Puerarin Inhibits Acid-Sensing Ion Channels and Protects against Neuron Death Induced by Acidosis

Abstract

Puerarin, the main isoflavone found in the root of the wild leguminous creeper Pueraria lobata (Willd.) Ohwi, is known to protect the brain from ischemia-induced injuries. However, its effect on acidosis, a byproduct of brain ischemia and one factor promoting cell death, remains unknown. The purpose of this study was to investigate the effects of puerarin on acid-sensing ion channels (ASICs), which are reported to be activated by extracellular acidosis. Using the MTT assay and Hoechst/PI staining, we showed that puerarin could prevent hippocampal cell death during extracellular low pH. Data from whole-cell patch clamp recordings demonstrated that puerarin decreased the amplitude of ASIC currents and accelerated the desensitization of ASICs. Puerarin also reduced the amplitude of homomeric ASIC1a channels in CHO cells expressing pECFP‐ASIC1a. Collectively, our study demonstrated that puerarin prevented hippocampal cells from acidosis-induced death via ASIC blockage, providing a mechanical insight into the neuroprotective effects of puerarin during brain ischemia.

Key words

acid‐sensing ion channels - acidosis - puerarin - hippocampal neuron - Chinese hamster ovary cells

References

• 1 Siesjö B, Katsura K, Kristián T. Acidosis-related damage.  Adv Neurol. 1996;  71 209-236
• 2 Tombaugh G C S R. Evolving concepts about the role of acidosis in ischemic neuropathology.  J Neurochem. 1993;  61 793-803
• 3 Waldmann R, Champigny G, Bassilana F, Heurteaux C, Lazdunski M. A proton-gated cation channel involved in acid-sensing.  Nature. 1997;  386 173-177
• 4 de la Rosa D A, Krueger S R, Kolar A, Shao D, Fitzsimonds R M, Canessa C M. Distribution, subcellular localization and ontogeny of ASIC1 in the mammalian central nervous system.  J Physiol. 2003;  546 77-87
• 5 Chen C C, England S, Akopian A N, Wood J N. A sensory neuron-specific, proton-gated ion channel.  Proc Natl Acad Sci USA. 1998;  95 10240-10245
• 6 de la Rosa D A, Canessa C M, Fyfe G K, Zhang P. Structure and regulation of amiloride-sensitive sodium channels.  Ann Rev Physiol. 2000;  62 573-594
• 7 Wemmie J A, Chen J, Askwith C C, Hruska-Hageman A M, Price M P, Nolan B C, Yoder P G, Lamani E, Hoshi T, Freeman J H, Welsh M J. The acid-activated ion channel ASIC contributes to synaptic plasticity, learning, and memory.  Neuron. 2002;  34 463-477
• 8 Xiong Z G, Zhu X M, Chu X P, Minami M, Hey J, Wei W L, MacDonald J F, Wemmie J A, Price M P, Welsh M J, Simon R P. Neuroprotection in ischemia: blocking calcium-permeable acid-sensing ion channels.  Cell. 2004;  118 687-698
• 9 Gao J, Duan B, Wang D G, Deng X H, Zhang G Y, Xu L, Xu T L. Coupling between NMDA receptor and acid-sensing ion channel contributes to ischemic neuronal death.  Neuron. 2005;  48 635-646
• 10 Coryell M W, Ziemann A E, Westmoreland P J, Haenfler J M, Kurjakovic Z, Zha X M, Price M, Schnizler M K, Wemmie J A. Targeting ASIC1a reduces innate fear and alters neuronal activity in the fear circuit.  Biol Psychiatry. 2007;  62 1140-1148
• 11 Vukicevic M, Kellenberger S. Modulatory effects of acid-sensing ion channels on action potential generation in hippocampal neurons.  Am J Physiol. 2004;  287 682-690
• 12 Yermolaieva O, Leonard A S, Schnizler M K, Abboud F M, Welsh M J. Extracellular acidosis increases neuronal cell calcium by activating acid-sensing ion channel 1a.  Proc Natl Acad Sci USA. 2004;  101 6752-6757
• 13 Xiong Z G, Pignataro G, Li M, Chang S, Simon R P. Acid-sensing ion channels (ASICs) as pharmacological targets for neurodegenerative diseases.  Curr Opin Pharmacol. 2008;  8 25-32
• 14 Chen L, Chai Q, Zhao A, Chai X. Effect of puerarin on cerebral blood flow in dogs.  Zhongguo Zhong Yao Za Zhi. 1995;  20 560-562
• 15 Xu X, Zhang S, Zhang L, Yan W, Zheng X. The neuroprotection of puerarin against cerebral ischemia is associated with the prevention of apoptosis in rats.  Planta Med. 2005;  71 585-591
• 16 Tan Y, Liu M, Wu B. Puerarin for acute ischaemic stroke.  Cochrane Database Syst Rev. 2008;  (1) CD004955
• 17 Zhang G Q, Hao X M, Zhou P A, Wu C H, Dai D Z. Puerarin blocks transient outward K+ current and delayed rectifier K+ current in mice hippocampal CA1 neurons.  Acta Pharmacol Sin. 2001;  22 253-256
• 18 Zhang L, Deng T, Sun Y, Liu K, Yang Y, Zheng X. Role for nitric oxide in permeability of hippocampal neuronal hemichannels during oxygen glucose deprivation.  J Neurosci Res. 2008;  86 2281-2291
• 19 Gao Y, Liu S S, Qiu S, Cheng W, Zheng J, Luo J H. Fluorescence resonance energy transfer analysis of subunit assembly of the ASIC channel.  Biochem Biophys Res Commun. 2007;  359 143-150
• 20 Yang Y, Fukui K, Koike T, Zheng X X. Induction of autophagy in neurite degeneration of mouse superior cervical ganglion neurons.  Eur J Neurosci. 2007;  26 2979-2988
• 21 Nedergaard M, Goldman S A, Desai S, Pulsinelli W A. Acid-induced death in neurons and glia.  J Neurosci. 1991;  11 2489-2497
• 22 Xu X, Zheng X, Zhou Q, Li H. Inhibition of excitatory amino acid efflux contributes to protective effects of puerarin against cerebral ischemia in rats.  Biomed Environ Sci. 2007;  20 336-342
• 23 Nedergaard M, Kraig R P, Tanabe J, Pulsinelli W A. Dynamics of interstitial and intracellular pH in evolving brain infarct.  Am J Physiol Regul Integr Comp Physiol. 1991;  260 581-588
• 24 Cadiou H, Studer M, Jones N G, Smith E S J, Ballard A, McMahon S B, McNaughton P A. Modulation of acid-sensing ion channel activity by nitric oxide.  J Neurosci. 2007;  27 13251-13260
• 25 Xu X H, Zheng X X. Potential involvement of calcium and nitric oxide in protective effects of puerarin on oxygen–glucose deprivation in cultured hippocampal neurons.  J Ethnopharmacol. 2007;  113 421-426
• 26 Pignataro G, Simon R P, Xiong Z G. Prolonged activation of ASIC1a and the time window for neuroprotection in cerebral ischemia.  Brain. 2007;  130 151-158
• 27 Bano D, Young K, Guerin C, Lefeuvre R, Rothwell N, Naldini L, Rizzuto R, Carafoli E, Nicotera P. Cleavage of the plasma membrane Na⁺/Ca²⁺ exchanger in excitotoxicity.  Cell. 2005;  120 275-285