ULTRASTRUCTURAL DEMONSTRATION OF NADPH OXIDASE ACTIVITY IN KUPFFER AND ENDOTHELIAL CELLS USING AN ISOLATED LIVER PERFUSION SYSTEM

COMPARISON BETWEEN ICR/D-STRAIN AND JCL: WISTAR-STRAIN RATS

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In order to cytochemically demonstrate the NADPH dependent H₂O₂ generation system in the sinusoidal lining cells of the liver, the authors applied the modified method of Briggs et al. (5) to an isolated liver perfusion system devised by Sugano et al. (21) . The rat liver was perfused with the reaction medium containing 40mg NADPH, 40mg CeCl₃ 7H₂O and 2mg glucose in 100ml of 0.1M tris-maleate buffer solution (pH7.4), continuously supplied with a gas mixture (95%O₂ and 5%CO₂) . With respect to the specificity of the reaction, the in situ localization, and the amount of reaction products, adequate results were obtained with this system. Thus, the activity in the sinusoidal lining cells was compared between livers of the Jcl: Wistar-and the ICR/d-strain rats affected with spontaneously occurring cataracts and dermatitis.
Both Kupffer and endothelial cells showed conspicuous reactivity. A specific and significant reaction product was observed, adherent to the plasma membrane, on the external cell surface, including that of invaginated portions or coated pits. Furthermore, it was also noted on the internal surface of phagocytic vacuoles or coated vesicles. Comparing the two strains of rat, a much higher reactivity was shown in the liver of ICR/d-rat, suggesting that a defensive activity may be enhanced in the sinusoidal area to compensate for constitutional defects such as immunological disorders inherent in the ICR/d-strain.