摘要
目的探讨慢性乙型肝炎病毒(HBV)感染对人肝脏细胞色素酶3A4(CYP3A4)的影响,为慢性 HBV 感染患者安全用药提供指导。方法收集慢性 HBV 感染患者和正常人的肝组织标本,匀浆后以差速离心法制备成微粒体。以睾丸酮为探针底物,用高效液相色谱(HPLC)方法建立体外检测 CYP3A4酶代谢活性的技术平台,检测两组肝组织样本中 CYP3A4的酶活性。以 Western 印迹法测定了两组肝组织样本中 CYP3A4蛋白的表达差异。结果 HPLC 检测显示,反映酶活力的指标V_(max):慢性 HBV 感染组为(493±297)pmol·min^(-1)·mg^(-1),正常对照组为(741±189)pmol·min^(-1)·mg^(-1),两组差异有统计学意义(P=0.03),而酶系特征性常数 Km 值两组分别为:(0.142±0.057)μmol/L,(0.121±0.024)μmol/L,差异无统计学意义(P=0.103)。Western 印迹测定 CYP3A4蛋白表达两组差异有统计学意义(P=0.003),慢性 HBV 感染组 CYP3A4蛋白表达为3.0±1.6,低于正常组的表达量(5.7±1.5)。各样本酶活性和蛋白表达的相关系数为0.7683(P<0.01),表明酶活性与蛋白表达呈现良好的相关性。结论慢性 HBV 感染可降低肝脏 CYP3A4酶蛋白的表达,导致酶活性下降,但是对酶的结构未产生影响。提示慢性 HBV 感染者在使用由 CYP3A4代谢的药物时,要重视由于代谢变化导致的副作用。
Objective To study the effect of HBV on human hepatic cytoehrome P450 3A4 （CYP3A4） in patients with chronic HBV infection. Methods Liver tissue samples were obtained from 21 patients undergoing hepatic surgery with （ n = 10 ） or without （ n = 11 ） chronic HBV infection and were homogenized, then hepatic microsomes were separated from the homogenate using a differential centrifugation method. The activity of CYP3A4 was determined by HPLC, and the expression of CYP3A4 protein was determined by Western-blotting. Results The Vmax of CYP3A4 in patients with chronic HBV infection was 493±297 μmol/min/mg , significantly reduced （P =0. 03） comparing to the normal control group： 741 ± 189 pmol/min/mg. Western-blotting showed the the CYP3A4 protein expression in patients with chronic HBV infection was 3.04 ± 1.63, which was significantly lower than that of the normal control group ： 5.67 ± 1.52 （ P = 0. 003 ）. In contrast, no significant alteration of the Km of CYP3A4 was observed between two groups： 0. 142 ±0. 057μmol/L;0. 121 ±0. 024μmol/L （ P = 0. 103 ）. The enzymatic activity and protein expression of each liver sample had a high correlation. Correlation index was 0. 7683 （ P 〈 0. 001 ）. Conclusion Chronic HBV infection tends to down-regulate the expression of hepatic CYP3A4 and reduce its activity, but does not affect its structure. When patients with chronic HBV infection are given drugs metabolized by CYP3A4, the drugs＇side-effects brought by the variation of their metabolism should be taken into account.
出处
《中华医学杂志》
CAS
CSCD
北大核心
2006年第38期2703-2706,共4页
National Medical Journal of China
基金
国家自然科学基金(30571660)
