Abstract
The present study provides supportive evidence for the effective prevention and treatment of lipopolysaccharide (LPS)-induced hepatocyte injury in neonatal mice by N-acetylcysteine (NAC). Hepatocytes of neonatal mice were obtained through collagenase digestion of the liver. The hepatocytes were treated either with LPS (10 μg/mL) alone or with NAC (5 mmol/L) for 1 h before the addition of LPS (10 μg/mL). After LPS treatment, 12 wells of the cultured hepatocytes and supernatants were harvested at 0, 6, and 12 h, respectively. Levels of alanine aminotransferase (ALT) and nitric oxide (NO) in the supernatant were biochemically quantified and reverse transcription-polymerase chain reaction (RT-PCR) was performed to detect the expression of inducible nitric oxide synthase (iNOS) mRNA after different treatments. At 0 h following the treatment of primary cultured hepatocytes with LPS, the levels of ALT, NO and iNOS mRNA in the supernatant were (21.1 ± 4.78) u/L, (1.6 ± 0.31) μmol/L and 0.17 ± 0.023, respectively; at 6 h, (59.8 ± 8.59) u/L, (6.6 ± 0.81) μmol/L, and 0.71 ± 0.091; and at 12 h, (89.6 ± 15.30) u/L, (7.8 ± 1.01) μmol/L, and 0.71 ± 0.097. The levels of ALT, NO and iNOS mRNA at 6 and 12 h increased significantly, compared to those at 0 h (P < 0.01). In contrast to LPS treatment alone, pretreatment with NAC before LPS addition significantly reduced the levels of ALT, NO and iNOS mRNA in the supernatant at 6 h to (40.8 ± 7.30) u/L, (3.2 ± 0.71) μmol/L, and 0.41 ± 0.060; and at 12 h to (55.4 ± 5.48) u/L, (4.0 ± 0.71) μmol/L, and 0.40 ± 0.067, respectively (P < 0.01). However, the levels of ALT, NO and iNOS mRNA at 0 h did not change significantly with both treatment approaches. NAC has protective effects in hepatocytes of neonatal mice against LPS-induced injury as shown by the reduced levels of ALT, NO and iNOS mRNA when primary hepatocytes were treated with NAC prior to LPS stimulation. We postulate that NAC exhibits its protective function by inhibiting LPS-induced transcription of iNOS, resulting in decreased levels of NO.
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References
Li J, Quan N, Bray T M. Supplementation of N-acetylcysteine normalizes lipopolysaccharide induced nuclear factor kappaB activation and proinflammatory cytokine production during early rehabilitation of protein malnourished mice. J Nutr, 2002, 132(11): 3286–3292
Majano P L, Medina J, Zubía I, Sunyer L, Lara-Pezzi E, Maldonado-Rodríguez A, López-Cabrera M, Moreno-Otero R. N-Acetyl-cysteine modulates inducible nitric oxide synthase gene expression in human hepatocytes. J Hepatol, 2004, 40(4): 632–637
Weiskirchen R, Gressner A M. Isolation and culture of hepatic stellate cells. Methods Mol Med. 2005, 117: 99–113
Arterburn L M, Zurlo J, Yager J D, Overton R M, Heifetz A H. A morphological study of differentiated hepatocytes in vitro. Hepatology. 1995, 22(1): 175–187
Beath S V. Hepatic function and physiology in the newborn. Semin Neonatol, 2003, 8(5): 337–346
New K J, Eaton S, Elliott K R, Spitz L, Quant P A. Effect of lipopolysaccharide and cytokines on oxidative metabolism in neonatal rat hepatocytes. J Pediatr Surg, 2001, 36(2): 338–340
Bergamini S, Rota C, Canali R, Staffieri M, Daneri F, Bini A, Giovannini F, Tomasi A, Iannone A. N-acetylcysteine inhibits in vivo nitric oxide production by inducible nitric oxide synthase. Nitric Oxide, 2001, 5(4): 349–360
Zhang W Z, Cui S C, Wang J Q, Lang Z W, Zhu R P. Distribution of inducible nitric oxide synthase in the liver of severe hepatitis. Zhonghua Chuan Ran Bing Za Zhi, 2005, 23(1): 38–39 (in Chinese)
Labib R, Abdel-Rahman M S, Turkall R. N-acetylcysteine pretreatment decreases cocaine and endotoxin-induced hepatotoxicity. J Toxicol Environ Health A, 2003, 66(3): 223–239
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Translated from Chinese Journal of Pediatrics, 2007, 45(1): 30–33 [译自: 中华儿科杂志]
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Wang, L., Xu, J., Tian, Y. et al. Protective effect of N-acetylcysteine against lipopolysaccharide injury in hepatocytes of neonatal mice. Front. Med. China 2, 182–185 (2008). https://doi.org/10.1007/s11684-008-0034-x
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DOI: https://doi.org/10.1007/s11684-008-0034-x