Clarke L L, Harline M C
Dalton Cardiovascular Research Center, University of Missouri-Columbia 65211, USA.
Am J Physiol. 1998 Apr;274(4):G718-26. doi: 10.1152/ajpgi.1998.274.4.G718.
The role of the cystic fibrosis transmembrane conductance regulator (CFTR) in cAMP-stimulated HCO3- secretion across the murine duodenum was investigated. Serosal-to-mucosal flux of HCO3- (Js-->m, in mu eq.cm-2.h-1) and short-circuit current (Isc; in mu eq.cm-2.h-1) were measured by the pH stat method in duodenum from CFTR knockout [CFTR(-)] and normal [CFTR(+)] mice. Under control conditions, forskolin increased Js-->m and Isc (+1.7 and +3.5, respectively) across the CFTR(+) but not CFTR(-) duodenum. Both the forskolin-stimulated delta Js-->m and delta Isc were abolished by the CFTR channel blocker 5-nitro-2-(3-phenylpropylamino)benzoate, whereas inhibition of luminal Cl-/HCO3- exchange by luminal Cl- removal or DIDS reduced the Js-->m by approximately 18% without a consistent effect on the delta Isc. Methazolamide also reduced the Js-->m by 39% but did not affect the delta Isc. When carbonic anhydrase-dependent HCO3- secretion was isolated by using a CO2-gassed, HCO3(-)-free Ringer bath, forskolin stimulated the Js-->m and Isc (+0.7 and +2.0, respectively) across CFTR(+) but not CFTR(-) duodenum. Under these conditions, luminal Cl- substitution or DIDS abolished the Js-->m but not the delta Isc. It was concluded that cAMP-stimulated HCO3- secretion across the duodenum involves 1) electrogenic secretion via a CFTR HCO3- conductance and 2) electroneutral secretion via a CFTR-dependent Cl-/HCO3- exchange process that is closely associated with the carbonic anhydrase activity of the epithelium.
研究了囊性纤维化跨膜传导调节因子(CFTR)在环磷酸腺苷(cAMP)刺激的小鼠十二指肠HCO₃⁻分泌中的作用。采用pH计法,在CFTR基因敲除[CFTR(-)]和正常[CFTR(+)]小鼠的十二指肠中测量HCO₃⁻从浆膜侧向黏膜侧的通量(Js→m,单位为μeq·cm⁻²·h⁻¹)和短路电流(Isc;单位为μeq·cm⁻²·h⁻¹)。在对照条件下,福斯可林增加了CFTR(+)十二指肠而非CFTR(-)十二指肠的Js→m和Isc(分别增加了1.7和3.5)。CFTR通道阻滞剂5-硝基-2-(3-苯丙基氨基)苯甲酸可消除福斯可林刺激的ΔJs→m和ΔIsc,而通过去除管腔Cl⁻或4,4'-二异硫氰酸根合芪-2,2'-二磺酸(DIDS)抑制管腔Cl⁻/HCO₃⁻交换可使Js→m降低约18%,但对ΔIsc无一致影响。甲酰唑胺也使Js→m降低了39%,但不影响ΔIsc。当使用通入CO₂且不含HCO₃⁻的林格氏液分离碳酸酐酶依赖性HCO₃⁻分泌时,福斯可林刺激了CFTR(+)十二指肠而非CFTR(-)十二指肠的Js→m和Isc(分别增加了0.7和2.0)。在这些条件下,管腔Cl⁻替代或DIDS消除了Js→m,但未消除ΔIsc。得出的结论是,cAMP刺激的十二指肠HCO₃⁻分泌涉及1)通过CFTR HCO₃⁻电导的电分泌,以及2)通过与上皮细胞碳酸酐酶活性密切相关的CFTR依赖性Cl⁻/HCO₃⁻交换过程的电中性分泌。