中国医药导报
中國醫藥導報
중국의약도보
CHINA MEDICAL HERALD
2014年
36期
4-8
,共5页
王彧杰%王媛媛%王蓉%原永芳
王彧傑%王媛媛%王蓉%原永芳
왕욱걸%왕원원%왕용%원영방
二甲亚砜%细胞色素P450酶3A4%细胞色素P450酶2C9%药物代谢%相互作用
二甲亞砜%細胞色素P450酶3A4%細胞色素P450酶2C9%藥物代謝%相互作用
이갑아풍%세포색소P450매3A4%세포색소P450매2C9%약물대사%상호작용
Dimethyl Sulfoxide%CYP3A4%CYP2C9%Drug metabolism%Drug interaction
目的：研究0.01%、0.05%、0.1%二甲亚砜(DMSO)对细胞色素P450(CYP450)酶系中3A4、2C9两个亚型基因及蛋白表达水平的影响。方法 Chang肝脏细胞经处理后，分为空白对照组(仅含培养液)、DMSO组(分别采用0.01%、0.05%、0.1%DMSO处理)、睾酮组(分别采用1、10、100μmol/L睾酮处理)、利福平组(分别采用1、10、100μmol/L利福平处理)、DMSO+睾酮组(分别采用0.01%、0.05%、0.1%DMSO+10μmol/L睾酮处理)、DMSO+利福平组(分别采用0.01%、0.05%、0.1%DMSO+10μmol/L利福平处理)。利用逆转录定量PCR的方法，测定CYP3A4、CYP2C9基因水平的表达。利用蛋白免疫印迹法，测定CYP3A4、CYP2C9蛋白水平的表达。结果0.1%DMSO组CYP3A4 mRNA高于空白对照组(P<0.01)，且0.1%DMSO组CYP3A4蛋白的表达也高于空白对照组。0.01%、0.05%DMSO组CYP3A4 mRNA表达与空白对照组比较差异无统计学意义(P>0.05)，但0.01%、0.05%DMSO组CYP3A4蛋白的表达高于空白对照组。0.01%、0.05%、0.1%DMSO组CYP2C9 mRNA表达与空白对照组比较差异无统计学意义(P>0.05)，0.01%、0.05%、0.1%DMSO组CYP2C9蛋白表达与空白对照组比较差异无统计学意义(P>0.05)。0.01%、0.05%DMSO+10μmol/L睾酮组CYP3A4 mRNA的表达与空白对照组比较差异无统计学意义(P>0.05)，但0.01%、0.05%DMSO+10μmol/L睾酮组CYP3A4蛋白的表达高于睾酮组。0.01%、0.05%、0.1%DMSO+10μmol/L利福平组CYP2C9 mRNA的表达与利福平组比较差异无统计学意义(P>0.05)，但0.01%、0.05%、0.1%DMSO+10μmol/L利福平组CYP2C9蛋白的表达高于利福平组。结论0.01%、0.05%、0.1%三个浓度的DMSO在体外肝细胞实验中对CYP3A4的表达有一定的影响，而对CYP2C9的表达影响不明显，当DMSO作为药物溶剂的时候，可能会影响药物单用时的实验结果。
目的：研究0.01%、0.05%、0.1%二甲亞砜(DMSO)對細胞色素P450(CYP450)酶繫中3A4、2C9兩箇亞型基因及蛋白錶達水平的影響。方法 Chang肝髒細胞經處理後，分為空白對照組(僅含培養液)、DMSO組(分彆採用0.01%、0.05%、0.1%DMSO處理)、睪酮組(分彆採用1、10、100μmol/L睪酮處理)、利福平組(分彆採用1、10、100μmol/L利福平處理)、DMSO+睪酮組(分彆採用0.01%、0.05%、0.1%DMSO+10μmol/L睪酮處理)、DMSO+利福平組(分彆採用0.01%、0.05%、0.1%DMSO+10μmol/L利福平處理)。利用逆轉錄定量PCR的方法，測定CYP3A4、CYP2C9基因水平的錶達。利用蛋白免疫印跡法，測定CYP3A4、CYP2C9蛋白水平的錶達。結果0.1%DMSO組CYP3A4 mRNA高于空白對照組(P<0.01)，且0.1%DMSO組CYP3A4蛋白的錶達也高于空白對照組。0.01%、0.05%DMSO組CYP3A4 mRNA錶達與空白對照組比較差異無統計學意義(P>0.05)，但0.01%、0.05%DMSO組CYP3A4蛋白的錶達高于空白對照組。0.01%、0.05%、0.1%DMSO組CYP2C9 mRNA錶達與空白對照組比較差異無統計學意義(P>0.05)，0.01%、0.05%、0.1%DMSO組CYP2C9蛋白錶達與空白對照組比較差異無統計學意義(P>0.05)。0.01%、0.05%DMSO+10μmol/L睪酮組CYP3A4 mRNA的錶達與空白對照組比較差異無統計學意義(P>0.05)，但0.01%、0.05%DMSO+10μmol/L睪酮組CYP3A4蛋白的錶達高于睪酮組。0.01%、0.05%、0.1%DMSO+10μmol/L利福平組CYP2C9 mRNA的錶達與利福平組比較差異無統計學意義(P>0.05)，但0.01%、0.05%、0.1%DMSO+10μmol/L利福平組CYP2C9蛋白的錶達高于利福平組。結論0.01%、0.05%、0.1%三箇濃度的DMSO在體外肝細胞實驗中對CYP3A4的錶達有一定的影響，而對CYP2C9的錶達影響不明顯，噹DMSO作為藥物溶劑的時候，可能會影響藥物單用時的實驗結果。
목적：연구0.01%、0.05%、0.1%이갑아풍(DMSO)대세포색소P450(CYP450)매계중3A4、2C9량개아형기인급단백표체수평적영향。방법 Chang간장세포경처리후，분위공백대조조(부함배양액)、DMSO조(분별채용0.01%、0.05%、0.1%DMSO처리)、고동조(분별채용1、10、100μmol/L고동처리)、리복평조(분별채용1、10、100μmol/L리복평처리)、DMSO+고동조(분별채용0.01%、0.05%、0.1%DMSO+10μmol/L고동처리)、DMSO+리복평조(분별채용0.01%、0.05%、0.1%DMSO+10μmol/L리복평처리)。이용역전록정량PCR적방법，측정CYP3A4、CYP2C9기인수평적표체。이용단백면역인적법，측정CYP3A4、CYP2C9단백수평적표체。결과0.1%DMSO조CYP3A4 mRNA고우공백대조조(P<0.01)，차0.1%DMSO조CYP3A4단백적표체야고우공백대조조。0.01%、0.05%DMSO조CYP3A4 mRNA표체여공백대조조비교차이무통계학의의(P>0.05)，단0.01%、0.05%DMSO조CYP3A4단백적표체고우공백대조조。0.01%、0.05%、0.1%DMSO조CYP2C9 mRNA표체여공백대조조비교차이무통계학의의(P>0.05)，0.01%、0.05%、0.1%DMSO조CYP2C9단백표체여공백대조조비교차이무통계학의의(P>0.05)。0.01%、0.05%DMSO+10μmol/L고동조CYP3A4 mRNA적표체여공백대조조비교차이무통계학의의(P>0.05)，단0.01%、0.05%DMSO+10μmol/L고동조CYP3A4단백적표체고우고동조。0.01%、0.05%、0.1%DMSO+10μmol/L리복평조CYP2C9 mRNA적표체여리복평조비교차이무통계학의의(P>0.05)，단0.01%、0.05%、0.1%DMSO+10μmol/L리복평조CYP2C9단백적표체고우리복평조。결론0.01%、0.05%、0.1%삼개농도적DMSO재체외간세포실험중대CYP3A4적표체유일정적영향，이대CYP2C9적표체영향불명현，당DMSO작위약물용제적시후，가능회영향약물단용시적실험결과。
Objective To investigate effects of 0.01%, 0.05%, 0.1% dimethyl sulfoxide (DMSO) on expression of CYP3A4 and CYP2C9 mRNA and protein. Methods Chang liver cells were divided into control group (dealed with RPMI-1640 singly), DMSO groups (dealed with 0.01%, 0.05%, 0.1%DMSO), testosterone groups (dealed with 1, 10, 100μmol/L testosterone), rifampicin groups (dealed with 1, 10, 100μmol/L Rifampicin), DMSO+testosterone groups (0.01%, 0.05%, 0.1%DMSO+10μmol/L testosterone) and DMSO+rifampicin groups (dealed with 0.01%, 0.05%, 0.1%DMSO+10μmol/L Rifampicin). The expression of CYP3A4 and CYP2C9 mRNA were detected by RT-qPCR, and the expression of CYP3A4 and CYP2C9 protein were detected by Western blot. Results Compared with control group, the expression of CYP3A4 mRNA and protein was increased by 0.1%DMSO (P<0.01);the expression of CYP3A4 protein could be in-duced by 0.01% and 0.05%DMSO, but no statistically significant difference on CYP3A4 mRNA (P>0.05). Compared with control group, there was no statistically significant difference on the expression of CYP2C9 mRNA and protein in DMSO groups (P> 0.05). Compared with control group, there was no statistically significant difference on the expres-sion of CYP3A4 mRNA in DMSO+testosterone groups (P>0.05). Compared with testosterone group, the expression of CYP3A4 protein was induced in 0.01%, 0.05%, 0.1%DMSO + testosterone group. Compared with rifampicin group, the expression of CYP2C9 protein was increased in 0.01%, 0.05%, 0.1% DMSO+Rifampicin group, but no statistically sig-nificant difference on CYP2C9 mRNA (P > 0.05). Conclusion The efficiency of 0.01%, 0.05% and 0.1% DMSO is greater on CYP3A4 than CYP2C9 in vitro test. 0.01%, 0.05%, 0.1% DMSO can influence expression of CYP3A4, but have poor effect on expression of CYP2C9 in vitro. DMSO as solvent can influence results of experiment.
