组分复方“CKJ”药物血清对大鼠原代肝星状细胞活化和转化生长因子β_1及其受体的影响  

Effect of CKJ Recipe Containing Serum on Activation of Rat Primary Hepatic Stellate Cells, TGF-β_1 and Its Receptors

作　　者：陈亮[1] 冯琴[1,2] 彭景华[1,2] 刘林[1] 梁春耕[1] 海亚美 胡义扬[1,3,4,2] 

机构地区：[1]上海中医药大学附属曙光医院上海中医药大学肝病研究所,上海201203 [2]上海中医药大学肝肾疾病病证教育部重点实验室,上海201203 [3]上海市高校中医内科学E-研究院,上海201203 [4]上海市中医临床重点实验室,上海201203

出　　处：《中国中西医结合杂志》2015年第2期210-215,共6页Chinese Journal of Integrated Traditional and Western Medicine

基　　金：国家科技重大专项资助项目(No.2012ZX09103-201-048)

摘　　要：目的探讨由虫草多糖、苦杏仁苷、绞股蓝总皂苷组成的"CKJ方"药物血清对大鼠原代肝星状细胞(primary hepatic stellate cells,r HSCs)活化的影响及相关药理作用机制。方法分离并培养r HSCs,将培养4天的r HSCs分为正常组、模型组及CKJ组。模型组及CKJ组以2.5 ng/m L转化生长因子β1(TGF-β1)刺激造模24 h,正常组以不含TGF-β1的基质液(无血清的DMEM)作为对照。药物组以5%CKJ方药物血清继续孵育24 h,正常组及模型组以5%空白血清孵育24 h。采用细胞高内涵筛选技术(HCS)免疫荧光法及Western blot检测r HSCsα-平滑肌肌动蛋白(α-SMA)表达;实时荧光定量聚合酶链式反应(qRT-PCR)检测α-SMA、Ⅰ型胶原蛋白(Col-Ⅰ)、血小板源性生长因子β受体(PDGF-βR)、TGF-β1、转化生长因子β受体1(TGF-βR1)、转化生长因子β受体2(TGF-βR2)mRNA的表达水平。结果与正常组比较,模型组r HSCsα-SMA蛋白表达及α-SMA、Col-Ⅰ、PDGF-βR、TGF-β1、TGF-βR1、TGF-βR2 mRNA表达水平明显升高(P<0.05,P<0.01);与模型组比较,5%CKJ药物血清组r HSCsα-SMA蛋白表达及α-SMA、Col-Ⅰ、PDGF-βR、TGF-β1、TGF-βR1、TGF-βR2 mRNA表达水平明显降低(P<0.05,P<0.01)。结论组分复方"CKJ"药物血清可降低大鼠r HSCsα-SMA的蛋白表达,其主要作用机理可能与抑制TGF-β1及其相关受体有关。Objective To observe the effect of CKJ Recipe （consisting of Cordyceps sinensis polysaccharide, amygdaloside, and gypenosides） containing serum on the activation of rat primary hepatic stellate cells （rHSCs） and to explore its pharmacological mechanism. Methods rHSCs were isolated form liver and cultured for four days. Then they were divided into the normal control group, the model group, and the CKJ group, rHSCs in the model group and the CKJ group were treated with 2.5 ng/mL transforming growth factor β1 （TGF-β1） in serum-free DMEM for 24 h. Serum free DMEM （containing no TGF-β1 ） was taken as the control for the normal control group, rHSCs in the CKJ group were treated with 5% CKJ-containing serum for 24 h. rHSCs in the other two groups were treated with 5% blank serum for 24 h. The protein expression level of a-smooth muscle actin （a-SMA） was determined using high through-put screening （HCS） and Western blot. mRNA expression levels of （x-SMA, collagen I （Col-I）, platelet-de- rived growth factor receptor β （PDGF-IBR）, TGF-I31, transforming growth factor β receptor 1 （TGF-βR1）, and transforming growth factor β receptor 2 （TGF-β R2 ） were detected using quantitative RT-PCR. Results Compared with the normal control group, the protein expression level of ot-SMA, mRNA expres- sion levels of （x-SMA, Col-I, PDGF-βR, TGF-β1, TGF-βR1, and TGF-βR2 significantly increased in the model group （P 〈0.05, P 〈0.01 ）. Compared with the model group, the protein expression level of ot-SMA, mRNA expression levels of a-SMA, Col-I, PDGF-βR, TGF-β1, TGF-β1, and TGF-β R2 significantly decreased in the CKJ group （P 〈0. 05, P 〈0.01 ）. Conclusion CKJ containing serum could inhibit the protein expression level of a-SMA, which was probably related with inhibiting TGF-β1 and its related receptors.

关 键 词：CKJ方 转化生长因子Β1 原代肝星状细胞 

分 类 号：R285.5[医药卫生—中药学]