磁感应加热诱导汉防己甲素释放及对肿瘤相关双孔钾离子通道TASK-3的作用研究
|
摘要
目的:构建体外K_(2P) 9.1 (TASK-3)通道过度表达的非洲爪蟾卵母细胞模型;建立外部和磁感应加热2种加热方法,研究其对载汉防己甲素磁性纳米粒中药物释放行为的影响,进一步研究药物及载药磁性纳米粒对TASK-3通道电流的影响。方法:制备载汉防己甲素的聚乳酸-羟基乙酸共聚物(PLGA)磁性纳米粒,采用RP-HPLC考察外部和磁感应加热2种方法对药物释放的影响。采用双电极电压钳技术研究了汉防己甲素、Fe_3O_4-PLGA纳米粒和Tet-Fe_3O_4-PLGA纳米粒及运用磁感应加热诱导药物释放对卵母细胞表面TASK-3通道电流的影响。结果:药物释放量与速率呈现温度依赖性,汉防己甲素对Xenopus oocytes表面TASK-3通道电流有明显抑制作用并呈现剂量依赖性,未载药的PLGA磁性纳米粒对TASK3通道电流不产生影响,而Tet-Fe_3O_(4 )-PLGA NPs在2种加热体系下均显示出对TASK-3通道明显的抑制作用。结论:Tet-Fe_3O_4-PLGA NPs在磁感应作用下能可控性增加汉防己甲素对TASK3通道电流的阻滞作用,这种应激响应性药物递送系统有望成为基于双孔钾通道K_(2P)9.1靶向治疗的新方法,并有可能用于其他各种癌症治疗。
OBJECTIVE To construct a model of Xenopus laevis oocytes with overexpression of K_2P 9.1(TASK-3) channels in vitro, and to establish two heating methods, i.e. external and magnetic induction heating, thus to study their effects on drug release behavior in tetrandrine loaded magnetic nanoparticles, and to further study the effects of drugs and drug-loaded magnetic nanoparticles on TASK-3 channel currents. METHODS Poly(lactic-co-glycolic acid)(PLGA) magnetic nanoparticles loaded with tetrandrine were prepared. The effects of external and magnetic heating on drug release were investigated by RP-HPLC. The effects of tetrandrine, Fe_3O_4-PLGA NPs, and Tet-Fe_3O_4-PLGA NPs and induced drug release by magnetic induction heating on TASK-3 channel currents on the surface of oocytes were investigated with a two-electrode voltage-clamp technique. RESULTS The drug release and releaserate showed temperature dependence. Tetrandrine significantly inhibited TASK-3 channel currents on Xenopus oocytes in a dose-dependent manner. Unloaded PLGA magnetic nanoparticles had no effect on TASK3 channel currents, whereas Tet-Fe_3O_4-PLGA NPs showed significant inhibition of TASK-3 channels under both heating systems. CONCLUSION Tet-Fe_3O_4-PLGA NPs can increase the blocking effect of tetrandrine on TASK3 channel currents under magnetic induction. This stress-responsive drug delivery system has a great potential to be adopted as a new method for targeted therapy based on the two-pore potassium channel K_(2 P)9.1, and may be used for many other cancer treatments.
OBJECTIVE To construct a model of Xenopus laevis oocytes with overexpression of K_2P 9.1(TASK-3) channels in vitro, and to establish two heating methods, i.e. external and magnetic induction heating, thus to study their effects on drug release behavior in tetrandrine loaded magnetic nanoparticles, and to further study the effects of drugs and drug-loaded magnetic nanoparticles on TASK-3 channel currents. METHODS Poly(lactic-co-glycolic acid)(PLGA) magnetic nanoparticles loaded with tetrandrine were prepared. The effects of external and magnetic heating on drug release were investigated by RP-HPLC. The effects of tetrandrine, Fe_3O_4-PLGA NPs, and Tet-Fe_3O_4-PLGA NPs and induced drug release by magnetic induction heating on TASK-3 channel currents on the surface of oocytes were investigated with a two-electrode voltage-clamp technique. RESULTS The drug release and releaserate showed temperature dependence. Tetrandrine significantly inhibited TASK-3 channel currents on Xenopus oocytes in a dose-dependent manner. Unloaded PLGA magnetic nanoparticles had no effect on TASK3 channel currents, whereas Tet-Fe_3O_4-PLGA NPs showed significant inhibition of TASK-3 channels under both heating systems. CONCLUSION Tet-Fe_3O_4-PLGA NPs can increase the blocking effect of tetrandrine on TASK3 channel currents under magnetic induction. This stress-responsive drug delivery system has a great potential to be adopted as a new method for targeted therapy based on the two-pore potassium channel K_(2 P)9.1, and may be used for many other cancer treatments.
引文
[1] Kim Y, Bang H, Kim D. TASK-3, a new member of the tandem pore K(+) channel family [J]. J Biol Chem, 2000,275:9340-9347.
[2] Pei L, Wiser O, Slavin A, et al. Oncogenic potential of TASK3 (Kcnk9) depends on K+ channel function [J]. ProcNatlAcadSci USA, 2003,100:7803-7807.
[3] Williams S, Bateman A, O’Kelly I. Altered Expression of Two-Pore Domain Potassium (K(2P)) Channels in Cancer [J]. PLoS ONE,2013, 8:e74589.
[4] Huang X, Jan LY. Targeting potassium channels in cancer [J]. J Cell Bio,2014, 206:151-162.
[5] InnamaaAea. Expression and prognostic significance of the oncogenic K2P potassium channel KCNK9 (TASK-3) in ovarian carcinoma [J]. Anticancer Res, 2013,33:1401-1408.
[6] Gang Wang, Jose R. Lemos., Iadecola. C. Herbal alkaloid tetrandrine: from an ion channel blocker to inhibitor of tumor proliferation [J]. Trends Pharmacol Sci ,2004,25:120-3.
[7] Chen CS,Fu DH,Zheng SW,et al. Effect of inductive heating on drug release of tetrandrine-loaded PLGA magnetic nanoparticles [J].Chin Pharm J(中国药学杂志),2015,50(21):1893-1898.
[8] Makadia HK, Siegel SJ. Poly Lactic-co-Glycolic Acid (PLGA) as Biodegradable Controlled Drug Delivery Carrier [J]. Polymers,2011,3(3): 1377-1397.
[9] Friend DR. Factors affecting the degradation and drug-release mechanism of poly(lactic acid) and poly[(lactic acid)-co-(glycolic acid) [J].Polym Int, 2005,54 (1) :36-46.
[10] Externbrink A, Clark MR, Friend DR,et al. Investigating the feasibility of temperature-controlled accelerated drug release testing for an intravaginal ring [J]. Eur J Pharm Bio Pharm, 2013,85 (3): 966-973.
[11] Shameem M, Lee H, DeLuca PP.A short term (accelerated release) approach to evaluate peptide release from PLGA depot-formulations [J].AAPS Pharm Sci, 1999,1 (3): E7.
[12] Keles H, Naylor A, Clegg F, et al.Investigation of factors influencing the hydrolytic degradation of single PLGA microparticles [J]. Polym Degrad Stabil, 2015,119 :228-241.
[13] Ford Versypt AN, Pack DW, Braatz RD. Mathematical modeling of drug delivery from autocatalytically degradable PLGA microspheres-A review [J].J Controll Release, 2013,165 (1):29-37.
[14] Qi F, Wu J, Fan Q, et al. Preparation of uniform-sized exenatide-loaded PLGA microspheres as long-effective release system with high encapsulation efficiency and bio-stability [J]. Colloids Surf B Biointerfaces, 2013,112 : 492-498.
[15] Xu Q, Chin SE, Wang CH, et al. Mechanism of drug release from double-walled PDLLA(PLGA) microspheres [J]. Biomaterials, 2013,34 (15) :3902-3911.
[16] Corrigan OI, Li X.Quantifying drug release from PLGA nanoparticulates [J], Eur J Pharm Sci, 2009,37 (3-4): 477-485.
[17] Budhian A, Siegel SJ, Winey KI. Controlling the in vitro release profiles for a system of haloperidol-loaded PLGA nanoparticles [J]. Int J Pharm, 2008,346 (1-2): 151-159.
[18] Kang D, Choe C, Kim D. Thermosensitivity of the two-pore domain K+ channels TREK-2 and TRAAK [J]. J Physiol, 2005,564 (Pt1): 103-116.
[2] Pei L, Wiser O, Slavin A, et al. Oncogenic potential of TASK3 (Kcnk9) depends on K+ channel function [J]. ProcNatlAcadSci USA, 2003,100:7803-7807.
[3] Williams S, Bateman A, O’Kelly I. Altered Expression of Two-Pore Domain Potassium (K(2P)) Channels in Cancer [J]. PLoS ONE,2013, 8:e74589.
[4] Huang X, Jan LY. Targeting potassium channels in cancer [J]. J Cell Bio,2014, 206:151-162.
[5] InnamaaAea. Expression and prognostic significance of the oncogenic K2P potassium channel KCNK9 (TASK-3) in ovarian carcinoma [J]. Anticancer Res, 2013,33:1401-1408.
[6] Gang Wang, Jose R. Lemos., Iadecola. C. Herbal alkaloid tetrandrine: from an ion channel blocker to inhibitor of tumor proliferation [J]. Trends Pharmacol Sci ,2004,25:120-3.
[7] Chen CS,Fu DH,Zheng SW,et al. Effect of inductive heating on drug release of tetrandrine-loaded PLGA magnetic nanoparticles [J].Chin Pharm J(中国药学杂志),2015,50(21):1893-1898.
[8] Makadia HK, Siegel SJ. Poly Lactic-co-Glycolic Acid (PLGA) as Biodegradable Controlled Drug Delivery Carrier [J]. Polymers,2011,3(3): 1377-1397.
[9] Friend DR. Factors affecting the degradation and drug-release mechanism of poly(lactic acid) and poly[(lactic acid)-co-(glycolic acid) [J].Polym Int, 2005,54 (1) :36-46.
[10] Externbrink A, Clark MR, Friend DR,et al. Investigating the feasibility of temperature-controlled accelerated drug release testing for an intravaginal ring [J]. Eur J Pharm Bio Pharm, 2013,85 (3): 966-973.
[11] Shameem M, Lee H, DeLuca PP.A short term (accelerated release) approach to evaluate peptide release from PLGA depot-formulations [J].AAPS Pharm Sci, 1999,1 (3): E7.
[12] Keles H, Naylor A, Clegg F, et al.Investigation of factors influencing the hydrolytic degradation of single PLGA microparticles [J]. Polym Degrad Stabil, 2015,119 :228-241.
[13] Ford Versypt AN, Pack DW, Braatz RD. Mathematical modeling of drug delivery from autocatalytically degradable PLGA microspheres-A review [J].J Controll Release, 2013,165 (1):29-37.
[14] Qi F, Wu J, Fan Q, et al. Preparation of uniform-sized exenatide-loaded PLGA microspheres as long-effective release system with high encapsulation efficiency and bio-stability [J]. Colloids Surf B Biointerfaces, 2013,112 : 492-498.
[15] Xu Q, Chin SE, Wang CH, et al. Mechanism of drug release from double-walled PDLLA(PLGA) microspheres [J]. Biomaterials, 2013,34 (15) :3902-3911.
[16] Corrigan OI, Li X.Quantifying drug release from PLGA nanoparticulates [J], Eur J Pharm Sci, 2009,37 (3-4): 477-485.
[17] Budhian A, Siegel SJ, Winey KI. Controlling the in vitro release profiles for a system of haloperidol-loaded PLGA nanoparticles [J]. Int J Pharm, 2008,346 (1-2): 151-159.
[18] Kang D, Choe C, Kim D. Thermosensitivity of the two-pore domain K+ channels TREK-2 and TRAAK [J]. J Physiol, 2005,564 (Pt1): 103-116.