Activation of Cytosolic Cathepsin B Activity in the Brain by Traumatic Brain Injury and Inhibition by the Neutral pH Selective Inhibitor Probe Z-Arg-Lys-AOMK

IF 4.1 3区 医学 Q2 BIOCHEMISTRY & MOLECULAR BIOLOGY
Sonia Podvin, Jazmin Florio, Brian Spencer, Michael Mante, Estefani Guzman, Carlos Arias, Charles Mosier, Von V. Phan, Michael C. Yoon, Jehad Almaliti, Anthony J. O’Donoghue, William H. Gerwick, Robert A. Rissman and Vivian Hook*, 
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引用次数: 0

Abstract

Cathepsin B has been shown to contribute to deficits in traumatic brain injury (TBI), an important risk factor for Alzheimer’s disease (AD). Cathepsin B is elevated in TBI and AD patients, as well as in animal models of these conditions. Knockout of the cathepsin B gene results in amelioration of TBI-induced motor dysfunction and improvement of AD memory deficit in mice. The mechanism of cathepsin B pathogenesis in these brain disorders has been hypothesized to involve its translocation to the cytosol from its normal lysosomal location. This study, therefore, evaluated brain cytosolic cathepsin B activity in the controlled cortical impact (CCI) mouse model of TBI. CCI-TBI resulted in motor deficits demonstrated by the rotarod assay, brain tissue lesions, and disorganization of the hippocampus. Significantly, CCI-TBI increased cytosolic cathepsin B activity in the brain cortex in the ipsilateral brain hemisphere that received the CCI-TBI injury, with a concomitant decrease in the lysosomal fraction. Cathepsin B activity was monitored using the substrate Z-Nle-Lys-Arg-AMC which specifically detects cathepsin B activity but not other cysteine proteases. The normal lysosomal distribution of cathepsin B was observed by its discrete localization in brain cortical cells. CCI-TBI resulted in a more diffuse cellular distribution of cathepsin B consistent with translocation to the cytosol. Further studies utilized the novel neutral pH-selective inhibitor, Z-Arg-Lys-AOMK, that specifically inhibits cathepsin B at neutral pH 7.2 of the cytosol but not at acidic pH 4.6 of lysosomes. Daily administration of Z-Arg-Lys-AOMK (ip), beginning 1 day before CCI-TBI, resulted in the reduction of the increased cytosolic cathepsin B activity induced by CCI-TBI. The inhibitor also reduced cathepsin B activities in homogenates of the brain cortex and hippocampus which were increased by CCI-TBI. Furthermore, the Z-Arg-Lys-AOMK inhibitor resulted in the reduction of motor function deficit resulting from CCI-TBI. These findings demonstrate the activation of cytosolic cathepsin B activity in CCI-TBI mouse brain injury.

创伤性脑损伤对脑内组织蛋白酶B活性的激活及中性pH选择性抑制剂探针Z-Arg-Lys-AOMK的抑制作用
组织蛋白酶B已被证明与创伤性脑损伤(TBI)缺陷有关,TBI是阿尔茨海默病(AD)的一个重要危险因素。组织蛋白酶B在TBI和AD患者以及这些疾病的动物模型中升高。敲除组织蛋白酶B基因可改善小鼠tbi诱导的运动功能障碍和AD记忆缺陷。组织蛋白酶B在这些脑部疾病中的发病机制已被假设涉及其从正常溶酶体位置转移到细胞质。因此,本研究评估了脑外伤小鼠控制皮质冲击(CCI)模型中脑细胞质组织蛋白酶B的活性。CCI-TBI导致运动缺陷,rotarod实验证明,脑组织病变,海马组织紊乱。值得注意的是,在接受CCI-TBI损伤的同侧大脑半球,CCI-TBI增加了大脑皮层的胞质组织蛋白酶B活性,并伴有溶酶体分数的降低。使用底物Z-Nle-Lys-Arg-AMC监测组织蛋白酶B的活性,该底物专门检测组织蛋白酶B的活性,而不是其他半胱氨酸蛋白酶。组织蛋白酶B在脑皮质细胞中的离散定位表明其溶酶体分布正常。CCI-TBI导致组织蛋白酶B更分散的细胞分布,这与向细胞质的易位一致。进一步的研究使用了新的中性pH选择性抑制剂Z-Arg-Lys-AOMK,它在细胞质的中性pH为7.2时特异性抑制组织蛋白酶B,而在溶酶体的酸性pH为4.6时特异性抑制组织蛋白酶B。从CCI-TBI前1天开始,每天给药Z-Arg-Lys-AOMK (ip),可降低CCI-TBI引起的细胞内组织蛋白酶B活性升高。该抑制剂还降低了脑皮质和海马匀浆中组织蛋白酶B的活性,而CCI-TBI增加了组织蛋白酶B的活性。此外,Z-Arg-Lys-AOMK抑制剂可减少CCI-TBI引起的运动功能障碍。这些发现证实了CCI-TBI小鼠脑损伤中胞质组织蛋白酶B活性的激活。
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来源期刊
ACS Chemical Neuroscience
ACS Chemical Neuroscience BIOCHEMISTRY & MOLECULAR BIOLOGY-CHEMISTRY, MEDICINAL
CiteScore
9.20
自引率
4.00%
发文量
323
审稿时长
1 months
期刊介绍: ACS Chemical Neuroscience publishes high-quality research articles and reviews that showcase chemical, quantitative biological, biophysical and bioengineering approaches to the understanding of the nervous system and to the development of new treatments for neurological disorders. Research in the journal focuses on aspects of chemical neurobiology and bio-neurochemistry such as the following: Neurotransmitters and receptors Neuropharmaceuticals and therapeutics Neural development—Plasticity, and degeneration Chemical, physical, and computational methods in neuroscience Neuronal diseases—basis, detection, and treatment Mechanism of aging, learning, memory and behavior Pain and sensory processing Neurotoxins Neuroscience-inspired bioengineering Development of methods in chemical neurobiology Neuroimaging agents and technologies Animal models for central nervous system diseases Behavioral research
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