衰老对帕金森病模型小鼠线粒体复合物活性的影响

目的探讨衰老对帕金森病(PD)模型小鼠线粒体复合物的影响。方法选用8个月龄、16个月龄雌性C57BL/6小鼠,随机分为8个月龄对照组、8个月龄模型组、16个月龄对照组及16个月龄模型组。模型组小鼠皮下注射1-甲基-4-苯基-1,2,3,6-四氢吡啶(MPTP)制成PD模型,对照组注射等量生理盐水;进行行为学测试,并检测其黑质多巴胺含量,采用紫外分光光度法检测线粒体复合物Ⅰ~Ⅳ的活性。结果与对照组相比,模型组小鼠行为学异常,黑质多巴胺含量显著下降(P<0.05),线粒体复合物Ⅰ的活性明显低于对照组(P<0.05);与8个月龄PD小鼠相比,16个月龄小鼠行为学成绩、多巴胺含量及线粒体复合物Ⅰ的活性降低更明显(P<0.05)。各组线粒体复合物Ⅱ、Ⅲ、Ⅳ无显著差异。结论小鼠黑质纹状体系统线粒体复合物Ⅰ功能障碍可能是导致PD发生的重要原因之一,衰老可加速其功能障碍。     

雷帕霉素对帕金森病小鼠模型线粒体呼吸功能及跨膜电位的影响

目的探讨雷帕霉素(RAPA)对帕金森病(PD)小鼠模型线粒体呼吸功能及其跨膜电位的影响。方法将54只昆明小鼠随机分为3组:空白组,模型组,用药组。空白对照组不进行任何处理;模型组与用药组均采用皮下注射1-甲基-4-苯基四氢吡啶(MPTP)法制作PD模型;用药组在制备模型前、中、后持续给予RAPA灌胃给药,空白组与模型组同时给予同体积的生理盐水。给药结束后对各组小鼠采用自主活动及滚轴运动进行行为学评分,高效液相-电化学法(HPLC-ECD)检测黑质DA含量,比色法定量检测线粒体复合物Ⅰ活性,应用流式细胞仪分析线粒体的跨膜电位。结果与空白组比较,模型组、用药组的行为学成绩、黑质多巴胺水平、复合物Ⅰ活性、跨膜电位均显著下降(P<0.05);而与模型组比较,给用药组的各项指标均显著改善(P<0.05)。结论 RAPA可以有效改善PD小鼠模型行为学表现,其机制可能与改善线粒体膜电位和线粒体复合物活性有关。     

线粒体跨膜电位在不同年龄帕金森病大鼠模型的改变

目的探讨线粒体跨膜电位在不同年龄帕金森病(PD)模型大鼠黑质纹状体系统神经细胞中的改变。方法选用15周龄、53周龄雄性Wistar大鼠,随机分为15周龄对照组、15周龄模型组、53周龄对照组和53周龄模型组。采用立体定位技术双点注射6-羟基多巴胺(6-OHDA)制成PD模型,对照组注射等体积无菌生理盐水。皮下注射阿扑吗啡(APO)进行旋转实验,流式细胞仪检测各组大鼠黑质细胞线粒体膜电位的改变。结果与对照组相比,模型组大鼠行为学异常,纹状体多巴胺含量显著下降(P<0.05),线粒体膜电位明显低于对照组(P<0.05);与15周龄PD大鼠相比,53周龄PD大鼠的行为学成绩、多巴胺含量及线粒体膜电位的活性降低更明显(P<0.05)。结论 6-OHDA所致的PD模型大鼠黑质纹状体系统线粒体膜电位下降,可能是导致PD发生发展的机制之一;老化可加速其功能障碍。     

沉默信息调节因子1/p53信号通路参与MPTP诱导的帕金森病小鼠多巴胺能神经元丢失

目的研究沉默信息调节因子1(SIRT1)和p53在MPTP诱导的帕金森病(PD)小鼠模型多巴胺能神经元凋亡中的可能作用。方法将健康雄性C57BL/6小鼠随机分为对照组、MPTP组,采用行为学方法检测行为学改变,高效液相色谱(HPLC)检测多巴胺(DA)、二羟基苯乙酸(DOPAC)和高香草酸(HVA)的含量变化,免疫荧光染色法观察两组小鼠黑质酪氨酸羟化酶(TH)阳性神经元数目的变化及SIRT1表达情况,TUNEL法观察黑质细胞凋亡情况,Western blot法检测TH、SIRT1、p53、乙酰化p53 (ac-p53)、B淋巴细胞瘤-2基因(Bcl-2)和Bax的表达情况。结果行为学结果显示MPTP组小鼠爬杆转向时间及爬杆总时间均较对照组小鼠显著延长(P 0. 01)。HPLC结果提示MPTP组的DA、DOPAC及HVA含量较对照组显著下降(P 0. 01)。免疫荧光结果显示MPTP小鼠黑质区TH阳性神经元数目及SIRT1表达较对照组均显著减少。TUNEL检测结果显示,与对照组相比,MPTP组凋亡阳性细胞数明显增多。Western blot结果显示,与对照组相比,MPTP组的TH、SIRT1、Bcl-2蛋白表达显著下降(P 0. 01),p53、ac-p53、Bax蛋白表达显著升高(P 0. 01)。结论MPTP模型小鼠行为学异常、TH阳性神经元减少、DA及其代谢产物下降提示成功复制PD动物模型,同时MPTP模型小鼠的SIRT1、p53及凋亡相关蛋白表达异常,提示该信号通路可能参与了PD的疾病过程。     

氯化锂对MPTP致帕金森病小鼠模型行为学及黑质多巴胺能神经元影响的研究

目的 探讨氯化锂对MPTP致帕金森病小鼠模型行为学及黑质多巴胺能神经元保护作用的影响.方法 实验小鼠随机分为MPTP组、NS(生理盐水)组、LM(LiCl +MPTP)组、PM(PBS+ MPTP)组;通过行为学观察各组震颤麻痹、移动格子数、站立次数、滚轴次数、游泳能力;免疫组织化学染色与免疫印迹技术观察各组酪氨酸羟化酶(TH)和钙结合蛋白(CB)的表达变化.结果 行为学检测LM组震颤麻痹评分、移动格子数、站立次数、滚轴实验、游泳能力都显著高于PM组(P＜0.05);免疫组织化学染色结果显示LM组黑质致密部TH与CB阳性神经元数量显著多于PM组;Western blot免疫印迹结果显示LM组TH、CB蛋白含量表达水平显著高于PM组(P＜0.01或0.05).结论 氯化锂可明显改善MPTP所致小鼠PD模型的运动障碍,且对其引起的DA神经元损伤起到保护作用,这种保护作用与细胞内CB表达增加有关;PD模型行为学与多巴胺能神经元的变化具有一致性.     

尼莫地平对帕金森病模型鼠黑质多巴胺能神经元中Bcl-2、P53蛋白表达的影响

目的观察尼莫地平对帕金森病模型鼠多巴胺能神经元中Bcl-2、P53蛋白表达的影响,从而探索尼莫地平对黑质多巴胺能神经元的保护作用。方法建立大鼠PD模型,分组、分阶段用尼莫地平进行干预,第一阶段为尼莫地平对PD模型的预先干预,第二阶段为成功PD模型的药物治疗(尼莫地平、左旋多巴),其后均由阿朴吗啡(Apomorphine)诱导旋转行为,最后予大鼠黑质细胞进行HE、TH、Bcl-2、P53染色。结果第一阶段:尼莫地平PD模型组(Ⅰ组)和PD模型组(Ⅱ组),成功模型右侧黑质Bcl-2蛋白表达阳性细胞百分比较假手术组(Ⅲ组)和正常对照组(Ⅳ组)低(P<0.05),而P53蛋白表达阳性细胞百分比较Ⅲ组和Ⅳ组高(P<0.05);Ⅰ组右侧黑质Bcl-2蛋白表达阳性细胞百分比高于Ⅱ组(P<0.05),而P53蛋白表达阳性细胞百分比低于Ⅱ组(P<0.05);第二阶段:尼莫地平组、左旋多巴组或二者联用干预组与生理盐水组间右侧黑质Bcl-2、P53蛋白表达阳性细胞百分比无统计学差异(P>0.05)。结论尼莫地平在蛋白合成水平促进了Bcl-2表达、抑制了P53表达,减缓了多巴胺能神经元的凋亡。     

还原型谷胱甘肽对帕金森病保护性治疗的实验研究

目的观察还原型谷胱甘肽对帕金森病(PD)大鼠模型的保护性治疗作用,并探讨其作用机理.方法应用6-羟基多巴制作PD大鼠模型.将大鼠模型随机分为4组(每组8只):模型组,谷胱甘肽(GSH)组,左旋多巴(Ldopa)组,谷胱甘肽 左旋多巴组,另设对照组(8只),分别给予相应处理,共45天,给药前后均进行行为学测试,给药结束后行免疫组化和电镜观察,并测定黑质区谷胱甘肽过氧化物酶(GSH-Px)、丙二醛(MDA)、活性氧(ROS)及线粒体呼吸链酶复合体Ⅰ水平.结果 (1)GSH对其旋转行为无明显影响(P＞0.05);(2)GSH 能减轻黑质区氧化应激损伤;(3)GSH能增强黑质呼吸链酶复合体Ⅰ活性;(4)免疫组化发现GSH组TH-IR神经元较模型组明显增多(P＜0.001);(5)电镜下发现GSH可部分延缓凋亡进程.结论 (1)GSH能减轻黑质区氧化应激损伤,并对线粒体呼吸链具有一定保护作用;(2)GSH与L-dopa合用既可有效改善症状,又对残存黑质多巴胺能神经元具有保护性治疗作用.     

帕金森病模型大鼠黑质多巴胺能神经元的氧化应激研究

目的　探索帕金森病模型大鼠黑质多巴胺能神经元的氧化应激发病机制。方法　通过立体定位仪 ,将 6-OHDA注入大鼠一侧纹状体内制备 PD模型 ,2周后观察动物的行为学改变 ,2个月后观察黑质纹状体等的病理形态学变化 ,检测模型组、假手术组和正常对照组的超氧化物歧化酶 (SOD)的活性 ,丙二醛 (MDA)、一氧化氮(NO)代谢产物 (NO x)的含量的变化。结果　成功 PD模型鼠有 2 2只。光镜下 HE染色示模型组右侧黑质的多巴胺神经元受损 ,数目减少。模型组右侧黑质的 SOD的含量下降 ,MDA及 NO x含量明显升高 ,与左侧、假手术组及正常对照组相比有显著差异 (P>0 .0 5)。结论　 6-OHDA纹状体内双靶点注射法是一种有效的制备 PD模型的方法。帕金森病大鼠模型黑质内 SOD活性下降 ,MDA、NO x含量升高 ,氧化应激在 PD的发病中起重要的作用     

慢性间断性缺氧对帕金森病模型小鼠行为学及黑质病理改变的影响

目的：探讨慢性间断性缺氧（CIH）对百草枯诱导的帕金森病（PD）模型小鼠行为学及黑质病理改变的影响。方法：雄性6周龄C57BL／6小鼠44只,随机分为对照组、百草枯组、CIH组和百草枯＋CIH组,通过爬杆实验和自主活动记数检测小鼠行为学变化;苏木精-伊红染色及焦油紫染色和电镜观测黑质多巴胺能神经元形态学及细胞数的变化。结果：①百草枯＋CIH组较其余3组爬杆实验评分、自主活动计数明显减少;②百草枯组、CIH组和百草枯＋CIH组黑质多巴胺能神经元与对照组比较,均出现不同程度的形态学改变,以百草枯＋CIH组最为严重;焦油紫染色：对照组、CIH组、百草枯组和百草枯＋CIH组黑质细胞计数分别为70．09±6．46、38．36±4．34、53．09±4．55和23．18±4．85,3组黑质多巴胺能神经元与对照组比较均出现不同程度的减少,且以百草枯＋CIH组减少最为明显（P〈0．01）。结论：CIH加重了百草枯所致PD模型小鼠行为学及黑质多巴胺能神经元的病理损害。     

左旋多巴对帕金森病大鼠毒性作用的实验研究

目的　研究左旋多巴 (L dopa)对帕金森病 (PD)模型大鼠异常行为、黑质抗氧化系统、线粒体呼吸链功能和神经递质代谢的影响及其机制。方法　应用 6 羟基多巴胺 (6 OHDA)立体定向注射制作PD大鼠模型 ,给PD大鼠L dopa 2 5mg/ (kg·d)灌胃 ,共 4 5d。给药前后分别进行行为学测试 ,给药后测定黑质区谷胱甘肽过氧化物酶 (GSH Px)、丙二醛 (MDA)、活性氧 (ROS)及线粒体呼吸链酶复合体Ⅰ水平 ,测定尾状核头部多巴胺 (DA)、高香草酸 (HVA)、单胺氧化酶 B(MAO B)的水平。结果　 (1)L dopa组大鼠旋转速度给药前为(13.1± 1.5 )r/min ,给药后为 (7.2± 1.6 )r/min,给药前后比较差异有显著性 (P <0 .0 1) ;(2 )L dopa组GSH Px活性、呼吸链酶复合体Ⅰ水平降低 ,MDA含量、ROS活性升高 ,与对照组比较差异均有显著性 (均P <0 .0 1) ;(3)L dopa组MAO B活性、DA、HVA含量及DA/HVA比值与对照组比较均显著升高 (P <0 .0 5～ 0 .0 1)。结论L dopa能有效改善PD大鼠的旋转行为 ,但可加重黑质区氧化应激损伤 ,抑制线粒体呼吸链酶活性。     

“抗帕颗粒”对帕金森病模型鼠的神经保护作用

目的探讨"抗帕颗粒"对1-甲基-4-苯基-1,2,3,6-四氢吡啶(MPTP)帕金森病(PD)模型小鼠黑质纹状体区TH阳性神经元及多巴胺(DA)的影响。方法 90只健康雄性C57BL/6小鼠,鼠龄8~12w,随机分为3组:正常对照组30只、PD模型对照组30只、PD模型干预组30只;MPTP腹腔注射(40mg·kg~(-1)·d~(-1)×7)制备小鼠PD模型;正常对照组及PD模型对照组予生理盐水1m L·d~(-1)灌胃,PD模型干预组给予"抗帕颗粒"40mg·kg~(-1)·d~(-1)灌胃,连续喂养4个月。比较分析各组4月时黑质纹状体区TH阳性神经元及DA情况。结果 1正常对照组30只(30/30只)最终均存活,PD模型对照组4个月时存活27只(27/30只),PD模型干预组4个月时存活28只(28/30只);2PD模型对照组、PD模型干预组小鼠每次注射MPTP后,先有短暂兴奋(持续7.61±2.17min),表现为四处窜跳;随即出现全身中重度震颤,皮毛及尾巴时有竖立,活动减少,持续24.23±3.89min后震颤消失;随后出现活动减少;3经Imagepro~Plus 5.1系统分析,正常对照组TH阳性细胞面积为64145μm~2,高倍镜下可见大量胞质为褐色颗粒的TH阳性细胞;PD模型对照组TH阳性细胞染色面积为40012μm~2,高倍镜下见TH细胞数量明显减少;PD模型干预组TH阳性细胞染色面积为60952μm~2,高倍镜下见TH阳性细胞数量较PD模型对照组增加;4正常对照组DA含量为2.18±0.31μg·m L~(-1),与PD模型对照组1.57±0.22μg·m L~(-1)比较,P0.01;正常对照组与PD模型干预组2.04±0.18μg·m L~(-1)比较。P0.05;PD模型对照组与PD模型干预组比较,P0.01。结论 "抗帕颗粒"可使PD小鼠黑质纹状体中多巴胺能神经元一定程度地减少丢失,并改善DA含量的下降,对多巴胺能神经元的数量、形态及功能具有一定的保护作用,有望改善PD治疗现状并在临床进一步推广应用。     

α7神经型尼古丁受体激动剂PNU282987对APP/PS1小鼠海马组织中DYN-Ⅰ蛋白表达的影响

目的研究特异性激动APP/PS1转基因小鼠脑组织中α7神经型尼古丁受体(α7 nAChR)水平后对海马组织DYN-Ⅰ蛋白的影响;探讨α7 nAChR在阿尔茨海默病(Alzheimer disease,AD)发病机制中的神经保护作用机制。方法实验动物分为对照组(Control)、野生加PNU282987组(WP)、APP/PS1转基因组(APP/PS1)和APP/PS1加PNU282987组(AP)各8只,WP和AP组给予α7 nAChRs特异性激动剂PNU-282987,另两组给予生理盐水,给药方式为小鼠24 w龄时1 mg/kg腹腔注射PNU-282987,连续5 d。采用Real-time PCR法和蛋白免疫印迹(Western Blot)法分别测定小鼠海马组织中发动蛋白Ⅰ(DYN-Ⅰ)mRNA和蛋白表达水平的变化。结果与control组相比,APP/PS1组海马组织中发动蛋白Ⅰ(DYN-Ⅰ)mRNA和蛋白水平下降(P0.05,P0.01);而特异性激动α7 nAChR水平后,与对照组相比WP组中发动蛋白Ⅰ(DYN-Ⅰ)mRNA和蛋白水平升高(P0.05,P0.01);与APP/PS1组相比AP组小鼠大脑海马组织中发动蛋白Ⅰ(DYN-Ⅰ)mRNA和蛋白水平明显升高(P0.01,P0.01)。结论特异性激动APP/PS1转基因小鼠海马组织中α7 nAChR水平能够使网格蛋白内吞调节蛋白DYN-Ⅰ表达升高。这可能提示了α7 nAChR对突触有一定的保护作用,进一步说明α7 nAChR在阿尔茨海默病的发病中起着重要作用。     

Increased vulnerability with aging to MPTP: the mechanisms underlying mitochondrial dynamics

Abstract

Objective:

The risk and vulnerability of Parkinson disease (PD) are especially high in the elderly. However, the underlying causes are unknown. 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP) mice are useful tools to some extent for investigating PD-related problems due to their PD-like symptoms. The study is aimed to determine whether and what mitochondrial-events during aging are related with the increased vulnerability of the elderly to MPTP.

Methods:

The MPTP mice were established by intraperitoneal injection of MPTP, control animals were injected with saline. Mice of different ages were divided into six groups: 4-month old control group, 4-month old MPTP group, 8-month old control group, 8-month old MPTP group, 15-month old control group, and 15-month old MPTP group. The behavior ability and pathology were assessed for each mouse. Mitochondrial functions were measured and compared among different groups. Mitochondrial fusion/fission-related proteins and autophagic proteins were analyzed by western blotting.

Results:

The elder animals were impaired more severely in behavior and pathology than the young ones. Then, we revealed that aging is associated with the degeneration of several mitochondrial functions, disturbed balance of fusion/fission as well as decreased activity of autophagic proteins. More importantly, mitochondria in the elderly are more vulnerable to MPTP treatment than that in the young, which may contribute to the increased risk and vulnerability of the elderly to MPTP.

Conclusion:

We identified several changes of mitochondrial-events with aging MPTP mice that could be related to the MPTP susceptivity and vulnerability, which would provide significant instructions for future in developing proper interventions that lower the vulnerability, or slow the progression of PD in the elderly.     

“抗帕颗粒”对帕金森病模型鼠α-syn异常聚集的影响

目的探讨"抗帕颗粒"对1-甲基-4-苯基-1,2,3,6-四氢吡啶(MPTP)帕金森病(PD)模型小鼠黑质纹状体区α-突触共核蛋白(α-syn)异常聚集的影响。方法 90只健康雄性C57BL/6小鼠,鼠龄8~12w。随机分为3组,正常对照组30只、PD模型对照组30只、PD模型干预组30只。MPTP腹腔注射40mg·Kg-1·d-1×7制备小鼠PD模型;正常对照组及PD模型对照组予生理盐水1ml·d-1灌胃;PD模型干预组给予"抗帕颗粒"40mg·kg-1·d-1灌胃,连续喂养4个月。比较分析各组4月时黑质纹状体区α-syn聚集情况。结果 (1)正常对照组30只(30/30只)最终均存活,PD模型对照组4个月时存活27只(27/30只),PD模型干预组4个月时存活28只(28/30只);(2)PD模型对照组、PD模型干预组小鼠每次注射MPTP后,先有短暂兴奋(持续7.61±2.17min),表现为四处窜跳;随即出现全身中重度震颤,皮毛及尾巴时有竖立,活动减少,持续24.23±3.89min后震颤消失;随后出现活动减少;(3)黑质纹状体区α-syn免疫荧光双染检测;(3)PD模型对照组较正常对照组明显增多,PD模型干预组较PD模型对照组有所减少,但仍较正常对照组增多;(4)Western blotting检测α-syn蛋白量的表达,正常对照组与PD模型对照组比较,P0.001;与PD模型干预组比较,P0.001;PD模型对照组与PD模型干预组比较,P0.05。结论 "抗帕颗粒"对PD小鼠黑质纹状体区α-syn异常聚集具有一定程度的抑制作用,阻断氧化应激反应可能是这种保护作用的关键机制,有望改善PD治疗现状并在临床进一步推广应用。     

Chronic levodopa administration alters cerebral mitochondrial respiratory chain activity

Parkinson's disease (PD) is characterized mainly by a loss of nigrostriatal dopamine neurons. Thus far, the actual physiopathology of PD remains uncertain, although recent studies have found decreased activity of complex I, one of the enzymatic units of the mitochondrial respiratory chain, in various tissues of PD patients. Because most, if not all, of PD patients are treated chronically with levodopa, the precursor of dopamine, and because we have shown previously that catecholamines may alter mitochondrial respiration, we assessed the effects of chronic administration of levodopa on complex I activity in rat brain. We found that chronic administration of levodopa, at a dose used in PD patients, caused a significant reduction in complex I activity while it did not affect the activities of complex II, complex IV, and citrate synthase. Reduction in complex I activity correlated well with catecholamine innervation as the reduction was observed mainly in the striatum and substantia nigra and to a lesser extent in the frontal cortex but not in the cerebellum. Moreover, the levodopa-induced decrease of complex I activity was reversible since activities at 1, 3, and 7 days after the last injection showed a progressive return to control values. Incubation of whole brain mitochondria in vitro showed that both levodopa and dopamine inhibit complex I activity in a dose- and time- dependent manner. In contrast, other compounds such as homovanillic acid, 3,4-dihydroxyphenylacetic acid, and 3-O-methyl-dopa were minimally effective. Reduced glutathione, ascorbate, superoxide dismutase, and catalase prevented the effect of levodopa and dopamine on complex I. Various inhibitors of monoamine oxidase also prevented the effect of dopamine. In agreement with a critical role of monoamine oxidase in this effect in vitro, we observed that noncatecholamine substrates of this enzyme such as serotonin and β-phenylethylamine were potent inhibitors of complex I. However, autoxidation may also be involved in this process because the effect of levodopa, 6-hydroxydopa, and 6-hydroxydopamine on complex I activity was only partially suppressed by monoamine oxidase inhibitors. These observations demonstrate that the chronic administration of levodopa can cause alterations in mitochondrial respiratory chain activity in rats that are most likely related to an oxidative stress provoked by the increase in dopamine turnover. We suggest that this mechanism may exaggerate a mitochondrial defect already present in the brains of PD patients and thus may play a role in the progression of PD.     

The effects of rehabilitative training on the behaviors of Parkinson's disease mouse and underlying mechanism

ABSTRACT Objective: To examine the effects of rehabilitative training on mice behavior improvement in a 1-Methyl-4-Phenyl-1,2,3,6-Tetrahydropyridine (MPTP) mice model of Parkinson's disease (PD). Methods: MPTP mice model of Parkinson disease was generated. The rehabilitative training included daily running wheel and balance walking; the control group was allowed for free activity. The behaviors of the two groups were investigated at 1 week, 2 weeks, and 4 weeks and the content of dopamine (DA) in striatum of different groups of mice was measured with high-performance liquid chromatography (HPLC). Results: The results showed that as compared to the control group, the score of climbing pole, traction, swimming tests, and dopamine content was improved in the rehabilitative training group. Conclusion: This suggested that rehabilitative training could improve the locomotor ability and dopamine content in PD mouse model.     

Mitochondrial energy metabolism changes during aging-mouse cranial nerve cells treated with various doses and forms of Fructus schizandrae

BACKGROUND： During the cellular aging process, the number of mitochondria, generation of adenosine triphosphate （ATP）, activity of respiratory chain enzyme complex 1 and 4, and oxidation decrease. OBJECTIVE： To observe the effects of aqueous and spirituous extract, as well as polysaccharides from Fructus schizandrae （Magnolia Vine） on energy metabolism and mitochondrial anti-oxidation in cranial nerve cells of a D-gal-induced aging mouse model. DESIGN, TIME AND SETTING： A randomized, controlled, animal study. The experiment was conducted at the Department of Biochemistry, Qiqihar Medical College between March and July 2006. MATERIALS： Fifty healthy, Kunming mice of both sexes, aged 2 3 months old and weighing 18 22 g, were used for the present study. Fructus schizandrae was purchased from the Medical College of Jiamusi University. Aqueous extracts, spirituous extracts, and polysaccharides from Fructus schizandrae were prepared. D-galactose （D-gal） is a product of the Second Reagent Factory, Shanghai City, China. Mn-superoxide dismutase （Mn-SOD） kit, malonaldehyde （MDA） kit, protein quantification kit, and inorganic phosphorus testing kit were purchased from Jian Cheng Bioeng. Co., China. METHODS： Fifty mice were randomly divided into five groups, with 10 mice in each group： young control, aging model, aqueous Fructus schizandrae extract, spirituous Fructus schizandrae extract, and Fructus schizandrae polysaccharides. Over a course of 30 days, mice in aging model, aqueous Fructus schizandrae extract, spirituous Fructus schizandrae extract, and Fructus schizandrae polysaccharides groups were injected subcutaneously with D-gal （100 mg/kg） into the nape of the neck daily, and administered intragastrically with an equal volume of sterile, warm water （aging model）, aqueous Fructus schizandrae extract （2 g/kg）, spirituous Fructus schizandrae extract （2 g/kg）, or Fructus schizandrae polysaccharides （0.2 g/kg）, respectively. Mice in the young control group were injected     

LRRK2干扰对MPP+诱导PD细胞模型线粒体功能的影响

目的 探讨富亮氨酸重复激酶2(LRRK2)干扰对1-甲基-4-苯基吡啶离子(MPP+)诱导的帕金森病(PD)细胞模型线粒体功能及CaMKK-β/AMPK通路的影响.方法 采用MPP+诱导传代培养的SH-SY5Y细胞构建PD细胞模型.将造模的SH-SY5Y细胞随机分PD模型组、空载转染组(空载转染PD模型细胞)和siRN...     

晚期糖基化终末产物受体对帕金森病模型小鼠脑内酪氨酸羟化酶表达的影响

目的探讨晚期糖基化终末产物受体(RAGE)对1-甲基-4-苯基-1,2,3,6-四氢吡啶(MPTP)诱导的帕金森病(PD)模型小鼠脑中酪氨酸羟化酶(TH)表达量的影响。方法采用12周龄的C57BL/6雄性小鼠依据不同处理方法分为6组(均n=10):对照组;MPTP组;空载病毒阴性对照(RAGE-NC)组;目的基因阴性对照(siRNA-RAGE)组;RAGE-NC+MPTP组;siRNA-RAGE+MPTP组。携带空载siRNA的慢病毒(RAGE-NC)与携带抑制RAGE表达的目的 siRNA-RAGE慢病毒经脑立体定位仪定向注射于小鼠两侧黑质,根据分组情况给予腹腔注射MPTP 30mg·kg~(-1)或等量生理盐水(每周2次×5周)。5周后予小鼠断头取脑,利用免疫组织荧光染色法检测各组小鼠脑组织黑质中TH数量变化情况,采用Western blot法分别检测各组RAGE、caspase-3、TH蛋白表达水平。结果与RAGENC+MPTP组比较,siRNA-RAGE+MPTP组RAGE蛋白表达减少(0.782 8±0.139 6 vs 1.039 0±0.146 4,P0.01),caspase-3蛋白表达减少(0.864 4±0.105 3 vs 1.240 0±0.080 7,P0.001),TH蛋白表达量显著升高(1.114 0±0.201 1vs 0.771 1±0.211 3,P0.05),差异有统计学意义。结论抑制RAGE表达可抑制凋亡反应,提高PD多巴胺能神经元模型中TH蛋白表达水平,有潜在的神经保护作用。