中华耳鼻咽喉头颈外科杂志
中華耳鼻嚥喉頭頸外科雜誌
중화이비인후두경외과잡지
CHINESE JOURNAL OF OTORHINOLARYNGOLOGY HEAD AND NECK SURGERY
2010年
6期
491-496
,共6页
肖红俊%杨琛%何园园%郑娜
肖紅俊%楊琛%何園園%鄭娜
초홍준%양침%하완완%정나
喹啉酸%神经毒索类%螺旋神经节%细胞%培养的
喹啉痠%神經毒索類%螺鏇神經節%細胞%培養的
규람산%신경독색류%라선신경절%세포%배양적
Quinolinic acid%Neurotoxiins%Spiral ganglion%Cells,cultured
目的 探讨喹啉酸(quinolinic acid,QA)对大鼠螺旋神经节细胞(spiral ganglion cell,SGC)的神经兴奋毒性作用,观察N-甲基-D-天冬氨酸(N-methyl-D-aspartate,NMDA)受体拮抗剂5-甲基二氢丙环庚烯业胺马来酸(MK-801)对QA神经兴奋毒性的拮抗作用,同时观察镁离子对SGC的保护作用.方法 新生SD大鼠SGC原代培养72 h后,更换培养基,分为空白对照组、QA损伤组(1 mmol/L QA)、MK-801拮抗组(1 mmol/L QA+20 μmol/L MK-801)、MgCl2保护组(1 mmoL/L QA+1 mmol/L MgCl2),继续培养24 h后,通过磷脂酰丝氨酸外翻分析法在荧光显微镜下测定SGC凋亡率.SGC原代培养72 h后,分为四组:低浓度QA组(100 μmol/L QA),高浓度QA组(1 mmnol/LQA),MK-801拮抗组(20 μmol/L MK-801+l mmol/QA),MgCl2保护组(1 mmol/L MgCl2+l mmol/L QA);激光共聚焦显微镜动态检测各组SGC胞内瞬时钙离子浓度的变化.结果 与空白对照组(凋亡率9.2%±0.9%,(x)±s,下同)相比,QA损伤组可见大量SGC凋亡(凋亡率59.1%±7.5%),差异具有统计学意义(q=11.9,P<0.05);MgCl2保护组凋亡细胞比QA组明显减少,凋亡率为27.5%±8.3%,二者筹异具有统计学意义(q=7.5,P<0.05);MK-801组细胞凋亡率(12.8%±5.7%)与空白对照组差异无统计学意义(q=0.9,P>0.05).在高浓度QA(1 mmoL/L)的作用下,SGC内钙离子浓度明显升高,190 s时达高峰,随后逐渐下降,450 s时基本恢复加药前水平;低浓度QA(100 μmol/L)对SGC胞内钙离子浓度没有明赤影响;Mgcl2组SGC胞内钙离子浓度曲线峰值降低,时程缩短;MK-801组未见明显SGC胞内钙离子浓度变化.结论 QA可过度激活细胞膜上的NMDA受体而造成大鼠离体培养SGC的损伤,镁离子可以降低QA对SGC的神经兴奋毒性作用.
目的 探討喹啉痠(quinolinic acid,QA)對大鼠螺鏇神經節細胞(spiral ganglion cell,SGC)的神經興奮毒性作用,觀察N-甲基-D-天鼕氨痠(N-methyl-D-aspartate,NMDA)受體拮抗劑5-甲基二氫丙環庚烯業胺馬來痠(MK-801)對QA神經興奮毒性的拮抗作用,同時觀察鎂離子對SGC的保護作用.方法 新生SD大鼠SGC原代培養72 h後,更換培養基,分為空白對照組、QA損傷組(1 mmol/L QA)、MK-801拮抗組(1 mmol/L QA+20 μmol/L MK-801)、MgCl2保護組(1 mmoL/L QA+1 mmol/L MgCl2),繼續培養24 h後,通過燐脂酰絲氨痠外翻分析法在熒光顯微鏡下測定SGC凋亡率.SGC原代培養72 h後,分為四組:低濃度QA組(100 μmol/L QA),高濃度QA組(1 mmnol/LQA),MK-801拮抗組(20 μmol/L MK-801+l mmol/QA),MgCl2保護組(1 mmol/L MgCl2+l mmol/L QA);激光共聚焦顯微鏡動態檢測各組SGC胞內瞬時鈣離子濃度的變化.結果 與空白對照組(凋亡率9.2%±0.9%,(x)±s,下同)相比,QA損傷組可見大量SGC凋亡(凋亡率59.1%±7.5%),差異具有統計學意義(q=11.9,P<0.05);MgCl2保護組凋亡細胞比QA組明顯減少,凋亡率為27.5%±8.3%,二者籌異具有統計學意義(q=7.5,P<0.05);MK-801組細胞凋亡率(12.8%±5.7%)與空白對照組差異無統計學意義(q=0.9,P>0.05).在高濃度QA(1 mmoL/L)的作用下,SGC內鈣離子濃度明顯升高,190 s時達高峰,隨後逐漸下降,450 s時基本恢複加藥前水平;低濃度QA(100 μmol/L)對SGC胞內鈣離子濃度沒有明赤影響;Mgcl2組SGC胞內鈣離子濃度麯線峰值降低,時程縮短;MK-801組未見明顯SGC胞內鈣離子濃度變化.結論 QA可過度激活細胞膜上的NMDA受體而造成大鼠離體培養SGC的損傷,鎂離子可以降低QA對SGC的神經興奮毒性作用.
목적 탐토규람산(quinolinic acid,QA)대대서라선신경절세포(spiral ganglion cell,SGC)적신경흥강독성작용,관찰N-갑기-D-천동안산(N-methyl-D-aspartate,NMDA)수체길항제5-갑기이경병배경희업알마래산(MK-801)대QA신경흥강독성적길항작용,동시관찰미리자대SGC적보호작용.방법 신생SD대서SGC원대배양72 h후,경환배양기,분위공백대조조、QA손상조(1 mmol/L QA)、MK-801길항조(1 mmol/L QA+20 μmol/L MK-801)、MgCl2보호조(1 mmoL/L QA+1 mmol/L MgCl2),계속배양24 h후,통과린지선사안산외번분석법재형광현미경하측정SGC조망솔.SGC원대배양72 h후,분위사조:저농도QA조(100 μmol/L QA),고농도QA조(1 mmnol/LQA),MK-801길항조(20 μmol/L MK-801+l mmol/QA),MgCl2보호조(1 mmol/L MgCl2+l mmol/L QA);격광공취초현미경동태검측각조SGC포내순시개리자농도적변화.결과 여공백대조조(조망솔9.2%±0.9%,(x)±s,하동)상비,QA손상조가견대량SGC조망(조망솔59.1%±7.5%),차이구유통계학의의(q=11.9,P<0.05);MgCl2보호조조망세포비QA조명현감소,조망솔위27.5%±8.3%,이자주이구유통계학의의(q=7.5,P<0.05);MK-801조세포조망솔(12.8%±5.7%)여공백대조조차이무통계학의의(q=0.9,P>0.05).재고농도QA(1 mmoL/L)적작용하,SGC내개리자농도명현승고,190 s시체고봉,수후축점하강,450 s시기본회복가약전수평;저농도QA(100 μmol/L)대SGC포내개리자농도몰유명적영향;Mgcl2조SGC포내개리자농도곡선봉치강저,시정축단;MK-801조미견명현SGC포내개리자농도변화.결론 QA가과도격활세포막상적NMDA수체이조성대서리체배양SGC적손상,미리자가이강저QA대SGC적신경흥강독성작용.
Objective To investigate the neurotoxicity and its mechanism of quinolinic acid (QA) to spiral ganglion cells(SGC) and observe the protectable potential of MgCl2 on SGC. Methods SGC were cultured in vitro for 72 h, and then were divided into 4 groups; control group, QA group( 1 mmol/L QA) , MK-801 group (1 mmol/L QA + 20 μmol/L MK-801 ) and MgCl2 protected group ( 1 mmol/L QA + 1 mmol/L MgCl2). SGC apoptosis rate was analyzed by Annexin V staining and PI staining measurements after 24 h exposure to different medium. SGC cultured as methods above were divided into 4 groups as following: 100 (μmol/L QA, 1 mmol/L QA, 20 μmol/L MK-801 + 1 mmol/L QA and 1 mmol/L MgCl2 + 1 mmol/L QA. The intracellular calcium concentration was measured by laser scanning confocal microscope finally. Results Apoptosis rate in QA group was higher than that in both of control group (59. 1% ±7.5% vs 9.2% ±0. 9%, x±s, q = 11.9,P<0. 05) and MgCl2 group(59. 1% ±7.5% vs 27.5% ±8.3%, q =7.5 ,P <0.05). There was no significant difference between apoptosis rate of control and MK-801 group (12.8% ±5.7% vs 9.2% ±0.9% ,q = 0.9,P>0.05). It was shown that there was a significant increase of Ca2+ in SGC in the presence of QA by laser scanning confocal microscope. MK-801 may completely block the increase of Ca2+ , and the increase of Ca2+ can be reduce by the application of MgCl2. Conclusions QA might injure SGC by excessive activating NMDA receptors on the cell membrane. Mg2+ may have the function to reduce the neurotoxicity of QA.
