Hypoxic Postconditioning Protects Astrocytes from Hypoxia/Reoxygenation Injury Through Decreasing Gap Junction Function

Yuchen Gu; Yiming Sun; Zhe Liu; Xuhui Tong; Shuying Dong

doi:doi:10.11648/j.ijcems.20220801.12

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Hypoxic Postconditioning Protects Astrocytes from Hypoxia/Reoxygenation Injury Through Decreasing Gap Junction Function

Yuchen Gu, Yiming Sun, Zhe Liu, Xuhui Tong, Shuying Dong

Published in International Journal of Clinical and Experimental Medical Sciences (Volume 8, Issue 1)

Received: 22 December 2021 Accepted: 11 January 2022 Published: 20 January 2022

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Abstract

Ischemic postconditioning was induced by brief cycles of ischemia/reperfusion (I/R) at the end of ischemia scavenge brain tissues from I/R injury in several animal models. However, the relationship between ischemic postconditioning and gap junction (GJ) yet to be explored. Here, we investigated whether the beneficial effect of hypoxic postconditioning involves in decrease of GJ function via establishing the hypoxia/reoxygenation (H/R) model with astrocytes to mimic the cerebral I/R. The primary astrocytes were exposed to 8 h hypoxia/24 h reoxygenation. Hypoxic postconditioning (HPC) was induced by 3 cycles of 10 min reoxygenation/10 min hypoxia after 8 h hypoxia. Before H/R, the retinoid acid was added for 24 h, and oleamide was applied for 1 h. Parachute dye coupling assay was used to evaluate GJ function. The viability and apoptosis of astrocytes was detected by MTT, flow cytometry and Hoechst 33258 staining, respectively. Finally, the Protein expression of Cx43, Bcl-2 and Bax was tested by western blotting, while the effect of Cx43-siRNA to H/R injury and HPC was explored by Cx43-siRNA transfection. It was found that HPC attenuated the expected increase in GJ function during reperfusion increased astrocyte viability and inhibit apoptosis. Compared with H/R group, the HPC group exhibit an increased expression of Cx43 and Bcl-2 protein, but decrease in Bax. Moreover, the pretreatment with retinoid strengthened the effect of ischemic/hypoxic postconditioning, while oleamide weakened it. We attributed these effects to the inhibited gap junctional intercellular communication (GJIC) induced by HPC through inhibition of Cx43 expression on cell surface, indicating that HPC protects astrocytes from H/R injury.

Published in	International Journal of Clinical and Experimental Medical Sciences (Volume 8, Issue 1)
DOI	10.11648/j.ijcems.20220801.12
Page(s)	9-19
Creative Commons	This is an Open Access article, distributed under the terms of the Creative Commons Attribution 4.0 International License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted use, distribution and reproduction in any medium or format, provided the original work is properly cited.
Copyright	Copyright © The Author(s), 2022. Published by Science Publishing Group

Keywords

Astrocyte, Gap Junction, Hypoxia/Reoxygenation, Hypoxic Postconditioning, Apoptosis

References

[1]	Benjamin, E. J., et al., Heart Disease and Stroke Statistics-2019 Update: A Report from the American Heart Association. Circulation, 2019. 139 (10): e56-e528. 10.1161/CIR.0000000000000659.
[2]	Chen, C., et al., The Roles of GABA in Ischemia-Reperfusion Injury in the Central Nervous System and Peripheral Organs. Oxid Med Cell Longev, 2019. 2019: 4028394. 10.1155/2019/4028394.
[3]	Yang, C. S., et al., Inhibition of cadmium-induced oxidative injury in rat primary astrocytes by the addition of antioxidants and the reduction of intracellular calcium. J Cell Biochem, 2008. 103 (3): 825-34. 10.1002/jcb.21452.
[4]	Hausenloy, D. J. and D. M. Yellon, The therapeutic potential of ischemic conditioning: an update. Nat Rev Cardiol, 2011. 8 (11): 619-29. 10.1038/nrcardio.2011.85.
[5]	Sewell, W. H., D. R. Koth, and C. E. Huggins, Ventricular fibrillation in dogs after sudden return of flow to the coronary artery. Surgery, 1955. 38 (6): 1050-3.
[6]	Grech, E. D. and D. R. Ramsdale, Termination of reperfusion arrhythmia by coronary artery occlusion. Br Heart J, 1994. 72 (1): 94-5. 10.1136/hrt.72.1.94.
[7]	Leconte, C., et al., Delayed hypoxic postconditioning protects against cerebral ischemia in the mouse. Stroke, 2009. 40 (10): 3349-55. 10.1161/STROKEAHA.109.557314.
[8]	Vetrovoy, O. V., E. A. Rybnikova, and M. O. Samoilov, Cerebral Mechanisms of Hypoxic/Ischemic Postconditioning. Biochemistry (Mosc), 2017. 82 (3): 392-400. 10.1134/S000629791703018X.
[9]	Mesnil, M., et al., Defective gap junctional intercellular communication in the carcinogenic process. BiochimBiophysActa, 2005. 1719 (1-2): 125-45. 10.1016/j.bbamem.2005.11.004.
[10]	He, B., et al., Tramadol and flurbiprofen depress the cytotoxicity of cisplatin via their effects on gap junctions. Clin Cancer Res, 2009. 15 (18): 5803-10. 10.1158/1078-0432.CCR-09-0811.
[11]	Shintani-Ishida, K., K. Unuma, and K. Yoshida, Ischemia enhances translocation of connexin43 and gap junction intercellular communication, thereby propagating contraction band necrosis after reperfusion. Circ J, 2009. 73 (9): 1661-8. 10.1253/circj.cj-09-0079.
[12]	Freitas-Andrade, M., et al., Danegaptide Enhances Astrocyte Gap Junctional Coupling and Reduces Ischemic Reperfusion Brain Injury in Mice. Biomolecules, 2020. 10 (3). 10.3390/biom10030353.
[13]	Tu, R. H., et al., Novel Functional Role of Heat Shock Protein 90 in Mitochondrial Connexin 43-Mediated Hypoxic Postconditioning. Cell PhysiolBiochem, 2017. 44 (3): 982-997. 10.1159/000485399.
[14]	Pagliaro, P., et al., Mitochondria in Cardiac Postconditioning. Front Physiol, 2018. 9: 287. 10.3389/fphys.2018.00287.
[15]	Wang, H. C., et al., Hypoxic postconditioning enhances the survival and inhibits apoptosis of cardiomyocytes following reoxygenation: role of peroxynitrite formation. Apoptosis, 2006. 11 (8): 1453-60. 10.1007/s10495-006-7786-z.
[16]	Chen, X. Q., et al., Lipoxin A4-induced heme oxygenase-1 protects cardiomyocytes against hypoxia/reoxygenation injury via p38 MAPK activation and Nrf2/ARE complex. PLoS One, 2013. 8 (6): e67120. 10.1371/journal.pone.0067120.
[17]	Chen, J., et al., A synthetic steroid 5alpha-androst-3beta, 5, 6beta-triol blocks hypoxia/reoxygenation-induced neuronal injuries via protection of mitochondrial function. Steroids, 2013. 78 (10): 996-1002. 10.1016/j.steroids.2013.06.004.
[18]	Guo, G. and N. R. Bhat, Hypoxia/reoxygenation differentially modulates NF-kappaB activation and iNOS expression in astrocytes and microglia. Antioxid Redox Signal, 2006. 8 (5-6): 911-8. 10.1089/ars.2006.8.911.
[19]	Mohammad, I. S., et al., Phytocosmeceutical formulation development, characterization and its in-vivo investigations. Biomed Pharmacother, 2018. 107: 806-817. 10.1016/j.biopha.2018.08.024.
[20]	Mohammad, I. S., W. He, and L. Yin, A Smart Paclitaxel-Disulfiram Nanococrystals for Efficient MDR Reversal and Enhanced Apoptosis. Pharm Res, 2018. 35 (4): 77. 10.1007/s11095-018-2370-0.
[21]	Goldberg, G. S., J. F. Bechberger, and C. C. Naus, A pre-loading method of evaluating gap junctional communication by fluorescent dye transfer. Biotechniques, 1995. 18 (3): 490-7.
[22]	Koreen, I. V., et al., Tetracycline-regulated expression enables purification and functional analysis of recombinant connexin channels from mammalian cells. Biochem J, 2004. 383 (Pt 1): 111-9. 10.1042/BJ20040806.
[23]	Wu, J. F., et al., Gefitinib enhances oxaliplatin-induced apoptosis mediated by Src and PKC-modulated gap junction function. Oncol Rep, 2016. 36 (6): 3251-3258. 10.3892/or.2016.5156.
[24]	Epifantseva, I. and R. M. Shaw, Intracellular trafficking pathways of Cx43 gap junction channels. Biochim Biophys Acta Biomembr, 2018. 1860 (1): 40-47. 10.1016/j.bbamem.2017.05.018.
[25]	Zhao, H., R. M. Sapolsky, and G. K. Steinberg, Interrupting reperfusion as a stroke therapy: ischemic postconditioning reduces infarct size after focal ischemia in rats. J Cereb Blood Flow Metab, 2006. 26 (9): 1114-21. 10.1038/sj.jcbfm.9600348.
[26]	Joo, S. P., et al., Ischemic postconditioning protects against focal cerebral ischemia by inhibiting brain inflammation while attenuating peripheral lymphopenia in mice. Neuroscience, 2013. 243: 149-57. 10.1016/j.neuroscience.2013.03.062.
[27]	Liang, J. M., et al., Role of mitochondrial function in the protective effects of ischaemicpostconditioning on ischaemia/reperfusion cerebral damage. J Int Med Res, 2013. 41 (3): 618-27. 10.1177/0300060513476587.
[28]	Martinou, J. C. and R. J. Youle, Mitochondria in apoptosis: Bcl-2 family members and mitochondrial dynamics. Dev Cell, 2011. 21 (1): 92-101. 10.1016/j.devcel.2011.06.017.
[29]	Dong, L., et al., Vitexin protects against myocardial ischemia/reperfusion injury in Langendorff-perfused rat hearts by attenuating inflammatory response and apoptosis. Food Chem Toxicol, 2011. 49 (12): 3211-6. 10.1016/j.fct.2011.09.040.
[30]	Li, T., et al., Overexpression of apoptosis inducing factor aggravates hypoxic-ischemic brain injury in neonatal mice. Cell Death Dis, 2020. 11 (1): 77. 10.1038/s41419-020-2280-z.
[31]	Swanton, E., et al., Bcl-2 regulates a caspase-3/caspase-2 apoptotic cascade in cytosolic extracts. Oncogene, 1999. 18 (10): 1781-7. 10.1038/sj.onc.1202490.
[32]	Namura, S., et al., Activation and cleavage of caspase-3 in apoptosis induced by experimental cerebral ischemia. J Neurosci, 1998. 18 (10): 3659-68.
[33]	Belousov, A. B. and J. D. Fontes, Neuronal gap junctions: making and breaking connections during development and injury. Trends Neurosci, 2013. 36 (4): 227-36. 10.1016/j.tins.2012.11.001.
[34]	Zhang, L., et al., Protective effects of carbenoxolone are associated with attenuation of oxidative stress in ischemic brain injury. Neurosci Bull, 2013. 29 (3): 311-20. 10.1007/s12264-013-1342-y.
[35]	Chew, S. S., et al., Role of connexin43 in central nervous system injury. ExpNeurol, 2010. 225 (2): 250-61. 10.1016/j.expneurol.2010.07.014.

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APA Style

Yuchen Gu, Yiming Sun, Zhe Liu, Xuhui Tong, Shuying Dong. (2022). Hypoxic Postconditioning Protects Astrocytes from Hypoxia/Reoxygenation Injury Through Decreasing Gap Junction Function. International Journal of Clinical and Experimental Medical Sciences, 8(1), 9-19. https://doi.org/10.11648/j.ijcems.20220801.12

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ACS Style

Yuchen Gu; Yiming Sun; Zhe Liu; Xuhui Tong; Shuying Dong. Hypoxic Postconditioning Protects Astrocytes from Hypoxia/Reoxygenation Injury Through Decreasing Gap Junction Function. Int. J. Clin. Exp. Med. Sci. 2022, 8(1), 9-19. doi: 10.11648/j.ijcems.20220801.12

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AMA Style

Yuchen Gu, Yiming Sun, Zhe Liu, Xuhui Tong, Shuying Dong. Hypoxic Postconditioning Protects Astrocytes from Hypoxia/Reoxygenation Injury Through Decreasing Gap Junction Function. Int J Clin Exp Med Sci. 2022;8(1):9-19. doi: 10.11648/j.ijcems.20220801.12

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@article{10.11648/j.ijcems.20220801.12,
  author = {Yuchen Gu and Yiming Sun and Zhe Liu and Xuhui Tong and Shuying Dong},
  title = {Hypoxic Postconditioning Protects Astrocytes from Hypoxia/Reoxygenation Injury Through Decreasing Gap Junction Function},
  journal = {International Journal of Clinical and Experimental Medical Sciences},
  volume = {8},
  number = {1},
  pages = {9-19},
  doi = {10.11648/j.ijcems.20220801.12},
  url = {https://doi.org/10.11648/j.ijcems.20220801.12},
  eprint = {https://article.sciencepublishinggroup.com/pdf/10.11648.j.ijcems.20220801.12},
  abstract = {Ischemic postconditioning was induced by brief cycles of ischemia/reperfusion (I/R) at the end of ischemia scavenge brain tissues from I/R injury in several animal models. However, the relationship between ischemic postconditioning and gap junction (GJ) yet to be explored. Here, we investigated whether the beneficial effect of hypoxic postconditioning involves in decrease of GJ function via establishing the hypoxia/reoxygenation (H/R) model with astrocytes to mimic the cerebral I/R. The primary astrocytes were exposed to 8 h hypoxia/24 h reoxygenation. Hypoxic postconditioning (HPC) was induced by 3 cycles of 10 min reoxygenation/10 min hypoxia after 8 h hypoxia. Before H/R, the retinoid acid was added for 24 h, and oleamide was applied for 1 h. Parachute dye coupling assay was used to evaluate GJ function. The viability and apoptosis of astrocytes was detected by MTT, flow cytometry and Hoechst 33258 staining, respectively. Finally, the Protein expression of Cx43, Bcl-2 and Bax was tested by western blotting, while the effect of Cx43-siRNA to H/R injury and HPC was explored by Cx43-siRNA transfection. It was found that HPC attenuated the expected increase in GJ function during reperfusion increased astrocyte viability and inhibit apoptosis. Compared with H/R group, the HPC group exhibit an increased expression of Cx43 and Bcl-2 protein, but decrease in Bax. Moreover, the pretreatment with retinoid strengthened the effect of ischemic/hypoxic postconditioning, while oleamide weakened it. We attributed these effects to the inhibited gap junctional intercellular communication (GJIC) induced by HPC through inhibition of Cx43 expression on cell surface, indicating that HPC protects astrocytes from H/R injury.},
 year = {2022}
}

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TY  - JOUR
T1  - Hypoxic Postconditioning Protects Astrocytes from Hypoxia/Reoxygenation Injury Through Decreasing Gap Junction Function
AU  - Yuchen Gu
AU  - Yiming Sun
AU  - Zhe Liu
AU  - Xuhui Tong
AU  - Shuying Dong
Y1  - 2022/01/20
PY  - 2022
N1  - https://doi.org/10.11648/j.ijcems.20220801.12
DO  - 10.11648/j.ijcems.20220801.12
T2  - International Journal of Clinical and Experimental Medical Sciences
JF  - International Journal of Clinical and Experimental Medical Sciences
JO  - International Journal of Clinical and Experimental Medical Sciences
SP  - 9
EP  - 19
PB  - Science Publishing Group
SN  - 2469-8032
UR  - https://doi.org/10.11648/j.ijcems.20220801.12
AB  - Ischemic postconditioning was induced by brief cycles of ischemia/reperfusion (I/R) at the end of ischemia scavenge brain tissues from I/R injury in several animal models. However, the relationship between ischemic postconditioning and gap junction (GJ) yet to be explored. Here, we investigated whether the beneficial effect of hypoxic postconditioning involves in decrease of GJ function via establishing the hypoxia/reoxygenation (H/R) model with astrocytes to mimic the cerebral I/R. The primary astrocytes were exposed to 8 h hypoxia/24 h reoxygenation. Hypoxic postconditioning (HPC) was induced by 3 cycles of 10 min reoxygenation/10 min hypoxia after 8 h hypoxia. Before H/R, the retinoid acid was added for 24 h, and oleamide was applied for 1 h. Parachute dye coupling assay was used to evaluate GJ function. The viability and apoptosis of astrocytes was detected by MTT, flow cytometry and Hoechst 33258 staining, respectively. Finally, the Protein expression of Cx43, Bcl-2 and Bax was tested by western blotting, while the effect of Cx43-siRNA to H/R injury and HPC was explored by Cx43-siRNA transfection. It was found that HPC attenuated the expected increase in GJ function during reperfusion increased astrocyte viability and inhibit apoptosis. Compared with H/R group, the HPC group exhibit an increased expression of Cx43 and Bcl-2 protein, but decrease in Bax. Moreover, the pretreatment with retinoid strengthened the effect of ischemic/hypoxic postconditioning, while oleamide weakened it. We attributed these effects to the inhibited gap junctional intercellular communication (GJIC) induced by HPC through inhibition of Cx43 expression on cell surface, indicating that HPC protects astrocytes from H/R injury.
VL  - 8
IS  - 1
ER  -

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Author Information

Yuchen Gu

Department of Pharmacy, the First Affiliated Hospital of Bengbu Medical College, Bengbu, China
Yiming Sun

Department of Pharmacy, the First Affiliated Hospital of Bengbu Medical College, Bengbu, China
Zhe Liu

Department of Pharmacy, the First Affiliated Hospital of Bengbu Medical College, Bengbu, China
Xuhui Tong

College of Pharmacy, Bengbu Medical College, Bengbu, China
Shuying Dong

College of Pharmacy, Bengbu Medical College, Bengbu, China

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APA Style

Yuchen Gu, Yiming Sun, Zhe Liu, Xuhui Tong, Shuying Dong. (2022). Hypoxic Postconditioning Protects Astrocytes from Hypoxia/Reoxygenation Injury Through Decreasing Gap Junction Function. International Journal of Clinical and Experimental Medical Sciences, 8(1), 9-19. https://doi.org/10.11648/j.ijcems.20220801.12

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ACS Style

Yuchen Gu; Yiming Sun; Zhe Liu; Xuhui Tong; Shuying Dong. Hypoxic Postconditioning Protects Astrocytes from Hypoxia/Reoxygenation Injury Through Decreasing Gap Junction Function. Int. J. Clin. Exp. Med. Sci. 2022, 8(1), 9-19. doi: 10.11648/j.ijcems.20220801.12

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AMA Style

Yuchen Gu, Yiming Sun, Zhe Liu, Xuhui Tong, Shuying Dong. Hypoxic Postconditioning Protects Astrocytes from Hypoxia/Reoxygenation Injury Through Decreasing Gap Junction Function. Int J Clin Exp Med Sci. 2022;8(1):9-19. doi: 10.11648/j.ijcems.20220801.12

Copy | Download

@article{10.11648/j.ijcems.20220801.12,
  author = {Yuchen Gu and Yiming Sun and Zhe Liu and Xuhui Tong and Shuying Dong},
  title = {Hypoxic Postconditioning Protects Astrocytes from Hypoxia/Reoxygenation Injury Through Decreasing Gap Junction Function},
  journal = {International Journal of Clinical and Experimental Medical Sciences},
  volume = {8},
  number = {1},
  pages = {9-19},
  doi = {10.11648/j.ijcems.20220801.12},
  url = {https://doi.org/10.11648/j.ijcems.20220801.12},
  eprint = {https://article.sciencepublishinggroup.com/pdf/10.11648.j.ijcems.20220801.12},
  abstract = {Ischemic postconditioning was induced by brief cycles of ischemia/reperfusion (I/R) at the end of ischemia scavenge brain tissues from I/R injury in several animal models. However, the relationship between ischemic postconditioning and gap junction (GJ) yet to be explored. Here, we investigated whether the beneficial effect of hypoxic postconditioning involves in decrease of GJ function via establishing the hypoxia/reoxygenation (H/R) model with astrocytes to mimic the cerebral I/R. The primary astrocytes were exposed to 8 h hypoxia/24 h reoxygenation. Hypoxic postconditioning (HPC) was induced by 3 cycles of 10 min reoxygenation/10 min hypoxia after 8 h hypoxia. Before H/R, the retinoid acid was added for 24 h, and oleamide was applied for 1 h. Parachute dye coupling assay was used to evaluate GJ function. The viability and apoptosis of astrocytes was detected by MTT, flow cytometry and Hoechst 33258 staining, respectively. Finally, the Protein expression of Cx43, Bcl-2 and Bax was tested by western blotting, while the effect of Cx43-siRNA to H/R injury and HPC was explored by Cx43-siRNA transfection. It was found that HPC attenuated the expected increase in GJ function during reperfusion increased astrocyte viability and inhibit apoptosis. Compared with H/R group, the HPC group exhibit an increased expression of Cx43 and Bcl-2 protein, but decrease in Bax. Moreover, the pretreatment with retinoid strengthened the effect of ischemic/hypoxic postconditioning, while oleamide weakened it. We attributed these effects to the inhibited gap junctional intercellular communication (GJIC) induced by HPC through inhibition of Cx43 expression on cell surface, indicating that HPC protects astrocytes from H/R injury.},
 year = {2022}
}

Copy | Download

TY  - JOUR
T1  - Hypoxic Postconditioning Protects Astrocytes from Hypoxia/Reoxygenation Injury Through Decreasing Gap Junction Function
AU  - Yuchen Gu
AU  - Yiming Sun
AU  - Zhe Liu
AU  - Xuhui Tong
AU  - Shuying Dong
Y1  - 2022/01/20
PY  - 2022
N1  - https://doi.org/10.11648/j.ijcems.20220801.12
DO  - 10.11648/j.ijcems.20220801.12
T2  - International Journal of Clinical and Experimental Medical Sciences
JF  - International Journal of Clinical and Experimental Medical Sciences
JO  - International Journal of Clinical and Experimental Medical Sciences
SP  - 9
EP  - 19
PB  - Science Publishing Group
SN  - 2469-8032
UR  - https://doi.org/10.11648/j.ijcems.20220801.12
AB  - Ischemic postconditioning was induced by brief cycles of ischemia/reperfusion (I/R) at the end of ischemia scavenge brain tissues from I/R injury in several animal models. However, the relationship between ischemic postconditioning and gap junction (GJ) yet to be explored. Here, we investigated whether the beneficial effect of hypoxic postconditioning involves in decrease of GJ function via establishing the hypoxia/reoxygenation (H/R) model with astrocytes to mimic the cerebral I/R. The primary astrocytes were exposed to 8 h hypoxia/24 h reoxygenation. Hypoxic postconditioning (HPC) was induced by 3 cycles of 10 min reoxygenation/10 min hypoxia after 8 h hypoxia. Before H/R, the retinoid acid was added for 24 h, and oleamide was applied for 1 h. Parachute dye coupling assay was used to evaluate GJ function. The viability and apoptosis of astrocytes was detected by MTT, flow cytometry and Hoechst 33258 staining, respectively. Finally, the Protein expression of Cx43, Bcl-2 and Bax was tested by western blotting, while the effect of Cx43-siRNA to H/R injury and HPC was explored by Cx43-siRNA transfection. It was found that HPC attenuated the expected increase in GJ function during reperfusion increased astrocyte viability and inhibit apoptosis. Compared with H/R group, the HPC group exhibit an increased expression of Cx43 and Bcl-2 protein, but decrease in Bax. Moreover, the pretreatment with retinoid strengthened the effect of ischemic/hypoxic postconditioning, while oleamide weakened it. We attributed these effects to the inhibited gap junctional intercellular communication (GJIC) induced by HPC through inhibition of Cx43 expression on cell surface, indicating that HPC protects astrocytes from H/R injury.
VL  - 8
IS  - 1
ER  -

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