摘要
目的:研究辛芍组方中效应成分灯盏乙素、灯盏甲素和芍药苷在正常大鼠和大脑中动脉闭塞(MCAO)局灶性脑缺血再灌注损伤大鼠肝微粒体中的酶促反应动力学,比较上述成分在不同状态大鼠肝微粒体中的体外酶代谢动力学差异。方法:制备不同状态大鼠的肝微粒体,将辛芍组方与大鼠肝微粒体进行孵育,选择UPLC-MS为分析手段,采用底物消除法,计算各成分在正常大鼠和MCAO大鼠肝微粒体中的酶反应动力学米氏常数(Km),最大反应速率(Vmax)及体外肝微粒体对药物的固有清除率(CLint),将各参数进行组间统计学分析。结果:灯盏乙素、灯盏甲素和芍药苷在正常大鼠体外肝微粒体中的Km[(0. 597±0. 065),(0. 166±0. 012),(0. 640±0. 046)μmol·L~(-1)]与MCAO大鼠中的Km[(0. 798±0. 031),(0. 213±0. 017),(0. 499±0. 029)μmol·L-1]均明显不同;与正常组相比,MCAO大鼠体内的灯盏乙素和芍药苷的Vmax和CLint均明显减少(P <0. 05,P <0. 01),灯盏甲素的Vmax也显著降低(P <0. 05)。结论:辛芍组方的效应成分在脑缺血再灌注损伤大鼠肝微粒体中的代谢速率降低、消除减慢。
Objective: To investigate and compare enzymatic kinetics of scutellarin,apigenin-7-Oglucronide and paeoniflorin from Xinshao fomula in liver microsomes of sham-operated rats and middle cerebral artery occlusion(MCAO) rats with focal cerebral ischemia-reperfusion injury. Method: Xinshao fomula were incubated respectively with liver microsomes of sham-operated rats and MCAO rats, UPLC-MS and substrate elimination method was employed,Michaelis constant(Km),maximum velocity of enzymatic reaction(Vmax) and intrinsic clearance(CLint) of these three components from Xinshao fomula in liver microsomes of sham-operated rats and MCAO rats were calculated, these parameters between different groups were evaluated by statistical analysis. Result: The Kmvalues of scutellarin,apigenin-7-O-glucronide and paeoniflorin in liver microsomes of MCAO rats were(0. 798 ± 0. 031),(0. 213 ± 0. 017),(0. 499 ± 0. 029) μmol·L~(-1),which were quite different to these in liver microsomes of sham-operated rats. Compared with the sham-operated group,Vmaxand CLintvalues of scutellarin and paeoniflorin in liver microsomes of MCAO rats were significantly reduced(P < 0. 05, P <0. 01),and Vmaxof apigenin-7-O-glucronide in liver microsomes of MCAO rat was also significantly reduced(P <0. 05). Conclusion: Metabolic rates of these three active components from Xinshao fomula in liver microsomes of MCAO rats with focal cerebral ischemia-reperfusion injury decrease with low elimination rate.
引文
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