化淤祛痰方药含药血清对HepG2细胞胆固醇代谢相关基因表达的影响  

Quantification of Rubusoside in Preparations of Rubus Suavissimus by RSLC-DAD

作　　者：贾连群[1] 杨关林[1] 罗莹[2] 任路[1] 冯峻屹[1] 崔勇[1] 陈阳[3] 

机构地区：[1]辽宁中医药大学,辽宁沈阳110032 [2]大连医科大学,辽宁大连116044 [3]辽宁省沈阳市第六人民医院,辽宁沈阳110006

出　　处：《时珍国医国药》2012年第1期127-129,共3页Lishizhen Medicine and Materia Medica Research

基　　金：辽宁省科技厅博士启动基金项目(No.20091052);辽宁省教育厅创新团队基金项目(No.2009T068);中国博士后科学基金项目(No.20090451279)

摘　　要：目的探讨化淤祛痰方药含药血清对HepG2细胞胆固醇摄取和合成相关基因表达的影响。方法体外培养HepG2细胞,随机分为5组,①正常对照组:仅用1640完全培养基培养,不加任何药物。②ox-LDL刺激组:培养液中加入终浓度为50μg/ml ox-LDL;③含药血清低剂量组;④含药血清中剂量组;⑤含药血清高剂量组。③～⑤组细胞分别使用5%、10%、20%含药血清预处理24 h后加入终浓度为50μg/ml ox-LDL,各组细胞培养24 h后进行各项指标测定。采用MTT法检测细胞活性;胆固醇氧化酶终点法检测HepG2细胞总胆固醇含量;反转录-聚合酶链反应(RT-PCR)定量分析3-羟基-3-甲基戊二酸单酰辅酶A还原酶(HMGCR)和B类Ⅰ型清道夫受体(SR-BⅠ)基因表达的变化。结果50μg/ml ox-LDL作用于HepG2细胞使其细胞活力显著下降,细胞总胆固醇含量显著升高,而化淤祛痰方药含药血清可以提高细胞的存活率,降低细胞内胆固醇含量。RT-PCR结果表明经ox-LDL诱导HepG2细胞HMGCR mRNA表达显著减少,而SR-BⅠmRNA表达显著增加。不同浓度的化淤祛痰方药含药血清均能显著降低HMGCR mRNA水平,显著升高SR-BⅠmRNA水平。结论化淤祛痰方药可通过降低HMGCR水平和升高SR-BⅠ水平从而促进胆固醇逆向转运,这可能是其调控脂质代谢发挥抗AS的作用机制之一。Objective To establish a rapid separation liquid chromatography with Diode Array Detecter（RSLC-DAD） method for quantification of rubusoside in Rubus Suavissimus preparations. Methods The separation was performed on a Dionex RSLC Acclaim 120 C18（50 mm×2.1mm,2.2μm） column with the mobile phase consisted of acetonitrile-water（36∶64） at the flow rate of 0.3 ml/min.The detection wavelength was set at 205nm,and the column temperature was kept constantly at 40 ℃.The injection volume was 2 μl. Results The linear range was 0.03~0.6μg for rubusoside.The average recovery was 99.1% with RSD of 0.97%（n=9）.The content range of rubusoside were 0 ~ 63.79 mg/g in 21 Sweet tea preparations from China or Japan. Conclusion The method is simple,rapid,accurate,and helpful for quantification of rubusoside in R.Suavissimus preparations.

关 键 词：化淤祛痰 动脉粥样硬化 胆固醇代谢 

分 类 号：R285.5[医药卫生—中药学]