Cystatin C敲除诱导的长爪沙鼠抑郁模型  

Knockout of Cystatin C Induced Depression Model in Gerbils

作　　者：伍颖 王诗媛 胡彩姣 朱筱 郭萌[1] 吕建祎[1] 刘欣[1] 李长龙[1] 霍学云[1] 陈振文[1] 杜小燕[1] WU Ying;WANG Shi-Yuan;HU Cai-Jiao;ZHU Xiao;GUO Meng;Lü Jian-Yi;LIU Xin;LI Chang-Long;HUO Xue-Yun;CHEN Zhen-Wen;DU Xiao-Yan(School of Basic Medical Sciences,Capital Medical University,Beijing 100069,China;Beijing Xuanwu Hospital,Capital Medical University,Beijing 100053,China)

机构地区：[1]首都医科大学基础医学院,北京100069 [2]首都医科大学北京宣武医院,北京100053

出　　处：《生物化学与生物物理进展》2022年第5期907-916,共10页Progress In Biochemistry and Biophysics

基　　金：国家自然科学基金(32070531);国家重点研发课题(2021YFF0702402)资助项目。

摘　　要：目的内源半胱氨酸蛋白酶抑制剂Cystatin C(CysC)由CST3基因编码,在脑中高表达,在多种神经病理过程中具有保护作用。本文旨在观察敲除(knock out,KO)长爪沙鼠CysC基因能否诱导动物的抑郁模型,并探讨模拟病理条件下CysC对内皮细胞和神经细胞的保护作用。方法使用qPCR验证CysC敲除(CysC-KO)沙鼠不同组织中CysC的转录水平;通过糖水偏好、社会互交、新物体识别、明暗箱和旷场实验,评估CysC-KO对长爪沙鼠行为的影响;分别在缺氧和饥饿(H&S)、氧糖剥夺/复氧(OGD/R)或炎症因子刺激(TNF-α/LPS)模拟的病理条件下,利用MTT法检测CysC或其抑制剂对人脐静脉内皮细胞(HUVEC)和小鼠神经母细胞瘤细胞(N2a)活力的影响。结果在CysC-KO长爪沙鼠的各种组织中CysC表达显著降低,尤其是大脑。长爪沙鼠的CysC缺失,可通过年龄依赖的方式诱导沙鼠的抑郁样行为,但其运动和探索行为未见异常。CysC在H&S、OGD/R和炎症条件下,能显著改善内皮细胞和神经细胞的增殖;而抑制CysC则具有相反作用。结论敲除CysC的长爪沙鼠表现出抑郁样行为,这可能是血管内皮细胞和神经元细胞失去CysC的保护作用引起。这些结果为研究CysC在抑郁发生中的作用机制提供了新的长爪沙鼠模型。Objective The endogenous cystatin C(CysC),encoded by the CST3 gene,is highly expressed in the brain and has protective effects in a variety of neuropathological processes.The aim of this research is to investigate whether knocking out(KO)CysC in gerbils can induce an animal model of depression and the protective effect of CysC on endothelial cells and neuronal cells under simulated pathological conditions.Methods The transcription level of CysC in different tissues of CysC knockout(CysC-KO)gerbils was detected using qPCR.The behavior of CysC-KO gerbils was evaluated through sucrose preference test(SPT),social interaction(SI),novel object recognition test(NOR),light/dark boxes,and open field trials(OFT).And under the pathological conditions simulated by H&S(hypoxic and starvation)or OGD/R(oxygen glucose deprivation/reoxygenation)or inflammatory factor(TNF-α/LPS),MTT assay was used to detect the effect of CysC or its inhibitor on HUVEC(human umbilical vein endothelial cells)and N2a(mouse neuroblastoma N2a cells)viability.Results CysC expression in CysC-KO gerbils was significantly reduced in various tissues,especially in the brain.Deficiency of CysC in gerbils induces depression-like behavior,but does not affect motion and exploration behavior.CysC significantly improved proliferation of both endothelial cell and neuronal cells under H&S,OGD and inflammatory conditions,while CysC inhibitor harbored contrary effect.Conclusion The CysC-KO gerbils exhibited depression-like behavior,which may be caused by the loss of the protective effect of CysC in vascular endothelial cells and neuronal cells.These results provide a new gerbil model for studying the neuroprotective effects of CysC and the mechanism of depression.

关 键 词：Cystatin C 长爪沙鼠 动物模型 抑郁行为 神经保护 

分 类 号：Q95[生物学—动物学]