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目的研究丹红注射液对Ⅱ相药物代谢酶尿苷二磷酸葡萄糖醛酸转移酶(UGTs)的体外抑制作用。方法随机将新鲜的人原代肝细胞分为3组:实验组(丹红注射液)、阳性对照组(酮康唑)和空白对照组。分别将7个不同含量的丹红注射液(0.14%,0.41%,1.23%,3.70%,11.10%,33.30%和100.00%)、不同浓度的酮康唑溶液(1.37,4.12,12.30,37.00,111.00,333.00和1000.00μmol·L~(-1))10μL和纯化水与探针底物7-羟基香豆素(1000μmol·L~(-1))10μL置于人原代肝细胞体外孵育体系80μL中,共同孵育15 min。用液相色谱-串联质谱法测定代谢产物7-羟基香豆素-D-葡萄糖醛酸苷的含量(代表UGTs活性),并计算其IC50值(表示丹红注射液对UGTs的抑制程度)。结果含量分别为0.00%,0.01%,0.04%,0.12%,0.37%,1.11%,3.33%和10.00%的丹红注射液(在孵育体系中稀释了10倍)对UGTs的抑制率分别为0,-1.6%,3.2%,8.2%,5.2%,17.1%,38.3%和71.4%,且呈浓度依赖性抑制作用。与空白对照组比较,1.11%,3.33%,10.00%的丹红注射液差异均有统计学意义(P<0.05)。在人原代肝细胞体外孵育体系中,丹红注射液对UGTs的IC50值为4.9%。结论丹红注射液对人原代肝细胞UGTs有较弱的抑制作用,不太可能导致与UGTs相关的药物相互作用。
Objective To study the inhibitory effect of Danhong injection on the phase Ⅱ drug-metabolizing enzymes uridine diphosphate glucuronosyltransferase (UGTs) in vitro. Methods Fresh human primary hepatocytes were randomly divided into three groups: experimental group (Danhong injection), positive control group (ketoconazole) and blank control group. Seven different concentrations of Danhong injection (0.14%, 0.41%, 1.23%, 3.70%, 11.10%, 33.30% and 100.00%), different concentrations of ketoconazole solution (1.37, 4.12, 12.30, 37.00, 10μL of purified water and 10μL of probe substrate 7-hydroxycoumarin (1000μmol·L -1) were placed in an in vitro incubation system of human primary hepatocytes 80μL, co-incubated for 15 min. The metabolite 7-hydroxycoumarin-D-glucuronide (representing the activity of UGTs) was determined by liquid chromatography-tandem mass spectrometry and the IC50 value was calculated. This indicates the inhibitory effect of Danhong injection on UGTs. Results The inhibitory rates of Danhong injection (diluted 10 times in the incubation system) to UGTs were 0.00%, 0.01%, 0.04%, 0.12%, 0.37%, 1.11%, 3.33% and 10.00% -1.6%, 3.2%, 8.2%, 5.2%, 17.1%, 38.3% and 71.4%, respectively, in a concentration-dependent manner. Compared with the blank control group, the differences of Danhong injection of 1.11%, 3.33% and 10.00% were statistically significant (P <0.05). In human primary hepatocyte in vitro incubation system, Danhong injection of UGTs IC50 value of 4.9%. Conclusion Danhong injection has a weaker inhibitory effect on UGTs of human primary hepatocytes, which is unlikely to result in the drug interaction with UGTs.