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该文研究决明子水提液对大鼠肝脏CYP450酶酶活性、mRNA及蛋白表达水平的影响。取SD大鼠,口服灌胃受试药物后,处死,用冰冷生理盐水灌流肝脏,提取大鼠肝脏微粒体、肝脏总RNA和总蛋白,采用Cocktail体外孵育法结合液质联用(LCMS)技术对大鼠肝脏中CYP1A2,2B1,2C11,2D2,2E1,3A1各亚酶的酶活性进行测定,并应用荧光定量PCR技术和Western blot对上述亚型的mRNA和蛋白表达水平进行检测。结果表明,酶活性方面,决明子水提液组与空白组比较可明显诱导CYP1A2,2B1,2C11,2D2,2E1,3A1的酶活性,其中决明子低剂量组对CYP2D2有明显的抑制作用,中、高剂量组对CYP2D2有明显的诱导作用,且都呈剂量依赖性增加,但对CYP3A1的诱导不呈剂量依赖性;mRNA表达方面,决明子水提液对CYP1A2,2C11,2D2,2E1mRNA的表达具有显著诱导作用,其中对CYP1A2,2D2,2E1诱导作用呈剂量依赖性,与酶活性水平具有一致性,对CYP2B1,3A1没有明显作用;蛋白表达方面,决明子水提液对CYP2C11,2E1的表达也具有诱导作用,但没有统计学差异。决明子水提液对CYP450同工酶不同程度诱导或抑制作用,特别是对于CYP2C11,2E1亚型酶,酶活性与mRNA,蛋白表达水平具有一致性,提示当与这些酶底物合并用药时,尤其是与CYP2C11,2E1代谢有关的药物合用时,应充分考虑到潜在的有益和不利的药物相互作用。
This paper studied the effect of aqueous extract of Cassia seed on rat liver CYP450 enzyme activity, mRNA and protein expression levels. Sprague-Dawley rats were used to orally ingest the test drug, then sacrificed. The liver was perfused with ice-cold physiological saline to extract rat liver microsomes, liver total RNA and total protein. Cocktail in vitro incubation and LCMS The enzyme activities of CYP1A2, 2B1, 2C11, 2D2, 2E1 and 3A1 were assayed in rat liver. The mRNA and protein levels of these subtypes were detected by fluorescence quantitative PCR and Western blot. The results showed that the activity of CYP1A2, 2B1, 2C11, 2D2, 2E1 and 3A1 was significantly induced by water extract of Cassia seed group compared with the blank group. The low dose Cassia seed group had a significant inhibitory effect on CYP2D2, The dose group had significant induction of CYP2D2 in a dose-dependent manner, but did not induce CYP3A1 in a dose-dependent manner. In mRNA expression, the aqueous extract of Cassia seed significantly induced the expression of CYP1A2, 2C11, 2D2 and 2E1 mRNA CYP1A2, 2D2 and 2E1 in a dose-dependent manner, which was consistent with the activity of CYP1A2, 2D2 and 2E1, but had no obvious effect on CYP2B1 and 3A1. In terms of protein expression, Cassia seed aqueous extract also induced the expression of CYP2C11 and 2E1 , But no statistical difference. The aqueous extract of Cassia to CYP450 isoenzymes induced or inhibited to varying degrees, especially for CYP2C11, 2E1 subtype enzymes, enzyme activity and mRNA, protein expression levels are consistent, suggesting that when combined with these enzyme substrates, especially Is CYP2C11, 2E1 metabolism associated with the drug should take full account of potentially beneficial and adverse drug interactions.