568 ± 0.028; TGF-β1 protein,0.458 ± 0.009 vs 0.639 ± 0.102; the separated recipe：TGF-β1 mRNA

关 键 词：益肝康 肝纤维化 肝星状细胞 转化生长因子beta-1 血清药理学 

分 类 号：R285.5[医药卫生—中药学]