中国药理学通报
中國藥理學通報
중국약이학통보
CHINESE PHARMACOLOGICAL BULLETIN
2014年
7期
926-930,931
,共6页
周涛%王宇光%马增春%肖勇%汤响林%梁乾德%胡东华%肖成荣%谭洪玲%高月
週濤%王宇光%馬增春%肖勇%湯響林%樑乾德%鬍東華%肖成榮%譚洪玲%高月
주도%왕우광%마증춘%초용%탕향림%량건덕%호동화%초성영%담홍령%고월
银杏内酯B%利福平%细胞色素P450 3A4%孕烷X受体%荧光定量PCR技术%小干涉RNA
銀杏內酯B%利福平%細胞色素P450 3A4%孕烷X受體%熒光定量PCR技術%小榦涉RNA
은행내지B%리복평%세포색소P450 3A4%잉완X수체%형광정량PCR기술%소간섭RNA
Ginkgolide B%rifampicin%cytochrome P450 3A4%pregnane X receptor%quantitative PCR%small interfering RNA
目的：研究银杏内酯B（ginkgolide B）对CYP3A4的诱导作用，进一步验证是否通过激活孕烷 X 受体实现对CYP3A4 mRNA和蛋白水平的诱导表达。方法用不同浓度银杏内酯 B 处理 LS174T 细胞，通过 Q-PCR 法检测CYP3A4 mRNA表达变化，进一步利用本实验室构建的PXR-CYP3A4稳定转染HepG2工程细胞株，结合荧光素酶报告基因技术，显示检测银杏内酯B对PXR的转录激活活性的影响。蛋白质免疫印迹法检测CYP3 A4蛋白水平表达的变化；利用siRNA技术敲低PXR的mRNA表达，检测在PXR低表达的条件下银杏内酯B对CYP3A4 mRNA和蛋白水平表达影响。结果银杏内酯B可以浓度依赖性的使CYP3 A4基因及蛋白表达水平上调，报告基因结果显示，银杏内酯B能够浓度依赖性的增强PXR的转录激活活性，在PXR低表达的条件下，银杏内酯B对CYP3 A4诱导作用明显低于正常表达组PXR。结论以上表明银杏内酯B 通过激活PXR受体，促进其转录激活活性进而诱导CYP3 A4表达上调，同时对PXR自身的表达无明显影响。
目的：研究銀杏內酯B（ginkgolide B）對CYP3A4的誘導作用，進一步驗證是否通過激活孕烷 X 受體實現對CYP3A4 mRNA和蛋白水平的誘導錶達。方法用不同濃度銀杏內酯 B 處理 LS174T 細胞，通過 Q-PCR 法檢測CYP3A4 mRNA錶達變化，進一步利用本實驗室構建的PXR-CYP3A4穩定轉染HepG2工程細胞株，結閤熒光素酶報告基因技術，顯示檢測銀杏內酯B對PXR的轉錄激活活性的影響。蛋白質免疫印跡法檢測CYP3 A4蛋白水平錶達的變化；利用siRNA技術敲低PXR的mRNA錶達，檢測在PXR低錶達的條件下銀杏內酯B對CYP3A4 mRNA和蛋白水平錶達影響。結果銀杏內酯B可以濃度依賴性的使CYP3 A4基因及蛋白錶達水平上調，報告基因結果顯示，銀杏內酯B能夠濃度依賴性的增彊PXR的轉錄激活活性，在PXR低錶達的條件下，銀杏內酯B對CYP3 A4誘導作用明顯低于正常錶達組PXR。結論以上錶明銀杏內酯B 通過激活PXR受體，促進其轉錄激活活性進而誘導CYP3 A4錶達上調，同時對PXR自身的錶達無明顯影響。
목적：연구은행내지B（ginkgolide B）대CYP3A4적유도작용，진일보험증시부통과격활잉완 X 수체실현대CYP3A4 mRNA화단백수평적유도표체。방법용불동농도은행내지 B 처리 LS174T 세포，통과 Q-PCR 법검측CYP3A4 mRNA표체변화，진일보이용본실험실구건적PXR-CYP3A4은정전염HepG2공정세포주，결합형광소매보고기인기술，현시검측은행내지B대PXR적전록격활활성적영향。단백질면역인적법검측CYP3 A4단백수평표체적변화；이용siRNA기술고저PXR적mRNA표체，검측재PXR저표체적조건하은행내지B대CYP3A4 mRNA화단백수평표체영향。결과은행내지B가이농도의뢰성적사CYP3 A4기인급단백표체수평상조，보고기인결과현시，은행내지B능구농도의뢰성적증강PXR적전록격활활성，재PXR저표체적조건하，은행내지B대CYP3 A4유도작용명현저우정상표체조PXR。결론이상표명은행내지B 통과격활PXR수체，촉진기전록격활활성진이유도CYP3 A4표체상조，동시대PXR자신적표체무명현영향。
Aim To study the induction effect of Ginkgolide B on CYP3A4,and further verify the role of pregnane X receptor in CYP3 A4 induction expres-sion. Methods With different concentrations of Ginkgolide B treatment on LS174T cells,the CYP3A4 mRNA expression was detected by Q-PCR assay,fur-ther PXR-CYP3 A4 stable translation HepG2 cell lines were used to test the effect of Ginkgolide B on activity of PXR by reporter gene screening assay.CYP3A4 protein expression was detected by Western blot.PXR was knocked down with transfected with siRNA, CYP3 A4 mRNA and protein were detected in the con-dition of PXR low expression.Results The results revealed that the level of CYP 3 A 4 gene and protein expression were significantly increased by Ginkgolide B,and there was no induction effect on PXR.Reporter gene screening showed that Ginkgolide B could en-hance the transcriptional activity of PXR in a concen-tration-dependent manner.Under conditions of low ex-pression of PXR ,Ginkgolide B could also increase ex-pression of CYP3A4,but the induction folds were low-er than those of normal PXR group.Conclusion Ginkgolide B can signicantly up-regulate CYP3 A4 ex-pression via the PXR-CYP3 A4 pathway,and it has no effects on PXR gene expression.
