Protein kinase Cγ play roles in hypoxic preconditioning-induced neuroprotection and determination of its interacting proteins in penumbra of mice with ischemic stroke

doi:10.3969/j.issn.1006-7795.2013.05.013

Abstract

Abstract:

Objective To validate the role of conventional protein kinase C (cPKC)γ in hypoxic precondition (HPC)- induced neuroprotection against brain ischemic injury, and determine cPKCγ -specific signaling proteins in penumbra of mice following ischemic stroke by using cPKCγ inhibitor and HPC mouse model.Methods Healthy adult male BALB/c mice were randomly divided into five groups, including normoxic sham, normoxic ischemia (I), ischemia following DMSO treatment (DMSO+I), ischemia post HPC (HPC+I), and ischemia after cPKCγ inhibitor Go6983 and HPC pretreatments. Using HPC and middle cerebral artery occlusion (MCAO)-induced ischemic stroke mouse models, the cPKCγ membrane translocation (activation) levels, cerebral infraction volum and cPKCγ -interacting proteins in ischemic penumbra were determined by Western blot, TTC staining, cPKCγ immunoprecipitation, two-dimensional electrophoresis (2-DE) and mass spectrometry (MS), respectively. Results Compared with MCAO-induced ischemia (I) group, cPKCγ membrane translocation (activation) levels was increased significantly in ischemic core and penumbra of HPC+I group, but cPKCγ inhibitor Go6983 pretreatment could inhibit the increases of cPKCγ membrane translocation in ischemic core, penumbra and contralateral cortex of HPC+Go6983+I mice (P<0.05, n=6 per group). cPKCγ inhibitor Go6983 abolished HPC-induced reduction in infarction size of mice with ischemic stroke (P<0.05, n=6 per group). In addition, Go6983 could inhibit the interaction between cPKCγ and partial interacting proteins in ischemic penumbra of HPC+I mice, including superoxide dismutase [Cu-Zn], DJ-1, UMP-CMP kinase, adenylate kinase isoenzyme 1, ubiquitin carboxyl-terminal hydrolase isozyme L1 (UCHL1), interleukin-16 and heat shock protein 70 (HSP70). Conclusion cPKCγ participates HPC-induced neuroprotection though regulating its interaction with partial interacting proteins in ischemic penumbra of mice following ischemic stroke.

Key words: hypoxic preconditioning, ischemic stroke, protein kinase C, neuroprotection, proteomics

CLC Number:

R339.5

HAN Song, ZHANG Nan, YIN Yanling, ZHAO Li, LUO Yanlin, LI Junfa. Protein kinase Cγ play roles in hypoxic preconditioning-induced neuroprotection and determination of its interacting proteins in penumbra of mice with ischemic stroke[J]. Journal of Capital Medical University, 2013, 34(5): 698-703.

0
/ / Recommend

Add to citation manager EndNote|Reference Manager|ProCite|BibTeX|RefWorks

URL: https://journal03.magtech.org.cn/Jweb_sdykdxxb/EN/10.3969/j.issn.1006-7795.2013.05.013

https://journal03.magtech.org.cn/Jweb_sdykdxxb/EN/Y2013/V34/I5/698

References

[1] Go A S, Mozaffarian D, Roger V L, et al. Heart disease and stroke statistics-2013 update: a report from the American Heart Association [J]. Circulation,2013,127(1): e6-e245.
[2] Kaste M. Stroke: advances in thrombolysis [J]. Lancet Neurol,2013,12(1): 2-4.
[3] Zhang N, Yin Y, Han S, et al. Hypoxic preconditioning induced neuroprotection against cerebral ischemic injuries and its cPKCgamma-mediated molecular mechanism [J]. Neurochem Int,2011, 58(6): 684-692.
[4] Bu X, Zhang N, Yang X, et al. Proteomic analysis of cPKCbetaⅡ-interacting proteins involved in HPC-induced neuroprotection against cerebral ischemia of mice [J]. J Neurochem,2011, 117(2): 346-356.
[5] Saito N, Shirai Y. Protein kinase C gamma (PKC gamma): function of neuron specific isotype [J]. J Biochem,2002, 132(5): 683-687.
[6] Wang F, Wang Y, Geng X, et al. Neuroprotective effect of acute ethanol administration in a rat with transient cerebral ischemia [J]. Stroke,2012, 43(1): 205-210.
[7] Werner D F, Kumar S, Criswell H E, et al. PKCgamma is required for ethanol-induced increases in GABA(A) receptor alpha4 subunit expression in cultured cerebral cortical neurons [J]. J Neurochem,2011,116(4): 554-563.
[8] Rex E B, Rankin M L, Yang Y, et al. Identification of RanBP 9/10 as interacting partners for protein kinase C (PKC) gamma/delta and the D1 dopamine receptor: regulation of PKC-mediated receptor phosphorylation [J]. Mol Pharmacol, 2010, 78(1): 69-80.
[9] 刘英超,庞琦. 重体脉瘤蛋白组学的研究进展[J].中华肿瘤防治杂志,2010,17(19):1595-1599.
[10] Liu J, Hou J, Xia Z Y, et al. Recombinant PTD-Cu/Zn SOD attenuates hypoxia-reoxygenation injury in cardiomyocytes [J]. Free Radic Res,2013; 47(5): 386-393.
[11] Wang L C, Chang C P, Chio C C, et al. Hypobaric hypoxia preconditioning attenuates experimental heatstroke syndromes via preinduction of heat shock protein 70 [J]. Am J Med Sci,2012,344(5): 383-390.

[1]	Xu Jianke, Liu Ning, Han Miao, Zhang Hongyang, Han Xinsheng. Analysis of the relationship between non-high-density lipoprotein cholesterol and arterial plaque characteristic in a cohort of ischemic stroke patients [J]. Journal of Capital Medical University, 2023, 44(1): 49-53.
[2]	Qin Linhui, Li Ning, Yang Yu, Yang Yong, Ren Changhong. Limb remote ischemic perconditioning combined with remote ischemic postconditioning promotes neurogenesis in rat models of ischemic stroke [J]. Journal of Capital Medical University, 2023, 44(1): 54-61.
[3]	Li Xue, Fan Junfen, Wang Rongliang, Ma Qingfeng, Luo Yumin, Zhao Haiping. Epigenetic regulation of HDAC2 on TBC protein family members of neutrophils in patients with ischemic stroke [J]. Journal of Capital Medical University, 2023, 44(1): 62-71.
[4]	Jiang Fucheng, Huang Jyumei, Feng Yuexian, Zhong Hongliang, Jia Jianwen, Yang Hongchao, Liu He, Liu Yang. The evaluation of the efficacy and prognosis of conscious sedation in endovascular treatment in acute ischemic stroke [J]. Journal of Capital Medical University, 2023, 44(1): 72-77.
[5]	Li Sen, Guo Xuewen, Zhao Xin, Huang Anqi, Lin Jingge, Li Xingmao, Li Xiu, Du Yifeng, Zhao Yongmei, Bai Jie, Xia Zhangyong. Correlation between glycemic variability and neurological deterioration in patients with acute ischemic stroke and diabetes mellitus [J]. Journal of Capital Medical University, 2023, 44(1): 78-84.
[6]	Gu Qiang, Yu Xiaojun, Zhang Jin. Analysis of clinical characteristics and prognosis of the very old and young old patients with mild ischemic stroke [J]. Journal of Capital Medical University, 2022, 43(3): 474-479.
[7]	Zhao Wenbo, Ren Changhong, Li Sijie, Ma Hongrui, Ji Xunming. Research and application of a new technical system of hypoxic and ischemic conditioning for prevention and treatment of ischemic stroke——The Second Prize of 2020 National Science and Technology Progress [J]. Journal of Capital Medical University, 2022, 43(1): 1-5.
[8]	Wang Xuan, Ma Chongyang, Zhang Yawen, Cheng Hongfa, Zhang Qiuxia. A network pharmacology approach to uncover the molecular mechanisms of Houshiheisan for treatment of ischemic stroke [J]. Journal of Capital Medical University, 2021, 42(1): 43-52.
[9]	Li Ming, Li Sijie, Wu Chuanjie, Zhao Wenbo, Gu Chaoxiong, Ji Xunming. Focusing on clinical problems and promoting the transformation of scientific and technological achievements——Professor Ji Xunming [J]. Journal of Capital Medical University, 2020, 41(5): 748-751.
[10]	Shan Yi, Li Jing, Xu Xiaoyin. Functional and molecular imaging of ischemic stroke——Professor Lu Jie [J]. Journal of Capital Medical University, 2020, 41(5): 788-794.
[11]	Wu Jian, Gu Yakun, Liu Jia. Research progresses in neuroprotective effects of ischemic/hypoxia preconditioning [J]. Journal of Capital Medical University, 2020, 41(4): 664-670.
[12]	Ding Hui, Cai Yanning, Chen Biao. Ganoderma lucidum extract regulates autophagy induced by MPP⁺ in Neuro-2a cells [J]. Journal of Capital Medical University, 2019, 40(4): 596-601.
[13]	Guo Shuang, Li Qing, Wu Fang, Liu Jiabin, Li Yan, Du Xiangying, Lu Jie, Li Kuncheng. Application of dual-energy CT in differentiating hemorrhage from contrast medium extravasation after endovascualr treatment of acute ischemic stroke [J]. Journal of Capital Medical University, 2018, 39(2): 173-177.
[14]	Cui Xu, Ji Fang, Shu Luowa, Li Junfa, Pan Chuxiong. cPKCγ membrane translocation plays roles in Herkinorin postconditioning against ischemia/reperfusion injury in MCAO mice [J]. Journal of Capital Medical University, 2017, 38(3): 336-341.
[15]	Liu Quanle, Li Tianfu, Wang Baoguo. Neuroprotective mechanism of sodium valproate and its application [J]. Journal of Capital Medical University, 2017, 38(3): 365-371.