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Journal of Peking University (Health Sciences) ›› 2022, Vol. 54 ›› Issue (3): 421-426. doi: 10.19723/j.issn.1671-167X.2022.03.005

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Inhibition connexin 43 by mimetic peptide Gap27 mediates protective effects on 6-hydroxydopamine induced Parkinson's disease mouse model

Hui-hui QUAN,Wei-xing XU,Yu-ze QI,Qing-ru LI,Hui ZHOU,Jing HUANG*()   

  1. Department of Occupational and Environmental Health, Peking University School of Public Health, Beijing 100191, China
  • Received:2020-08-31 Online:2022-06-18 Published:2022-06-14
  • Contact: Jing HUANG E-mail:jing_huang@bjmu.edu.cn
  • Supported by:
    the National Natural Science Foundation of China(21577004);the Beijing Natural Science Foundation of(7162104)

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Abstract:

Objective: To explore whether the using of mimetic peptide Gap27, a selective inhibitor of connexin 43 (Cx43), could block the death of dopamine neurons and influence the expression of Cx43 in 6-hydroxydopamine (6-OHDA)-induced Parkinson's disease mouse models. Methods: Eighteen C57BL/6 mice were randomly divided into control group, 6-OHDA group and 6-OHDA+Gap27 group, with 6 mice in each group. Bilateral substantia nigra stereotactic injection was performed. The control group was injected with ascorbate solution, 6-OHDA group was injected with 6-OHDA solution, and 6-OHDA+Gap27 group was injected with 6-OHDA and Gap27 mixed solution. Immuno-histochemical staining was used to detect the number of dopamine neurons, quantitative real-time polymerase chain reaction (qRT-PCR) was used to detect the expression of Cx43 messenger ribonucleic acid (mRNA), immuno-fluorescence staining was used to detect the distribution of Cx43 protein, the contents of Cx43 protein and Cx43 phosphorylation at serine 368 (Cx43-ps368) in mouse midbrain were detected by Western blot. Results: After injection of 6-OHDA, numerous dopamine neurons in substantia nigra died as Cx43 content increased, Cx43-ps368 content decreased. Mixing Gap27 while injecting 6-OHDA could reduce the number of death dopamine neurons and weaken the changes of Cx43 and Cx43-ps368 content caused by 6-OHDA. The number of tyrosine hydroxylase (TH) immunoreactive positive neurons in 6-OHDA group decreased to 27.7% ± 0.02% of the control group (P < 0.01); The number of TH immunoreactive positive neurons in 6-OHDA+Gap27 group was (1.64±0.16) times higher than that in 6-OHDA group (P < 0.05); The content of total Cx43 protein in 6-OHDA group was (1.44±0.07) times higher than that in 6-OHDA+Gap27 group (P < 0.05) while (1.68±0.07) times higher than that in control group (P < 0.01). In 6-OHDA group, the content of Cx43-ps368 protein and its proportion in total Cx43 protein were significantly lower than that in 6-OHDA+Gap27 group (P < 0.05). Conclusion: In 6-OHDA mouse models, mimetic peptide Gap27 played a protective role in reducing the damage to substantia nigra dopamine neurons, which was induced by 6-OHDA. The overexpression of Cx43 protein might have neurotoxicity to dopamine neuron. Meanwhile, decreasing Cx43 protein level and keeping Cx43-ps368 protein level may be the protective mechanisms of Gap27.

Key words: Connexin 43, 6-hydroxydopamine, Peptide Gap27, Parkinson's disease

CLC Number: 

  • R392.3

Figure 1

Variation of DA neuron number in substantia nigra A, TH immunoreactive neurons of control group (DAB staining, n=3); B, TH immunoreactive neurons of 6-OHDA group (DAB staining, n=3); C, TH immunoreactive neurons of 6-OHDA+Gap27 group (DAB staining, n=3); D, relative number of TH positive neurons in substantia nigra. * P < 0.05, # P < 0.01. TH, tyrosine hydroxylase; 6-OHDA, 6-hydroxydopamine."

Figure 2

Changes of Cx43 protein content, gene expression and distribution A, the differences of Cx43 and Cx43-ps368 protein content among groups (n=3); B, relative expression of Cx43 protein, Cx43-ps368 protein, and Cx43-ps368/Cx43; C, relative expression of Cx43 mRNA (n=3); D, immunofluorescence co-staining of astrocytes (green) and Cx43 protein (red) in substantia nigra (n=3); E, relative expression of Cx43 protein fluorescence intensity. * P < 0.05, # P < 0.01. 6-OHDA, 6-hydroxydopamine; DAPI, 4′, 6-diamidino-2-phenylindole."

1 Hayes MT . Parkinson's disease and Parkinsonism[J]. Am J Med, 2019, 132 (7): 802- 807.
doi: 10.1016/j.amjmed.2019.03.001
2 Luo H , Xiang Y , Qu X , et al. Apelin-13 suppresses neuroinflammation against cognitive deficit in a streptozotocin-induced rat mo-del of Alzheimer's disease through activation of BDNF-TrkB signaling pathway[J]. Front Pharmacol, 2019, 10, 395.
doi: 10.3389/fphar.2019.00395
3 苏晓梅, 张丹参. 星形胶质细胞与神经退行性疾病的相关性[J]. 中国药理学与毒理学杂志, 2019, 33 (10): 868- 869.
4 Phatnani H , Maniatis T . Astrocytes in neurodegenerative disease[J]. Cold Spring Harb Perspect Biol, 2015, 7 (6): e020628.
doi: 10.1101/cshperspect.a020628
5 Diaz EF , Labra VC , Alvear TF , et al. Connexin 43 hemichannels and pannexin-1 channels contribute to the α-synuclein-induced dysfunction and death of astrocytes[J]. Glia, 2019, 67 (8): 1598- 1619.
6 Orellana JA , Sáez PJ , Cortés-Campos C , et al. Glucose increases intracellular free Ca(2+) in tanycytes via ATP released through connexin 43 hemichannels[J]. Glia, 2012, 60 (1): 53- 68.
doi: 10.1002/glia.21246
7 Leithe E , Mesnil M , Aasen T . The connexin 43 C-terminus: a tail of many tales[J]. Biochim Biophys Acta Biomembr, 2018, 1860 (1): 48- 64.
doi: 10.1016/j.bbamem.2017.05.008
8 Evans WH , Boitano S . Connexin mimetic peptides: specific inhi-bitors of gap-junctional intercellular communication[J]. Biochem Soc Trans, 2001, 29 (Pt 4): 606- 612.
9 Li X , Zhao H , Tan X , et al. Inhibition of connexin43 improves functional recovery after ischemic brain injury in neonatal rats[J]. Glia, 2015, 63 (9): 1553- 1567.
doi: 10.1002/glia.22826
10 Faniku C , O'Shaughnessy E , Lorraine C , et al. The connexin mimetic peptide gap27 and Cx43-knockdown reveal differential roles for connexin43 in wound closure events in skin model systems[J]. Int J Mol Sci, 2018, 9 (2): 604.
11 Evans WH , Leybaert L . Mimetic peptides as blockers of connexin channel-facilitated intercellular communication[J]. Cell Commun Adhes, 2007, 14 (6): 265- 273.
doi: 10.1080/15419060801891034
12 Solan JL , Lampe PD . Connexin43 phosphorylation: structural changes and biological effects[J]. Biochem J, 2009, 419 (2): 261- 272.
doi: 10.1042/BJ20082319
13 Ek-Vitorin JF , King TJ , Heyman NS , et al. Selectivity of conne-xin 43 channels is regulated through protein kinase C-dependent phosphorylation[J]. Circ Res, 2006, 98 (12): 1498- 1505.
doi: 10.1161/01.RES.0000227572.45891.2c
14 黄焕焕, 赵雨佳, 张慧峰, 等. 抑制星形胶质细胞Cx43可保护LPS诱导的多巴胺神经元损伤[J]. 毒理学杂志, 2018, 32 (3): 186- 194.
15 Wang ZY , Lian H , Zhou L , et al. Altered expression of d1 and d2 dopamine receptors in vagal neurons innervating the gastric muscularis externa in a Parkinson's disease rat model[J]. J Parkinsons Dis, 2016, 6 (2): 317- 323.
doi: 10.3233/JPD-160817
16 张蓓, 淫羊藿苷对6-OHDA诱导的帕金森病小鼠模型的保护作用及机制研究[D]. 贵州: 遵义医科大学, 2019.
17 Franklin-Keith BJ , Paxinos G . The mouse brain in stereotaxic coordinates[M]. 3th ed New York: Academic Press, 2007: 360
18 Cotter ML , Boitano S , Lampe PD , et al. The lipidated connexin mimetic peptide SRPTEKT-Hdc is a potent inhibitor of Cx43 channels with specificity for the pS368 phospho-isoform[J]. Am J Physiol, 2019, 317 (4): C825- C842.
doi: 10.1152/ajpcell.00160.2019
19 Rufer M , Wirth SB , Hofer A , et al. Regulation of connexin-43, GFAP, and FGF-2 is not accompanied by changes in astroglial coupling in MPTP-lesioned, FGF-2-treated Parkinsonian mice[J]. J Neurosci Res, 1996, 46 (5): 606- 617.
doi: 10.1002/(SICI)1097-4547(19961201)46:5<606::AID-JNR9>3.0.CO;2-N
20 Kawasaki A , Hayashi T , Nakachi K , et al. Modulation of conne-xin 43 in rotenone-induced model of Parkinson's disease[J]. Neuroscience, 2009, 160 (1): 61- 68.
doi: 10.1016/j.neuroscience.2009.01.080
21 Wang Y , Wu Z , Liu X , et al. Gastrodin ameliorates Parkinson's disease by downregulating connexin 43[J]. Mol Med Rep, 2013, 8 (2): 585- 590.
doi: 10.3892/mmr.2013.1535
22 韩雪洁, 哈力达·巴合提汗, 高华, 等. 黄芩苷可影响帕金森病模型大鼠纹状体星形胶质细胞缝隙连接蛋白43的表达[J]. 中国组织工程研究, 2018, 22 (16): 2542- 2548.
doi: 10.3969/j.issn.2095-4344.0222
23 Epifantseva I , Shaw RM . Intracellular trafficking pathways of Cx43 gap junction channels[J]. Biochim Biophys Acta Biomembr, 2018, 1860 (1): 40- 47.
doi: 10.1016/j.bbamem.2017.05.018
24 Charron G , Doudnikoff E , Canron MH , et al. Astrocytosis in Parkinsonism: considering tripartite striatal synapses in physiopathology?[J]. Front Aging Neurosci, 2014, 6, 1- 12.
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