化浊解毒含药血清对大鼠肝星状细胞增殖及细胞因子的影响

目的:观察化浊解毒含药血清对体外培养活化的大鼠肝星状细胞增殖及活性的影响,探讨化浊解毒方治疗肝纤维化的作用机制.方法:不同剂量(分别含有生药30,60,120 g·L-1)的化浊解毒方,按0.01 mL·g-1体重SD大鼠灌胃给药,以制备含药血清；常规培养活化的肝星状细胞( HSC-T6),用不同浓度的含药血清进行干预,用MTT,ELISA,RT-PCR法分别检测肝星状细胞增殖、细胞上清液中Ⅰ型胶原的含量以及细胞中转化生长因子-β1(TGF-β1)mRNA基因的表达.结果:与正常血清组比较,化浊解毒方含药血清各浓度组均呈现出抑制HSC增殖的作用(P＜0.05),且呈剂量和时间依赖性.与正常血清组比较,化浊解毒方含药血清各个浓度组在作用48 h后,细胞上清液中的Ⅰ型胶原的含量均降低(P＜0.05),均能下调细胞中TGF-β1 mRNA的表达(P＜0.05).结论:化浊解毒方之所以能有效的治疗肝纤维化可能与化浊解毒方抑制肝星状细胞的增殖和活性有关.

Effects of Huazhuo Jiedu Recipe on Proliferation and Activity in Rat Cultured Hepatic Stellate Cells in vitro

Objective:To explore the effects of Huazhuo Jiedu Recipe(HZJD) on the proliferation and activity in rat hepatic Stellate cells(HSCs) in vitro.Method:The drug-containing serum of HZJD was prepared by feeding healthy SD rats at various doses(20,15,10 mL·kg-1·d-1) of HZJD,and used to incubate the HSCs cultured in vitro.The impact of drug-containing serum on HSCs proliferation was estimated by methyl thiazolyl tetrazolium(MTT)at 24,48 h and 72 h after by intervention of drug-containing serum;the content of collagen 1(COL I) was detected by enzyme linked immunosorbent assay(ELISA);the expression of transforming growth factor betal(TGF-β1) mRNA was measured by RT-PCR.Result: The proliferation of HSCs was inhibited significantly after intervention with various dose of HZJD drug-containing serum.With a concentration-dependent mannercompared with the normal serum control group(P<0.05).The content of COL I and the expression of TGF-β1 mRNA were degraded after intervened with HZJD drug-containing serum at various dose in HSCs(P<0.05).Conclusion: Drug-congtaining serum of HZJD could inhibit the proliferation and activity of HSCs in vitro.