- 期刊
Lysophospholipids Attenuate Acetylcholine-evoked Ca(superscript 2+) Responses in Bovine Chromaffin Cells
水解磷酸脂抑制牛嗜鉻細胞被乙醯膽鹼刺激所引起的鈣離子反應
摘要
交感神經節前神經分泌乙醯膽鹼，刺激腎上腺嗜鉻細胞將兒茶酚胺釋放到血液中，以反應短期壓力。乙醯膽鹼活化尼古丁及蕈毒鹼受器，提高胞內鈣離子濃度([Ca(上标 2+)]i)因而釋放兒茶酚胺。而在血液中，由免疫細胞所分泌的水解磷酸脂(lysophospholipids, LPLs)，包括鞘胺醇1-磷酸鹽(sphingosine 1-phosphate, S1P)及溶血磷脂酸(lysophosphatidic acid, LPA)，則會引起許多不同的生理反應。為探討LPLs對嗜鉻細胞[Ca(上标 2+)]i變化的效應，細胞先以鈣離子指示劑Fura-2標定，以觀察[Ca(上标 2+)]i的變化。再以1μMS1P或LPA處理0、10、20、30、40、50、或60分鐘後，以乙醯膽鹼(100μM)、蕈毒鹼(100μM)、DMPP(10μM)、或高鉀溶液(50mM KCl)刺激5秒。結果顯示在LPA或S1P處理20分鐘後，乙醯膽鹼所引起的[Ca(上标 2+)]i上升，受到明顯的抑制。然而高鉀溶液所引起的[Ca(上标 2+)]i上升，並不會受到S1P前處理的影響，但會被LPA所增加。蕈毒鹼刺激所引起的反應，僅會受LPLs稍微抑制；然而以DMPP活化尼苦丁受器所引起的[Ca(上标 2+)]i反應，則會受到LPA及S1P的顯著抑制。這些結果顯示，LPLs可能主要是透過抑制尼古丁受器，而達到抑制乙醯膽鹼所引起的[Ca(上标 2+)]i上升。
並列摘要
Adrenal medulla chromaffin cells are stimulated by acetylcholine released from sympathetic preganglionic neurons and secret catecholamines into the blood to accommodate short-term stress. Acetylcholine activates nicotinic and muscarinic receptors to elevate the intracellular Ca(superscript 2+) concentration and causes the secretion of catecholamines. Lysophospholipids in the serum, including sphingosine 1-phosphate (S1P) and lysophosphatidic acid (LPA), are released primarily from immune cells during blood clotting and infection to modulate various physiological activities. To characterize the effects of lysophospholipids on Ca(superscript 2+) responses in bovine chromaffin cells, cells were loaded with the Ca(superscript 2+) indicator fura-2, and fluorescence intensity was used to monitor changes in intracellular Ca(superscript 2+) concentration. Cells treated with 1 μM S1P or LPA for 0, 10, 20, 30, 40, 50, or 60 min were stimulated with acetylcholine (100 μM), muscarine (100 μM), DMPP (10 μM), or high K(superscript +) buffer (50 mM KCl) for 5 sec. The results showed that the Ca(superscript 2+) responses evoked by acetylcholine were significantly inhibited after incubation in LPA or S1P for 20 min. The Ca(superscript 2+) response evoked by high K(superscript +) buffer was not inhibited by S1P pretreatment and was significantly facilitated by LPA pretreatment. Muscarine-evoked Ca(superscript 2+) responses were slightly attenuated by LPA and S1P pretreatments. Nicotine-evoked responses were inhibited in both LPA- and S1P-pretreated cells. Our findings demonstrate that nicotinic receptors may be the main targets of lysophospholipids in inhibiting acetylcholine-evoked Ca(superscript 2+) responses.