老化相关的自发荧光可作为细胞衰老的标志物

发布时间：2018-11-07 18:36

【摘要】：目的观察衰老的脑组织内自发荧光的波长特点,探讨如何合理地利用和消减老化相关的自发荧光。方法选取3月龄和24月龄自然老化的C57BL/6J雌、雄性小鼠。以衰老相关β-半乳糖苷酶染色(SA-β-Gal)和脂褐素的沉积评估老年鼠脑组织衰老的程度。以激光共焦显微镜分析老化相关自发荧光的波长特点,免疫组织化学和免疫荧光的方法标记星形胶质细胞。结果老年鼠海马和下丘脑第三脑室室周旁出现强SA-β-Gal染色、脂褐素的聚集及自发荧光。老化相关的自发荧光发射波长介于500~565nm,下丘脑第三脑室室周旁的星形胶质细胞不仅高表达SA-β-Gal,而且出现很强的自发荧光。采用针对脂褐素的自发荧光消除剂处理脑组织后,老化相关的自发荧光几乎完全消失。结论老化相关的自发荧光可以作为特定表型细胞衰老的标志物;恰当的处理老化相关的自发荧光,可极大的提高衰老脑组织免疫荧光检测的特异性。
[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.
【作者单位】：福建省高校脑老化与神经变性疾病重点实验室福建省老年医学研究所;福建省分子神经病学重点实验室福建医科大学附属协和医院神经内科;
【基金】：国家重点基础研究发展计划973课题资助项目(2007CB507404) 福建省教育厅科技资助项目(JK2010025)
【分类号】：R329

【共引文献】