Medical Research Essay

Medical Research Essay

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LCN2 expression in normal liver. Healthy rat livers were analyzed for LCN2 expression by Western blot (Suppl. Fig. 1A), RT-PCR (Suppl. Fig. 1B) and immunohistochemistry (Suppl. Fig. 1C). Since LCN2 is naturally expressed in tissues prone to microorganism exposure [9, 29], we used samples from organs expressing abundant quantities of LCN2 (lung, kidney, heart and trachea) as positive controls. In this analysis we found that LCN2 expression in normal liver was relatively low but still detectable by RT-PCR and that in liver cells positive for LCN2 in immunohistochemistry were only bile duct epithelia.
Expression kinetics of LCN2 during HSC activation and transdifferentiation. Based on the importance of lipid-transporting proteins for HSC homeostasis and activation, we hypothesized that LCN2 expression would be regulated during HSC transdifferentiation, possibly contributing to the cascade of molecular events involved in HSC/MFB activation. For determination of LCN2 expression during HSC transdifferentiation in vitro, we analyzed protein extracts from HSC/MFB at various culture time points by Western blot (Fig. 1A). Interestingly, LCN2 expression was found in early HSC cultures, however decreasing during prolonged culturing and rarely detectable in fully transdifferentiated MFB. Ongoing transdifferentiation was confirmed by increased collagen type I expression. RT-PCR was applied to determine whether this expression kinetics is caused by decreased activity of the LCN2 gene or by translational regulation. We found that the LCN2 mRNA levels were compatible with the results obtained in Western blot suggesting that downregulation of LCN2 during transdifferentiation is transcriptionally controlled (Fig. 1B).
Repeated hepatic insult lead...

... middle of paper ...

...matory mediators that both induce NF-kB, but only IL-1b was able to significantly induce LCN2 expression in HepG2 cells. This IL-1b specificity may be caused by the requirement of the NF-kB-binding cofactor IkB-z for transcriptional activation of the LCN2 promoter [37].
In summary, we did demonstrate that LCN2 is induced rapidly and secreted into systemic circulation during acute and chronic liver injury, and we identified injured hepatocytes as the main source of LCN2 production. We proved the proinflammatory cytokine IL-1 as the major inducer of LCN2 through NFB activation. Our data strongly points to an important role of LCN2 in liver homeostasis and its involvement in hepatocyte injury responses and additionally, LCN2 levels not to be a direct indicator of liver fibrosis, but rather a marker of liver damage instead which may imply a certain diagnostic value.

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