中华老年医学杂志
中華老年醫學雜誌
중화노년의학잡지
Chinese Journal of Geriatrics
2010年
4期
319-323
,共5页
朱红灿%蔡春生%耿利娇%臧卫东%张华%任秀花
硃紅燦%蔡春生%耿利嬌%臧衛東%張華%任秀花
주홍찬%채춘생%경리교%장위동%장화%임수화
帕金森病%尿酸%运动神经元%氧化应激%膜电位,线粒体
帕金森病%尿痠%運動神經元%氧化應激%膜電位,線粒體
파금삼병%뇨산%운동신경원%양화응격%막전위,선립체
Parkinson disease%Uric acid%Motor neurons%Oxidative stress%Membrane potential,mitochondrial
目的 探讨尿酸对6-羟多巴胺(6-OHDA)诱导的SD大鼠帕金森病体外模型多巴胺(DA)能神经元氧化应激损伤的影响. 方法取孕12～14 d SD大鼠中脑原代细胞进行培养.实验分3组:(1)对照组:原代培养细胞;(2)6-OHDA组:原代培养细胞加6-OHDA;(3)尿酸组:不同浓度尿酸(5、50、100、250、500 μmol/L)分为5个亚组.培养第5天开始加尿酸干预,持续作用5 d,于第10天加50μmol/L的6-OHDA,作用2 h,培养第10天收集细胞.经酪氨酸羟化酶(TH)免疫细胞化学染色,3-(4,5-二甲基噻唑-2)-2,5-二苯基四氮唑溴盐(MTT)法检测细胞活力;通过流式细胞仪,用罗丹明123(Rh123)检测线粒体膜电位(△ψm). 结果对照组TH阳性(TH~+)细胞[(296.8±42.5)个/ml]较多,突起较长,部分密集成网状;6-OHDA组TH~+细胞[(92.8±19.7)个/ml]明显减少,突起较短,部分有断裂;5、50、100、250、500 μmol/L尿酸组TH~+细胞数(96.5±20.1、115.5± 30.0、152.5±26.7、205.0±48.2、230.1±22.5)个/ml,较对照组减少,但明显多于6-OHDA组,并且同尿酸浓度成正相关(F=13.94,P<0.05).与对照组比较,6-OHDA组细胞活力明显下降(F=90.19,P<0.05);5、50、100、500μmol/L尿酸组细胞活力较对照组下降(F=14.56,P<0.05),但高于6-OHDA组(F=40.96,P<0.05).250 μmol/L尿酸组细胞活力与对照组比较,差异无统计学意义(F=1.27,P>0.05).对照组△ψm(30.05±5.88)%,与6-OHDA组(23.67±2.72)比较明显下降(F=6.30,P<0.05);而尿酸各组与6-OHDA组比较,均升高;其中100、250μmol/L尿酸组△ψm(36.91±2.44)%、(38.08±2.90)%高于对照组(F=4.62,P<0.05). 结论尿酸可减少6-OHDA对神经元的毒性作用,提高细胞活力,稳定细胞膜电位,表明尿酸能通过抗氧化应激活性发挥其对多巴胺能神经元的保护作用.
目的 探討尿痠對6-羥多巴胺(6-OHDA)誘導的SD大鼠帕金森病體外模型多巴胺(DA)能神經元氧化應激損傷的影響. 方法取孕12～14 d SD大鼠中腦原代細胞進行培養.實驗分3組:(1)對照組:原代培養細胞;(2)6-OHDA組:原代培養細胞加6-OHDA;(3)尿痠組:不同濃度尿痠(5、50、100、250、500 μmol/L)分為5箇亞組.培養第5天開始加尿痠榦預,持續作用5 d,于第10天加50μmol/L的6-OHDA,作用2 h,培養第10天收集細胞.經酪氨痠羥化酶(TH)免疫細胞化學染色,3-(4,5-二甲基噻唑-2)-2,5-二苯基四氮唑溴鹽(MTT)法檢測細胞活力;通過流式細胞儀,用囉丹明123(Rh123)檢測線粒體膜電位(△ψm). 結果對照組TH暘性(TH~+)細胞[(296.8±42.5)箇/ml]較多,突起較長,部分密集成網狀;6-OHDA組TH~+細胞[(92.8±19.7)箇/ml]明顯減少,突起較短,部分有斷裂;5、50、100、250、500 μmol/L尿痠組TH~+細胞數(96.5±20.1、115.5± 30.0、152.5±26.7、205.0±48.2、230.1±22.5)箇/ml,較對照組減少,但明顯多于6-OHDA組,併且同尿痠濃度成正相關(F=13.94,P<0.05).與對照組比較,6-OHDA組細胞活力明顯下降(F=90.19,P<0.05);5、50、100、500μmol/L尿痠組細胞活力較對照組下降(F=14.56,P<0.05),但高于6-OHDA組(F=40.96,P<0.05).250 μmol/L尿痠組細胞活力與對照組比較,差異無統計學意義(F=1.27,P>0.05).對照組△ψm(30.05±5.88)%,與6-OHDA組(23.67±2.72)比較明顯下降(F=6.30,P<0.05);而尿痠各組與6-OHDA組比較,均升高;其中100、250μmol/L尿痠組△ψm(36.91±2.44)%、(38.08±2.90)%高于對照組(F=4.62,P<0.05). 結論尿痠可減少6-OHDA對神經元的毒性作用,提高細胞活力,穩定細胞膜電位,錶明尿痠能通過抗氧化應激活性髮揮其對多巴胺能神經元的保護作用.
목적 탐토뇨산대6-간다파알(6-OHDA)유도적SD대서파금삼병체외모형다파알(DA)능신경원양화응격손상적영향. 방법취잉12～14 d SD대서중뇌원대세포진행배양.실험분3조:(1)대조조:원대배양세포;(2)6-OHDA조:원대배양세포가6-OHDA;(3)뇨산조:불동농도뇨산(5、50、100、250、500 μmol/L)분위5개아조.배양제5천개시가뇨산간예,지속작용5 d,우제10천가50μmol/L적6-OHDA,작용2 h,배양제10천수집세포.경락안산간화매(TH)면역세포화학염색,3-(4,5-이갑기새서-2)-2,5-이분기사담서추염(MTT)법검측세포활력;통과류식세포의,용라단명123(Rh123)검측선립체막전위(△ψm). 결과대조조TH양성(TH~+)세포[(296.8±42.5)개/ml]교다,돌기교장,부분밀집성망상;6-OHDA조TH~+세포[(92.8±19.7)개/ml]명현감소,돌기교단,부분유단렬;5、50、100、250、500 μmol/L뇨산조TH~+세포수(96.5±20.1、115.5± 30.0、152.5±26.7、205.0±48.2、230.1±22.5)개/ml,교대조조감소,단명현다우6-OHDA조,병차동뇨산농도성정상관(F=13.94,P<0.05).여대조조비교,6-OHDA조세포활력명현하강(F=90.19,P<0.05);5、50、100、500μmol/L뇨산조세포활력교대조조하강(F=14.56,P<0.05),단고우6-OHDA조(F=40.96,P<0.05).250 μmol/L뇨산조세포활력여대조조비교,차이무통계학의의(F=1.27,P>0.05).대조조△ψm(30.05±5.88)%,여6-OHDA조(23.67±2.72)비교명현하강(F=6.30,P<0.05);이뇨산각조여6-OHDA조비교,균승고;기중100、250μmol/L뇨산조△ψm(36.91±2.44)%、(38.08±2.90)%고우대조조(F=4.62,P<0.05). 결론뇨산가감소6-OHDA대신경원적독성작용,제고세포활력,은정세포막전위,표명뇨산능통과항양화응격활성발휘기대다파알능신경원적보호작용.
Objective To study the effects of uric acid (UA) on oxidative stress of dopaminergic neurons of rat model of Parkinson's disease induced by 6-hydroxydopamine (6-OHDA). Methods Mesencephalic neurons in culture were prepared from embryonic Sprague-Dawley rat of 12-14 days. Various groups were categorized as follows: control group, 6-OHDA group and UA group which was sub divided into five subgroups according to different concentrations of UA (5, 50, 100, 250 and 500 μmol/L). Cultures were added with UA from the 5th day to the 10th day. At the 10th day, the cultures were co-treated with the toxin 6-OHDA (50 μmol/L) for 2 h, the cells were collected. The dopaminergic neurons were identified by tyrosine hydroxylase (TH) immunocytochemistry. The rate of cell viability was evaluated by MTT assay. By measuring the intracellular rhodamine 123 fluorescence density, mitochondrial membrane potential (△ψm) was evaluated. Results There was more TH-positive (TH ) neurons in the control group (296.8± 42.5), moreover, some cross-linked network structure could be found. As compared with the control group, there were less TH~+ neurons in 6-OHDA group (92.8±19. 7, F=75.26, P<0.05). More TH~+ neurons were found in 5, 50, 100, 250 and 500 μmol/L UA groups (96.5±20.1,115.5±30. 0, 152.5±26.7, 205.0±48. 2 and 230.1±22.5) compared with 6-OHDA group (F=10. 72, P< 0.05). Furthermore, there was a positive relationship of TH~+ neuron number with concentration of UA (F=13.94, P<0.05). The rate of cell viability of neurons in 5, 50, 100 and 500 μmol/L UA groups increased compared with 6-OHDA group (F= 40.96, P<0.05). There was no significant difference in cell viability of neurons between 250 μmol/L UA group and the control group (F=1.27, P>0.05). As compared with 6-OHDA group, the percentages of △ψm increased in UA groups (F= 8.82, P<0.05). The percentages of △ψm were higher in 100 and 250 μmol/L UA group than in control group (F = 4.62, P<0.05). Conclusions UA can reduce the neurotoxic effect of 6-OHDA, increase the viability of the cell and maintain mitochondrial membrane potential. The results suggest that UA shows a neuroprotective effect on dopaminergic neurons by anti-oxidative stress.
