酮洛芬异丙酯在HaCaT细胞中的代谢

 目的研究酮洛芬异丙酯在HaCaT细胞中的代谢作用，为探讨HaCaT细胞作为研究酯类前体药物的皮肤代谢模型提供实验依据。方法采用含不同浓度酮洛芬异丙酯的人角质形成细胞无血清培养基进行HaCaT细胞培养，或将不同量酮洛芬异丙酯加入HaCaT细胞的PBS匀浆中进行37℃温孵实验，通过高效液相色谱法分别在不同时间测定细胞培养液和细胞匀浆中酮洛芬异丙酯及酮洛芬的浓度。结果酮洛芬异丙酯在HaCaT细胞培养和匀浆中能被水解成酮洛芬。酮洛芬的形成速率与酮洛芬异丙酯初始浓度的大小有关，在低浓度时随酮洛芬异丙酯初始浓度增大而增加。细胞培养中，加入1.478 9，4.735 3，9.034 1，11.851 1μmol·L^-1的酮洛芬异丙酯对细胞活性影响不大，酮洛芬的形成速率分别是0.069 6，0.272 2，0.886 0，0.675 1μmol·L^-1·h^-1。细胞匀浆中，酮洛芬异丙酯的初始浓度是7.457 3，12.678 7，24.784 5，44.694 3 μmol·L^-1，相应的酮洛芬形成速率分别为0.134 7，0.278 2，0.473 5，0.608 9μmol·L^-1·h^-1。结论HaCaT细胞作为一种常用的角质形成细胞系细胞，可用于研究酯类前体药物的皮肤代谢。

Studies on metabolism of ketoprofen isopropyl ester in HaCaT cells

OBJECTIVE To investigate HaCaT cell as the model of ester prodrug metabolism in skin and study the metabolism of Ketoprofen isopropyl ester (KPE)in HaCaT cells.METHOD SThe human keratinocyte serum free medium (K SFM) with different concentrations of KPE was used to culture HaCaT cells, and the HaCaT cells homogenate in Phosphate Buffer Solution (PBS) with different concentrations of KPE was incubated at 37℃.The concentrations of KPE and Ketoprofen (KP) in different samples were measured using High Performance Liquid Chromatography (HPLC).RESULTS KPE was hydrolyzed into KP in HaCaT cells culture and HaCaT cells homogenate.KP formation rate was associated with the initial concentration of KPE.In HaCaT cell culture, the viability of HaCaT cell was not affected by KPE, and the KP formation rate was 0.0696，0.2722，0.8860 and 0.6751 μmol·L^-1·h^-1at 1.478 9，4.735 3，9.034 1 and 11.851 1 μmol·L^-1 KPE initial concentrations respectively.In cell homogenate, the KP formation rate respectively was 0.134 7，0.278 2，0.473 5 and 0.608 9 μmol·L^-1·h^-1 at 7.457 3，12.678 7，24.784 5 and 44.694 3 μmol·L^-1 KPE initial concentrations.CONCLUSION HaCaT cell as the cell of a frequently used human transformed keratinocyte cell line,can be used to study skin metabolism of ester prodrug.