Mall M, Wissner A, Seydewitz H H, Hübner M, Kuehr J, Brandis M, Greger R, Kunzelmann K
Universitäts-Kinderklink, Albert-Ludwigs-Universität Freiburg, Mathildenstrabetae 1, 79106 Freiburg, Germany.
Br J Pharmacol. 2000 Aug;130(8):1884-92. doi: 10.1038/sj.bjp.0703520.
The flavonoid genistein has been shown to activate a Cl(-) conductance in various cell types expressing CFTR. We examined if similar effects can be observed when genistein is applied to native ex vivo tissues from human respiratory tract and rectum. We further compared the effects when genistein was applied to oocytes of Xenopus laevis expressing CFTR. In oocytes, both wtCFTR and DeltaF508-CFTR were activated by genistein while both cyclic AMP (K(v)LQT1) and Ca(2+) (SK4) activated K(+) channels were inhibited at high concentrations of genistein. Biopsies from nasal polyps and rectal mucosa were obtained from normal individuals (non-CF) and CF patients and in the presence of amiloride (10 micromol l(-1); mucosal side) the effects of genistein were assessed using a perfused Ussing chamber. In non-CF airway epithelia, genistein (50 micromol l(-1); mucosal side) increased lumen negative I(sc) but had no additional effects on tissues pre-stimulated with IBMX and forskolin (100 micromol l(-1) and 1 micromol l(-1); both sides). In non-CF rectal biopsies, in the presence of amiloride (10 micromol l(-1); mucosal side) and indomethacin (10 micromol l(-1); basolateral side), genistein increased lumen negative I(sc) and enabled cholinergic (carbachol; CCH, 100 micromol l(-1); basolateral side) stimulation of Cl(-) secretion indicating activation of luminal CFTR Cl(-) channels. However, after stimulation with IBMX/forskolin, genistein induced opposite effects and significantly inhibited CCH activated I(sc). In CF airway and intestinal tissues genistein failed to induce Cl(-) secretion. Thus, genistein is able to activate luminal CFTR Cl(-) conductance in non-CF tissues and mutant CFTR in oocytes. However, additional inhibitory effects on basolateral K(+) conductance and missing effects in native CF tissues do not support the use for pharmacological intervention in CF.
黄酮类化合物染料木黄酮已被证明可在多种表达囊性纤维化跨膜传导调节因子(CFTR)的细胞类型中激活氯离子(Cl⁻)电导。我们研究了将染料木黄酮应用于来自人类呼吸道和直肠的离体天然组织时是否能观察到类似的效果。我们还比较了将染料木黄酮应用于表达CFTR的非洲爪蟾卵母细胞时的效果。在卵母细胞中,野生型CFTR(wtCFTR)和ΔF508 - CFTR均被染料木黄酮激活,而在高浓度染料木黄酮作用下,环磷酸腺苷(K⁺通道亚基LQT1)激活的钾离子通道和钙离子(小电导钙激活钾通道4,SK4)激活的钾离子通道均受到抑制。从正常个体(非囊性纤维化,non - CF)和囊性纤维化(CF)患者获取鼻息肉和直肠黏膜活检组织,并在存在氨氯地平(10 μmol/L;黏膜侧)的情况下,使用灌注型尤斯灌流小室评估染料木黄酮的作用。在非CF气道上皮中,染料木黄酮(50 μmol/L;黏膜侧)增加了管腔负向短路电流(Isc),但对预先用异丁基甲基黄嘌呤(IBMX)和福斯可林(100 μmol/L和1 μmol/L;双侧)刺激的组织没有额外影响。在非CF直肠活检组织中,在存在氨氯地平(10 μmol/L;黏膜侧)和吲哚美辛(10 μmol/L;基底外侧)的情况下,染料木黄酮增加了管腔负向Isc,并使胆碱能(卡巴胆碱;CCH,100 μmol/L;基底外侧)刺激的Cl⁻分泌得以实现,表明管腔CFTR Cl⁻通道被激活。然而,在用IBMX/福斯可林刺激后,染料木黄酮产生了相反的效果,并显著抑制了CCH激活的Isc。在CF气道和肠道组织中,染料木黄酮未能诱导Cl⁻分泌。因此,染料木黄酮能够激活非CF组织中的管腔CFTR Cl⁻电导以及卵母细胞中的突变型CFTR。然而,对基底外侧钾离子电导的额外抑制作用以及在天然CF组织中缺乏效果并不支持将其用于CF的药物干预。