【摘要】：目的觀察衰老的腦組織內(nèi)自發(fā)熒光的波長特點(diǎn),探討如何合理地利用和消減老化相關(guān)的自發(fā)熒光。方法選取3月齡和24月齡自然老化的C57BL/6J雌、雄性小鼠。以衰老相關(guān)β-半乳糖苷酶染色(SA-β-Gal)和脂褐素的沉積評(píng)估老年鼠腦組織衰老的程度。以激光共焦顯微鏡分析老化相關(guān)自發(fā)熒光的波長特點(diǎn),免疫組織化學(xué)和免疫熒光的方法標(biāo)記星形膠質(zhì)細(xì)胞。結(jié)果老年鼠海馬和下丘腦第三腦室室周旁出現(xiàn)強(qiáng)SA-β-Gal染色、脂褐素的聚集及自發(fā)熒光。老化相關(guān)的自發(fā)熒光發(fā)射波長介于500~565nm,下丘腦第三腦室室周旁的星形膠質(zhì)細(xì)胞不僅高表達(dá)SA-β-Gal,而且出現(xiàn)很強(qiáng)的自發(fā)熒光。采用針對(duì)脂褐素的自發(fā)熒光消除劑處理腦組織后,老化相關(guān)的自發(fā)熒光幾乎完全消失。結(jié)論老化相關(guān)的自發(fā)熒光可以作為特定表型細(xì)胞衰老的標(biāo)志物;恰當(dāng)?shù)奶幚砝匣嚓P(guān)的自發(fā)熒光,可極大的提高衰老腦組織免疫熒光檢測的特異性。
[Abstract]:Objective to observe the wavelength characteristics of autofluorescence in aging brain tissue and to explore how to utilize and subtract the autofluorescence related to aging. Methods female and male C57BL/6J mice aged 3 months and 24 months were selected. Aging related 尾 -galactosidase staining (SA- 尾-Gal) and lipofuscin deposition were used to evaluate the aging degree of senile brain tissue. The wavelength characteristics of autofluorescence associated with aging were analyzed by laser confocal microscopy, and astrocytes were labeled by immunohistochemistry and immunofluorescence. Results strong SA- 尾-Gal staining, lipofuscin aggregation and autofluorescence were observed in the hippocampus and the hypothalamic periventricular area of the aged rats. Aging related autofluorescence emission wavelength was between 500 and 565 nm. Astrocytes in hypothalamic periventricular region not only overexpressed SA- 尾-Gal, but also showed strong autofluorescence. The aging related autofluorescence almost disappeared after the brain tissue was treated with lipofuscin-specific autofluorescence eliminator. Conclusion Aging related autofluorescence can be used as a marker of specific phenotypic cell senescence, and the specificity of immunofluorescence detection in aging brain tissue can be greatly improved by properly dealing with aging related autofluorescence.
【作者單位】： 福建省高校腦老化與神經(jīng)變性疾病重點(diǎn)實(shí)驗(yàn)室 福建省老年醫(yī)學(xué)研究所;福建省分子神經(jīng)病學(xué)重點(diǎn)實(shí)驗(yàn)室 福建醫(yī)科大學(xué)附屬協(xié)和醫(yī)院神經(jīng)內(nèi)科;
【基金】：國家重點(diǎn)基礎(chǔ)研究發(fā)展計(jì)劃973課題資助項(xiàng)目(2007CB507404) 福建省教育廳科技資助項(xiàng)目(JK2010025)
【分類號(hào)】：R329

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本文編號(hào)：2317238