Alteration of Protein Kinase C Isoforms in the Liver of Septic Rat

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Abstract
The present study investigated the alteration of protein kinase C (PKC) isoforms in rat liver during the progression of sepsis. Cecal ligation and puncture (CLP) model of polymicrobial sepsis was used, with early and late sepsis referring to those animals sacrificed at 9 and 18 h, respectively, after CLP. The protein contents of various PKC isoforms were quantified by Western blot and densitometric analysis. PKCα activity was performed after immunoprecipitation and assayed based on the incorporation rate of 32p from [γ-32p] adenosine triphosphate (ATP) into histone. The distribution of PKCα was evaluated by immunohistochemical staining. The steady state expression of PKCα mRNA was estimated by reverse transcriptase-polymerase chain reaction (RT-PCR). The results indicated that 1) five isoforms (α, β, δ, ε, ζ) could be detected in normal rat liver. PKCα and β were predominantly present in the cytosolic fraction, while membrane-associated PKCδ was more prominent than that of cytosolic fraction; 2) the protein content of membrane-associated PKCα was significantly decreased at early (P P