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目的探讨降脂宁有效部位及其药效组分对人肝细胞Bel-7402胆固醇代谢的影响。方法体外培养人肝细胞株Bel-7402,待细胞长满80%后用无血清培养基饥饿细胞24 h,加入降脂宁有效部位及其药效组分和阿托伐也汀,高效液相色谱法(HPLC)和分光光度法检测给药24 h后细胞内胆固醇及胆汁酸含量;逆转录聚合酶链反应法(RT-PCR)检测细胞内低密度脂蛋白受体(LDL-R)和3羟基-3甲基-戊二酰辅酶A还原酶(HMG-CoAR)、7α-羟化酶(CYP7A1)mRNA表达水平。结果降脂宁有效部位及其药效组分和阿托伐他汀均能够降低细胞内胆固醇含量,增加细胞内胆汁酸含量,提高LDL-R及CYP7A1 mRNA表达水平,降脂宁有效部位及其药效组分高剂量组能够提高HMG-CoAR mRNA表达。结论降脂宁有效部位及其药效组分可以抑制胆固醇合成并促进其转化,其机制可能与影响细胞胆固醇代谢相关受体和酶mRNA表达水平有关。
Objective To investigate the effect of Jiangzhining active site and its components on the cholesterol metabolism of human hepatocellular Bel-7402 cells. Methods Human hepatocyte line Bel-7402 was cultured in vitro. When the cells were over 80%, the starved cells were treated with serum-free medium for 24 h. The effective fractions of Jiangzhining and its pharmacodynamic components and atorvastatin, Cholesterol and bile acid contents in the cells were detected by HPLC and spectrophotometry at 24 h after treatment. The levels of LDL-R and LDL-C were detected by reverse transcription-polymerase chain reaction (RT-PCR) 3 hydroxy-3-methyl-glutaryl coenzyme A reductase (HMG-CoAR), 7α-hydroxylase (CYP7A1) mRNA expression levels. Results The effective fractions of Jiangzhining, its medicinal components and atorvastatin all can reduce intracellular cholesterol content, increase intracellular bile acid content and increase the expression of LDL-R and CYP7A1 mRNA, High-dose group can increase HMG-CoAR mRNA expression. Conclusion The effective fractions of Jiangzhining and its pharmacodynamic components can inhibit the synthesis of cholesterol and promote its transformation. The mechanism may be related to the expression of receptors and enzymes related to cellular cholesterol metabolism.