摘要
目的研究旱莲草5种中药单体木犀草素、芹菜素、槲皮素、刺囊酸和蟛蜞菊内酯是否可以通过PXR的激活而诱导CYP3A4转录表达。方法采用MTT法检测5种中药单体对人结肠癌细胞系LS174T细胞的毒性,采用脂质体瞬时转染的方法,在LS174T细胞中同时转入人PXR表达质粒、CYP3A4报告质粒以及内参质粒,建立双荧光报告基因体系,与不同浓度及不同给药时间的中药单体共同孵育,检测细胞中荧光素酶。结果 5种中药单体对LS174T的IC50值分别为木犀草素(71.08±5.35)μmol/L、芹菜素(185.54±9.57)μmol/L、槲皮素(112.68±8.34)μmol/L、蟛蜞菊内酯(36.01±9.37)μmol/L、刺囊酸(26.04±5.62)μmol/L。刺囊酸(5、10和20μmol/L)分别诱导PXR调控的CYP3A4表达(1.52±0.31)倍、(1.95±0.31)倍和(2.57±0.23)倍,蟛蜞菊内酯(5、10和20μmol/L)分别诱导PXR调控的CYP3A4表达(1.57±0.30)倍、(1.91±0.25)倍和(2.14±0.51)倍,木犀草素、槲皮素、芹菜素在试验的浓度范围内不能明显诱导LS174T细胞中PXR调控的CYP3A4表达。在不同给药时间试验中,10μmol/L和20μmol/L的刺囊酸在12～48 h能诱导PXR调控的CYP3A4表达,诱导能力随时间的延长而呈增强的趋势,各浓度的最大诱导倍数均在48 h出现;10μmol/L和20μmol/L的蟛蜞菊内酯只能在12～24 h诱导PXR调控的CYP3A4表达,在48 h时2个浓度组均出现诱导表达倍数的下调。结论刺囊酸和蟛蜞菊内酯在LS174T细胞系中可以通过激活PXR诱导CYP3A4转录表达,而木犀草素、芹菜素、槲皮素无此作用。
Objective To observe the effects of luteolin, apigenin, quercetin, wedelolactone and echinocystic acid, which are the monomers in Traditional Chinese Medicine Eclipta prostrata, on the human PXR mediated transcriptional regulation of CYP3A4. Methods The cytotoxic effect of the 5 Traditional Chinese Medicine monomers on LS174T were determined by MTY method, transient cotransfection reporter gene assays were performed on LS174T ceils with the PXR expression plasmid and the CYP3A4 reporter plasmid linked to luciferase. Results 50% inhibition concentrations （IC50） of luteolin, apigenin, quercetin, wedelolactone and echinocystic acid were （71.08 ±5.35）, （ 185.54 ±9.57）, （ 112.68±8.34 ）, （ 36.01 ± 9.37 ） and （26.04 ± 5.62 ） tzmol/L, respectively. Echinocystic acid and wedelolactone induced the CYP3A4 transcription through PXR, in an evident dose-dependent manner, but the CYP3A4 transcriptions in LSI74T ceils were not observed in luteolin, apigenin and quercetin. Different concentrations of echinocytic acid including 5, 10 and 20 ixmol/L exhibited a （ 1.52 ± 0. 31 ） fold, （ 1.95 ± 0. 31 ） fold and （2.57 ± 0.23 ） fold, respectively, compared with 0. 1% DMSO treated eells; different concentrations of wedelolactone including 5, 10 and 20 μmoL/L exhibited a （ 1.57 ±0.30） fold, （ 1.91 ±0.25） fold and （2.14 ±0.51 ） fold, respectively, compared with 0.1% DMSO treated ceils. In the time effect study, 10 and 20μmol/L of echinocystic acid inducecl CYP3/k4 transcription bet, seen 12 h and 48 h, the strongest induction appeared in 48 h, but 10 and 20 μmol/L of wedelolactone induced CYP3A4 transcription only in 12 h and 24 h, not in 48 h. Conclusion Echinocystic acid and wedelolactone may induce the CYP3A4 transcription through PXR, but in LS174T ceils luteolin, apigenin and quercetin have no effect on PXR-mediated transcription of CYP3A4.
出处
《今日药学》
CAS
2012年第3期129-133,141,共6页
Pharmacy Today
基金
国家自然科学基金(编号:30972917)
广州市科技局重大科技专项-药物制剂关键技术及产业化示范(编号:2008A1-E4101-7)
