叔丁基对苯二酚激活蛋白酶体活性促进成年小鼠神经干细胞增殖与分化  

Up-regulation of proteasome activity by tBHQ promotes the proliferation and differentiation of adult neural stem cells of mouse

作　　者：牛晓洁[1] 赵雪纯 崔佳彬 杨桂姣[1] 成晓龙[1] 陆利[1] 

机构地区：[1]山西医科大学人体解剖学教研室,太原030001 [2]山西医科大学临床一系,太原030001

出　　处：《神经解剖学杂志》2016年第5期641-646,共6页Chinese Journal of Neuroanatomy

基　　金：山西省自然科学基金(2015011132);山西省回国留学人员科研资助项目(2014-033);国家级大学生创新创业训练项目(201510114005)

摘　　要：目的:探讨叔丁基对苯二酚(tBHQ)对成年小鼠神经干细胞(aNSCs)增殖分化能力的影响,寻找维持aNSCs活力的有效手段。方法:成年BALB/c小鼠腹腔注射tBHQ(每日16.7 mg/kg)7 d,收集室管膜下区(SVZ)的脑组织,应用荧光酶标仪定量检测其蛋白酶体的活性;同时小鼠腹腔注射5-溴脱氧尿嘧啶核苷(BrdU),Brd U免疫荧光染色检测SVZ内a NSCs的增殖能力。分离培养SVZ aNSCs,tBHQ(20μmol/L)作用7 d,分析比较tBHQ组与DMSO组a NSCs的蛋白酶体活性、BrdU阳性率以及形成神经球的数量和直径。应用1%的胎牛血清诱导a NSCs分化,早期神经元标记物β-Ⅲ微管蛋白(Tuj1)免疫荧光染色并比较tBHQ组与DMSO组a NSCs向神经元分化的能力。结果:小鼠腹腔注射tBHQ,其SVZ蛋白酶体活性较DMSO对照组上调12.9%±4.6%(P<0.05),并且BrdU标记实验显示tBHQ组阳性细胞数为31.3±6.3,与DMSO对照组(15.4±1.3)比较,aNSCs增殖能力显著提高(P<0.05)。体外实验结果也显示tBHQ组aNSCs蛋白酶体活性较DMSO对照组上升10.1%±0.8%(P<0.01)。tBHQ组Brd U阳性率(31.3%±3.2%)是DMSO对照组(20%±1.5%)的1.6倍(P<0.05)。tBHQ组神经球的数量和直径分别是DMSO对照组的1.7倍(P<0.05)和1.4倍(P<0.05),提示tBHQ促进aNSCs的自我更新。此外,tBHQ组Tuj1阳性率为26.5%±1.6%,较DMSO对照组18.6%±2.1%显著提高(P<0.05),提示tBHQ能够促进a NSCs向神经元方向分化。结论:tBHQ能够上调蛋白酶体活性,促进aNSCs的增殖与分化。Objective： To investigate the effect of tBHQ on the proliferation and differentiation of adult neural stem cells（ aNSCs） of mouse and to find the effective way of maintaining aNSCs vitality. Methods： Adult BABL / c mice were injected intraperitoneally with tBHQ（ 16. 7 mg / kg / d） for 7 d. The brain tissue of subventricular zone（ SVZ） was collected to measure quantitatively the proteasomal activity by fluorescence microplate. 5-Bromo-2＇-deoxy Uridine（ BrdU） labeling was used to detect the proliferating cells in SVZ by immunofluorescence staining. After incubation with 20μmol / L tBHQ for 7 d in vitro,the proteasome activity of aNSCs in culture,the percentage of BrdU＋cell as well as the diameter and quantity of neurospheres were analyzed in tBHQ group with respect to DMSO control group. Meanwhile,the immunostaining of Tuj1,a new neuron marker,was examined to indicate neural differentiation of aNSCs induced by1% serum in tB HQ- and DMSO- group. Results： After the injection of tB HQ in abdominal cavity,the proteasomal activity of SVZ was increased 12. 9% ± 4. 6%（ P〈0. 05）. The number of BrdU ＋cells was significantly increased from15. 4 ± 1. 3（ DMSO group） to 31. 3 ± 6. 3（ tB HQ group） by BrdU labeling. It showed that the proliferation of aN SCs increases dramaticlly（ P〈0. 05）. After aN SCs were cultured in vitro with tB HQ for 7d,the proteasomal activity of aN SCs increased by 10. 1 ± 0. 8%（ P〈0. 01）. The BrdU ＋rate of tB HQ group（ 31. 3% ± 3. 2%） was 1. 6 times of DMSO group（ 20% ± 1. 5%）（ P〈0. 05）. The number and diameter of neurospheres in tB HQ group were 1. 7（ P〈0. 05） and 1. 4（ P〈0. 05） times as much as DMSO group respectively. It indicated that tB HQ could accelerate the ability of self-renewal of aN SCs. After differentiation the percentage of Tuj1＋was obviously increased from 18. 6% ±2. 1%（ DMSO group） to 26. 5% ± 1. 6%（ tBHQ group,P〈0. 05）. It revealed that tB HQ could promote aN SCs to differentiate into

关 键 词：叔丁基对苯二酚 成年神经干细胞 增殖 分化 小鼠 

分 类 号：R741[医药卫生—神经病学与精神病学]