The enzyme heme oxygenase is presently the focus of numerous scientific research
projects and is meanwhile considered a cytoprotective enzyme, making it extremely
important for medicine.
One goal of this thesis was to adapt the HO-Assay for measurement of HO activity in
tissues of the pig. For this purpose the Assay should be optimized, allowing a precise
and sensitive measurement of the enzyme activity with the lowest possible error rate.
While using a BR dilution series, it was detected, that not all of the BR, a sample
contains, can be extracted into the chloroform-phase, which is used for the
photometric measurement. This loss was due to the presence of protein, can be
quantified and can now be used to correct the obtained results. In addition, the effect
of the HO-inhibitor ZnIXpp has been proofed and shown that the formation of BR in
pigs, as described for the rat, is specific, and therefore the assay is suitable for
measuring the HO activity. For the measurements obtained, it did not matter, as the
tissue was previously diluted. In addition, it was confirmed that the BVR activity is not
a limiting factor for the HO-Assay. However, it was shown that the HO activity less
strongly increases with increasing concentrations of protein. This can be attributed to
the interaction of BR with the proteins and its reduced extractability.
So it must be taken out, either to work with constant amounts of protein in an assay,
or to calculate the effect, is caused by the amount of protein, by using a correction