Journal of Pure and Applied MicrobiologyVol. 6 No. 1

The Effect of Extracted Bacterial Salmonella enteritidis Lipopolysaccharide on Inducible Nitric Oxide Synthase in Human Liver Hepatocellular Carcinoma Cell Lines in Induction and Inhibition Conditions

Hossein Rastegar¹, Akram Mirzaei², Hamid Reza Ahmadi Ashtiani3,4,5, Mehdi Hedayati6 and Shahin Ahmadi7*

¹Food and Drug Control Laboratory and Research Center, Tehran, Iran. ²Department of Biology, Science and Research Branch, Islamic Azad University, Tehran, Iran. ³Biochemistry and Nutrition Department of Zanjan Medical University, Zanjan, Iran. 4Islamic Azad University, Pharmaceutical Sciences Branch, Tehran, Iran. 5Islamic Azad University, Dental Sciences Branch, Tehran, Iran. 6Obesity Research Center, Research Institute for Endocrine Sciences, Shahid Beheshti University of Medical Sciences, Tehran, Iran. 7Department of Veterinary, Islamic Azad University, Tabriz, Iran.

Received on 10 May 2011 and accepted on 30 June 2011

 

ABSTRACT

Hepatocellular carcinoma is the most ordinary type of liver cancer and most commonly appears in a patient with chronic viral hepatitis and with cirrhosis. Lipopolysaccharide motivates the hepatocyte cells and increases the production inducible cytokine with the high production of nitric oxide and reactive oxygen species. In this study, the rate change iNOS searched by adding lipopolysaccharide and the effect of inhibiting this inflammatory enzyme by L-NAME and combined effect of this in HepG2 cancer cells was assessed. Salmonella enteritidis lipopolysaccharide is extracted by methanol-chloroform method and silver stained SDS-PAGE electrophoresis bands. Four treatment groups of Human liver hepatocellular carcinoma cell lines stimulated with 100ng/ml lipopolysaccharide, 500µM L-NAME as inhibitor and were incubated for 12 and 24 hours. Variables including both increase and decrease inflammatory factor, iNOS, was assayed. The obtained results showed that initial activity of iNOS in not stimulated in HepG2 cell, 0.902U/ml after stimulation with lipopolysaccharide for 12 and 24hours (0.491U/ml) and (0.433U/ml) decreased and the effect of this inhibitor was also studied. The data showed that reducing iNOS in Human liver hepatocellular carcinoma cell lines correlated with the cell density and duration of incubation with LPS. Inhibition of enzyme associated with inflammation, with inhibitor substances; L-NAME, observed as well as lipopolysaccharide. We can potentially design drugs to treat a variety disease and cancer to use.

Keywords : Inducible Nitric Oxide Synthase, HepG2 Cell, Salmonella enteritidis- Lipopolysaccharide- SDS-PAGE.