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Comparative evaluation of immunohistochemistry and real-time PCR for measuring proinflammatory cytokines gene expression in livers of rats treated with gold nanoparticles
- Khan, Haseeb A., Ibrahim, Khalid E., Khan, Ayaat, Alrokayan, Salman H., Alhomida, Abdullah S., Lee, Yong-kyu
- Experimental and toxicologic pathology 2016 v.68 no.7 pp. 381-390
- drugs, gene expression, half life, immunohistochemistry, inflammation, interleukin-1beta, interleukin-6, liver, messenger RNA, nanogold, proteins, quantitative polymerase chain reaction, rats, tumor necrosis factor-alpha
- Gold nanoparticles (GNPs) possess promising applications in targeted drug delivery and controlled release of a variety of chemical agents. However, the immunocompatibility of GNPs is poorly understood. After exposure, GNPs preferentially tend to accumulate is liver, where they induce an acute phase proinflammatory response. We therefore compared the two techniques, immunohistochemistry and real-time PCR for measuring the protein and mRNA expressions of IL-1β, IL-6 and TNF-α in liver of rats after intraperitoneal injections (5μg/animal) of 10 and 50nm diameter GNPs for 1 and 5days. The results showed that both 10nm and 50nm GNPs induced an acute phase expression of proinflammatory cytokines that receded on day 5. The proinflammatory response on day 1 was comparatively more severe with 50nm GNPs than 10nm GNPs. A comparative evaluation between immunostaining and real-time PCR showed that the latter technique is more sensitive as it could detect the cytokines mRNA expression in control samples as well. This could be partly attributed to the amplification strategy used in real-time PCR and partly to the variations in the half lives of cytokines mRNA and their resulting proteins.