Pharmaceutical Research

, Volume 23, Issue 2, pp 291–302 | Cite as

Protein Kinase C Family Members as a Target for Regulation of Blood–Brain Barrier Na,K,2Cl-Cotransporter During In Vitro Stroke Conditions and Nicotine Exposure

  • Tianzhi Yang
  • Karen E. Roder
  • G. Jayarama Bhat
  • Thomas J. Thekkumkara
  • Thomas J. Abbruscato
Research Paper

Purpose

The aim of the study is to identify specific protein kinase C (PKC) isoforms involvement in K+ transport mediated at altered blood–brain barrier (BBB) response to stroke conditions with prior nicotine exposure, which provides ways to intervene pharmacologically in PKC-mediated molecular pathways that could lead to effective treatment for smoking stroke patients.

Methods

Changes in PKC isoform levels were studied in the cytosolic and membrane fractions of bovine brain microvessel endothelial cells subjected to stroke conditions as well as nicotine/cotinine exposure. Furthermore, abluminal Na,K,2Cl-cotransporter (NKCC) activity regulated by specific conventional PKC isoform activators and inhibitors was investigated using rubidium (86Rb) uptake studies.

Results

Membrane-bound PKCα, PKCβI, and PKCɛ levels were increased after 6 h hypoxia/aglycemia, and this was attenuated by 24-h nicotine/cotinine exposure. Interestingly, membrane-bound PKCγ protein level was decreased after 6 h hypoxia/aglycemia and increased by 24-h nicotine/cotinine exposure. 86Rb uptake studies showed that basolateral NKCC activity was down-regulated by both a conventional PKC inhibitor and specific inhibitors for PKCα, PKCβ, and PKCɛ and was up-regulated by an activator of conventional PKCs during 6-h hypoxia/aglycemia treatment.

Conclusion

Specific PKC inhibitors or activators might be designed to individualize stroke therapies and improve health outcome for smokers by rebalancing ion transport into and out of the brain.

Key Words

blood–brain barrier Na,K,2Cl-cotransporter nicotine protein kinase C stroke 

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Copyright information

© Springer Science + Business Media, Inc. 2006

Authors and Affiliations

  • Tianzhi Yang
    • 1
  • Karen E. Roder
    • 1
  • G. Jayarama Bhat
    • 1
  • Thomas J. Thekkumkara
    • 1
  • Thomas J. Abbruscato
    • 1
  1. 1.Department of Pharmaceutical Sciences, School of PharmacyTexas Tech University Health Sciences CenterAmarilloUSA

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