组织蛋白酶K对大鼠空间学习记忆功能的影响*

doi:10.12047/j.cjap.6311.2022.077

中国应用生理学杂志 ›› 2022, Vol. 38 ›› Issue (5): 412-417.doi: 10.12047/j.cjap.6311.2022.077

组织蛋白酶K对大鼠空间学习记忆功能的影响^*

陈丽^1,2+, 陈金信³⁺, 王心怡^1,2, 李香兰⁴, 姜海英^2△

1.浙江中医药大学, 杭州 310000;
2.嘉兴学院, 嘉兴 314000;
3.三亚学院, 三亚 572000;
4.延边大学, 延吉 136200

收稿日期:2022-04-07 修回日期:2022-09-14 出版日期:2022-09-28 发布日期:2023-04-23
通讯作者: ^△Tel: 18457392565; E-mail: hyjiang7689@zjxu.edu.cn.
作者简介:⁺: 为共同第一作者
基金资助:
^*浙江省基础公益研究计划项目(LGJ20H090001);国家自然科学基金项目(81760207);三亚学院青年教师专项培养科研项目(USYQNZX21-21)

Effects of Cathepsin K on spatial learning and memory in rats

CHEN Li^1,2+, CHEN Jin-xin³⁺, WANG Xin-yi^1,2, LI Xiang-lan⁴, JIANG Hai-ying^2△

1. Zhejiang Chinese Medical University, Hangzhou 310000;
2. Jiaxing University, Jiaxing 314000;
3. University of Sanya, Sanya 572000;
4. Yanbian University, Yanji 136200, China

Received:2022-04-07 Revised:2022-09-14 Online:2022-09-28 Published:2023-04-23

摘要/Abstract

摘要： 目的: 探讨组织蛋白酶K(CatK)对大鼠海马空间学习和记忆功能的影响及其神经化学机制。方法: SD雄性大鼠随机分为对照组(Control组)和CatK阻断组(CatK Ⅱ组)每组各10只,分别于海马DG区微量注射组织蛋白酶K特异性阻断剂(0.5 μg/μl)和等量人工脑脊液5 d;原代培养海马神经元细胞,设对照组(CON组)、阴性对照组(NC组)和siRNA干扰组(siCatK组),每组设3个复孔,siRNA导入18～20 h后收集样本。应用Morris水迷宫评估大鼠空间学习和记忆功能,同时在大鼠清醒、自由活动状态下,结合脑部微量透析法和高效液相色谱法观察学习记忆过程中DG区细胞外液中谷氨酸(Glu)含量的动态变化;采用蛋白免疫印迹法检测海马组织和神经元细胞中c-Notch1、Notch1、p-CREB、BDNF等相关信号蛋白的表达情况。结果: 动物实验显示,与Control组相比,CatK Ⅱ组空间学习记忆能力明显减弱,并伴随着海马组织的c-Notch1、p-Akt、p-CREB和BDNF蛋白表达明显下降(P＜ 0.05);Control组和CatK II组DG区Glu含量均随水迷宫训练天数的增加而明显升高,但CatK Ⅱ组升高幅度均明显弱于Control组(P＜0.05)。细胞实验结果显示,与CON组和NC组相比,siCatK组CatK、c-Notch1、p-CREB和BDNF蛋白表达明显减少(P＜0.05)。结论: CatK可通过激活海马Notch1及其记忆相关信号蛋白,影响大鼠空间学习记忆功能。

关键词: 组织蛋白酶K, Notch1, 学习和记忆, 海马, 大鼠, 细胞培养

Abstract: Objective: To investigate the effects of Cathepsin K(CatK) on spatial learning and memory in rat hippocampus and its mechanisms. Methods: Twenty male SD rats were randomly divided into Control group and CatK inhibitor group(CatKⅡ group), which were microinjected with Cathepsin K specific inhibitor(0.5 μg/μl) and artificial cerebrospinal fluid in hippocampal DG area respectively with 5 days. The cultured hippocampal neuron cells were divided into control group (CON group), negative control group(NC group), siRNA interference group(siCatK group). Three re-wells were set for each group, and samples were collected 18～20 h after siRNA transfection. Morris water maze was used to evaluate spatial learning and memory function of rats. Meanwhile, dynamic changes of glutamate(Glu) content in extracellular fluid of DG region during learning and memory were observed by microdialysis and high performance liquid chromatography in conscious rats. Western blot was used to detect CatK-mediated Notch1 activation and other signal molecules. Results: Animal experiments showed that compared with the control group, the spatial learning and memory ability were decreased significantly in CatKII group, and the hippocampus protein expressions of c-Notch1, p-Akt, p-CREB and BDNF were also decreased significantly(P＜0.05); the levels of Glu in DG area of control group and CatK II group were increased significantly with Morris water maze training days, but the increase of CatK II group was significantly weaker than that of control group(P＜ 0.05). The results of cell experiment showed that the expressions of CatK, c-Notch1, p-CREB and BDNF in siCatK group were significantly lower than other groups (P＜0.05). Conclusion: CatK can affect the spatial learning and memory function of rats by activating Notch1 and its memory related signal protein in hippocampus.

Key words: Cathepsin K, Notch1, learning and memory, hippocampus, rat, cell culture

中图分类号:

R338

陈丽, 陈金信, 王心怡, 李香兰, 姜海英. 组织蛋白酶K对大鼠空间学习记忆功能的影响^*[J]. 中国应用生理学杂志, 2022, 38(5): 412-417.

CHEN Li, CHEN Jin-xin, WANG Xin-yi, LI Xiang-lan, JIANG Hai-ying. Effects of Cathepsin K on spatial learning and memory in rats[J]. CJAP, 2022, 38(5): 412-417.

参考文献

[1] Dai R, Wu Z, Chu HY, et al. Cathepsin K: The action in and beyond bone[J]. Front Cell Dev Biol, 2020, 8: 433.
[2] Hsu A, Podvin S, Hook V. Lysosomal cathepsin protease gene expression profiles in the human brain during normal development[J]. J Mol Neurosci, 2018, 65(4): 420-431.
[3] Dauth S, Sirbulescu RF, Jordans S, et al. Cathepsin K deficiency in mice induces structural and metabolic changes in the central nervous system that are associated with learning and memory deficits[J]. BMC Neurosci, 2011, 12: 74.
[4] Liu S, Wang Y, Worley PF, et al. The canonical Notch pathway effector RBP-J regulates neuronal plasticity and expression of GABA transporters in hippocampalnetworks[J]. Hippocampus, 2015, 25(5): 670-678.
[5] Vidak E, Javorsek U, Vizovisek M, et al. Cysteine cathepsins and their extracellular roles: Shaping the microenvironment[J]. Cells, 2019, 8(3): 264.
[6] Jiang H, Cheng XW, Shi GP, et al. Cathepsin K-mediated Notch1 activation contributes to neovascularization in response to hypoxia[J]. Nat Commun, 2014, 5: 3838.
[7] Wang S, Pan DX, Wang D, et al. Nitric oxide facilitatesactive avoidance learning via enhancement of glutamate levels in thehippocampal dentate gyrus[J]. Behav Brain Res, 2014, 271: 177-183.
[8] Wang F, Wan P, Wang W, et al. Dopamine in the hippocampal dentate gyrus modulates spatial learning via D1-like receptors[J]. Brain Res Bull, 2019, 144: 101-107.
[9] Wan P, Wang S, Zhang Y, et al. Involvement of dopamine D1 receptors of the hippocampal dentate gyrus in spatial learning and memory deficits in a rat model of vascular dementia[J]. Pharmazie, 2014, 69(9): 709-710.
[10] Alberi L, Liu S, Wang Y, et al. Activity-induced Notch signaling in neurons requires Arc/Arg3.1 and is essential for synaptic plasticity in hippocampal networks[J]. Neuron, 2011, 69(3): 437-444
[11] Wang Y, Chan SL, Miele L, et al. Involvement of Notch signaling in hippocampal synaptic plasticity[J]. Proc NatlAcad Sci USA, 2004, 101(25): 9458-9462.
[12] Asai M, Yagishita S, Iwata N, et al. An alternative metabolic pathway of amyloid precursor protein C-terminal fragments via cathepsin B in a human neuroglioma model[J]. Faseb J, 2011, 25(10): 3720-3730.
[13] Rosa E, Fahnestock M. CREB expression mediates amyloid β-induced basal BDNF downregulation[J]. Neurobiol Aging, 2015, 36(8): 2406-2413.
[14] Bartolotti N, Segura L, Lazarov O. Diminished CRE-induced plasticity is linked to memory deficits in familial Alzheimer's disease mice[J]. J Alzheimers Dis, 2016, 50(2): 477-489.
[15] Brai E, Marathe S, Astori S, et al. Notch1 regulates hippocampal plasticity through interaction with the reelin pathway, glutamatergic transmission and CREB signaling. Front Cell Neurosci, 2015, 9: 447.
[16] 李清山, 杨威, 潘艳芳, 等. 脑源性神经营养因子拮抗β淀粉样蛋白所致大鼠在体海马长时程增强的伤害[J]. 中国应用生理学杂志, 2012, 28(5): 425-429.
[17] 董雪帆, 王婷, 李甜, 等. APPswe/PS1dE9转基因小鼠小脑内突触素及BDNF /Trk-B蛋白表达的变化[J]. 中国应用生理学杂志, 2017, 33(6): 488-492.
[18] He W, Tian X, Yuan B, et al. Rosuvastatin improves neurite extension in cortical neurons through the Notch 1/BDNF pathway[J]. Neurol Res, 2019, 41(7): 658-664.

组织蛋白酶K对大鼠空间学习记忆功能的影响^*

Effects of Cathepsin K on spatial learning and memory in rats

PDF (PC)

可视化

摘要/Abstract

引用本文

使用本文

参考文献

相关文章 15

编辑推荐

Metrics

本文评价

[1]	张凯, 马敬霞, 马作旺, 杨禹, 李广平. 慢性间歇低氧对SD大鼠心房电重构的影响^*[J]. 中国应用生理学杂志, 2022, 38(5): 385-391.
[2]	钟锦, 史晋朝, 张倩, 孟金凤, 张倩倩, 李建国. Bdnf基因过表达慢病毒载体的构建及表达^*[J]. 中国应用生理学杂志, 2022, 38(5): 397-400.
[3]	邓筱悦, 帅念念, 刘时彦, 侯立力, 田绍文. 脑内糖原磷酸化酶抑制剂对大鼠急性癫痫发作神经炎症及记忆减退的改善作用^*[J]. 中国应用生理学杂志, 2022, 38(5): 406-411.
[4]	尹佳怡, 杲修杰, 张鑫垚, 郑鹏芳, 李小芳, 王瑞, 崔博. 40 Hz声光刺激对创伤诱发大鼠焦虑样行为的干预作用^*[J]. 中国应用生理学杂志, 2022, 38(5): 418-423.
[5]	王勇, 赵伟, 姜正林, 陈振华, 张欢. miR-125b-5p对创伤性脑损伤诱发大鼠认知功能障碍的干预作用及其机制^*[J]. 中国应用生理学杂志, 2022, 38(5): 424-429.
[6]	郑刘晔, 奚用勇, 张晗, 蒋与刚. 大鼠海马神经细胞VDAC1基因shRNA慢病毒表达载体的构建及干扰效果评价^*[J]. 中国应用生理学杂志, 2022, 38(5): 430-433.
[7]	陈津子, 李旭艳, 刘东梅, 龙舒婷, 李永媚, 陈泓冰. 纳米ZnO对人肺上皮细胞BEAS-2B细胞增殖和凋亡的影响^*[J]. 中国应用生理学杂志, 2022, 38(5): 443-447.
[8]	黄彩云, 陈那娜, 周菲, 张红梅, 杨小荣. 杏仁核SIRT1对慢性束缚应激大鼠抑郁样行为的影响^*[J]. 中国应用生理学杂志, 2022, 38(5): 458-463.
[9]	苟小红, 贺学农, 蒋世双, 陶武, 何朝辉. 糖皮质激素受体激动剂对神经病理性疼痛大鼠痛敏感反应的干预作用及其机制^*[J]. 中国应用生理学杂志, 2022, 38(5): 470-474.
[10]	杨滢霞, 郭育芬, 黄红红, 王凌星. 丁苯酞对睡眠剥夺大鼠脑额叶HMGB1和RAGE表达的影响^*[J]. 中国应用生理学杂志, 2022, 38(5): 480-484.
[11]	李姣姣, 平政, 王子文, 汪一波, 史春梦, 曹雪滨. 线粒体靶向小分子IR-61改善大鼠力竭性运动心脏损伤的实验研究^*[J]. 中国应用生理学杂志, 2022, 38(5): 497-503.
[12]	郭佳, 李志东, 肖传实, 边云飞. PUMA对高糖诱导的大鼠H9C2心肌细胞凋亡的作用及机制^*[J]. 中国应用生理学杂志, 2022, 38(5): 504-509.
[13]	严惠, 赵虎, 李论. 1-磷酸鞘氨醇(S1P)对H9c2心肌细胞肥大反应的保护作用^*[J]. 中国应用生理学杂志, 2022, 38(5): 510-514.
[14]	褚智恒, 何施燕, 王瑜, 朱若婷, 顾益玲, 陈佳玉. 紫云英苷对未分化胃癌细胞凋亡的影响 ^*[J]. 中国应用生理学杂志, 2022, 38(5): 520-524.
[15]	尼嘎拉·艾克热木, 高瑞娟, 唐雪纯, 孔良靖元, 高逸璇, 袁佳月, 马克涛, 李丽, 司军强. 丙磺舒对血小板衍生生长因子-BB诱发大鼠肺动脉平滑肌细胞迁移和增殖的干预作用^*[J]. 中国应用生理学杂志, 2022, 38(5): 543-548.