二苯乙烯苷对TGF-β1诱导新生大鼠心肌成纤维细胞增殖和胶原合成的影响及其机制  

Effects and mechanisms of stilbene glucoside on the proliferation and collagen synthesis of cardiac fibroblasts stimulated by TGF-β1

作　　者：曾翼[1] 赵诚[1] 王霏[1] 朱秋燕[1] 许晓乐[1] 

机构地区：[1]南通大学药学院药理学教研室,南通226001

出　　处：《中国药科大学学报》2014年第3期362-367,共6页Journal of China Pharmaceutical University

基　　金：南通市科技计划资助项目(No.BK2012013);南通大学研究生科技创新计划资助项目(No.YKC12023;YKC13065)~~

摘　　要：探讨二苯乙烯苷（2，3，5，4＇-四羟基二苯乙烯-2-O-β-D葡糖苷，TSG）对转化生长因子β（TGF-β1）诱导的新生大鼠心肌成纤维细胞（CFB）增殖和胶原合成的影响，并研究其相关机制。用消化法培养新生SD大鼠CFB，建立TGF-β1诱导新生大鼠CFB纤维化模型。采用5-溴脱氧尿嘧啶核苷（BrdU）掺入法检测CFB增殖情况；乳酸脱氢酶（LDH）法检测药物对细胞毒性作用；羟脯氨酸测定检测CFB胶原含量；流式细胞仪测定细胞周期；Western blot法检测α-平滑肌肌动蛋白（α-SMA）、增殖细胞核抗原（PCNA）、Ⅰ型胶原（COLⅠ）、Ⅲ型胶原（COLⅢ）、p—Smad2、Smad2、p-Smad3、Smad3、p-ERK、ERK、p-P38、P38、p—JNK、JNK的蛋白表达。结果表明，终浓度为0．1—100μmol／L的TSG无明显细胞毒性作用；在一定浓度范围内，TSG可明显抑制TGF—β1诱导的CFB增殖和胶原合成，抑制细胞由G0／G1期向S期的转变，抑制TGF—β1诱导的CFB核内PC—NA的蛋白表达，降低α-SMA、COLI和COLⅢ的过度表达．并抑制TGF—β1诱导的Smad3、ERK和P38的磷酸化。因此推测一定浓度的TSG能抑制TGF—β1诱导的CFB增殖和胶原合成，其机制可能与干预Smad3、ERK和P38信号通路有关。The aim of this study was to investigate the effect of stilbene glucoside （2, 3, 4＇, 5-tetrahydroxystilbene-2-O-β-D glucoside, TSG） on the cell proliferation and collagen synthesis in cultured neonatal rat cardiac fibroblasts （CFBs） stimulated by transforming growth factor-β1 （TGF-β1） and to explore the underlying mechanisms. CFB were cultured from neonatal rats of 1-3 days and treated with TGF-β1. CFB proliferation was analyzed by 5-bromo-2-deoxyuridine （BrdU） incorporation assay. The synthesis of collagen was examined by measurement of the hydroxyproline concentration. Flow cytometry used to examined cell cycle of CFBs treated with TSG. The expressions of α-smooth muscle actin, proliferating cell nuclear antigen （PCNA）, collagen Ⅰ, collagen Ⅲ, p-Smad2, Smad2, p-Smad3, Smad3, p-ERK, ERK, p-P38, P38, p-JNK, JNK were measured by Western blot. The results of the present study showed that TSG （0. 1-100 μmol/L） to exerted no significant cytotoxicity on the cells. Within a concentration coverage, TSG inhibited CFB proliferation and collagen synthesis with inhibition of cell cycle transition from G0/G1 phase to S phase. Furthermore, TSG decreased the protein expressions of PCNA, α-smooth muscle actin, collagen Ⅰ , Collagen Ⅲ, p-Smad3, p-ERK and p-P38 stimulated by TGF-β1. Taken together, TSG significantly inhibited CFB proliferation and collagen synthesis stimulated by TGF-β1, though, at least in part, by suppression of Smad3, ERK and P38 activation.

关 键 词：二苯乙烯苷 心肌成纤维细胞 增殖 胶原合成 转化生长因子β1 

分 类 号：R965[医药卫生—药理学]