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Brian G. Harbrecht, MD; Elizabeth M. Wirant; Young-Myeong Kim, PhD; Timothy R. Billiar, MD

Arch Surg. 1996;131(12):1266-1272.

Abstract
Objective
To investigate the effects of glucagon on nitric oxide (NO) synthesis in cultured rat hepatocytes.

Setting
Laboratory.

Materials
Male Sprague-Dawley rats (weight, 200-250 g).

Interventions
Isolated rat hepatocytes were cultured with interleukin-1 to stimulate NO synthesis. Glucagon was added at increasing concentrations (from 10^–9 to 2x 10^–5 mol/L) at the time of interleukin-1 stimulation. Selected cultures were treated with the adenylate cyclase inhibitor, SQ 22 536 (from 10^–5 to 10^–3 mol/L).

Main Outcome Measures
Nitric oxide synthesis was assessed by measuring the concentrations of culture supernatant nitrite and nitrite plus nitrate, hepatocyte nitric oxide synthase-2 (NOS-2) messenger RNA (mRNA), and NOS-2 protein.

Results
Interleukin-1 stimulated hepatocyte NO synthesis, and this synthesis was inhibited by glucagon in a dose-dependent manner. Glucagon inhibited the accumulation of supernatant nitrite and the expression of NOS-2 mRNA and NOS-2 protein. SQ 22 536 restored glucagon-induced decreases in NO synthesis.

Conclusions
Glucagon inhibits NO synthesis in interleukin-1–stimulated hepatocytes in vitro. This inhibition seems to be mediated by glucagon-induced changes in cyclic adenosine monophosphate.

Arch Surg. 1996;131:1266-1272

Author Affiliations
From the Department of Surgery, University of Pittsburgh, Pittsburgh, Pa.

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