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Archive > Volume 10 Issue 3 > 2018,10(3):255-262 Prev Next

佛手柑內(nèi)酯在MDCK-MDR1細胞中所介導的調(diào)節(jié)長春新堿轉(zhuǎn)運的機制

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Mechanism underlying bergapten-mediated regulation of vincristine transport in MDCK-MDR1 cells

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    摘要:
    目的：研究白芷中佛手柑內(nèi)酯對長春新堿轉(zhuǎn)運的影響及其可能機制。方法用表觀通透性系數(shù)（Papp）測定長春新堿經(jīng)MDCK-MDR1細胞膜的轉(zhuǎn)運，以觀察佛手柑內(nèi)酯對長春新堿轉(zhuǎn)運的影響。采用分子對接預測佛手柑內(nèi)酯與P-糖蛋白（P-gp）的結(jié)合力。通過羅丹明123（Rho123）的蓄積和活性分析，確定了佛手柑內(nèi)酯對P-gp功能和P-gp ATP酶活性的影響。以1,6-二苯基-1,3- 5-己三烯（DPH）為研究對象，研究了佛手柑內(nèi)酯對膜流動性的影響，并用Western blot和實時定量pcr 方法：分析了佛手柑內(nèi)酯對P-gp蛋白和mRNA表達的影響。這些實驗闡明了佛手柑內(nèi)酯對長春新堿的轉(zhuǎn)運的影響，并允許探索佛手柑內(nèi)酯作用的可能機制。 結(jié)果：佛手柑內(nèi)酯能抑制長春新堿在MDCK-MDR1細胞中的轉(zhuǎn)運，而佛手柑內(nèi)酯和P-gp之間的結(jié)合力較弱。佛手柑內(nèi)酯能降低Rho123在MDCK-MDR1細胞中的蓄積，增加膜流動性，上調(diào)P-gp蛋白和mRNA的表達，但對P-gp ATP酶活性無影響。 結(jié)論：佛手柑內(nèi)酯抑制長春新堿轉(zhuǎn)運的可能機制與佛手柑內(nèi)酯介導的P-gp蛋白上調(diào)、mRNA表達、膜流動性和P-gp酶活性有關(guān)。

    Abstract:
    Objectives To investigate the effects of bergapten of Angelicae Dahuricae Radix on the transport of vincristine and its possible mechanism. Methods The apparent permeability coefficient (Papp) for the transport of vincristine through the membrane of MDCK-MDR1 cells was used as an indicator of the effect of bergapten on vincristine transport. Molecular docking was employed to predict the binding force between bergapten and P-glycoprotein (P-gp). The effects of bergapten on P-gp function and P-gp ATPase activity were determined by rhodamine 123 (Rho123) accumulation and activity analysis, respectively. 1,6-Diphenyl-1,3,5-hexatriene (DPH) was used to study the effects of bergapten on membrane fluidity, and Western blotting and quantitative real-time PCR assays were performed to analyze the effect of bergapten on the protein and mRNA expression of P-gp, respectively. These experiments clarified the effects of bergapten on the transport of vincristine and allowed exploration of the possible mechanism underlying the effects of bergapten. Results The results showed that bergapten could inhibit the transport of vincristine in MDCK-MDR1 cells, and the binding force between bergapten and P-gp was weaker. Bergapten could reduce the accumulation of Rh123 in MDCK-MDR1 cells, increase the membrane fluidity, and upregulate P-gp protein and mRNA expression but it had no effect on P-gp ATPase activity. Conclusions Overall, we concluded that the possible mechanism through which bergapten inhibits vincristine transport was related to the bergapten-mediated upregulation of P-gp protein and mRNA expression, membrane fluidity or P-gp enzyme activity.

    關(guān)鍵詞:
    佛手柑內(nèi)酯，長春新堿，轉(zhuǎn)運，MDCK-MDR1

    Keywords:


    Project Supported:
    The work was supported by a project of the National Natural Science Foundation of China (Grant No. 81303237), the Training Project of Young Scientists in Jiangxi Province (No. 20153BCB23019), China Postdoctoral Science Foundation (2016 M590606) and the National Natural Science Foundation of Jiangxi Province (20161ACB21020).

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