Cytochrome P450 3 A expression in pigs livers after 24-hour preservation in Biolasol solution depending on the type of transgenesis.
BACKGROUND
An insufficient number of organs for transplantation shows the need for the development of new technologies. Xenotransplantation might be the answer.


OBJECTIVE
To determine if the type of transgenesis influences the level of CYP3A4, which takes an active part in xenobiotics metabolism in livers after 24-hour storage, depending on the kind of solution used for preservation.


METHODS
The experiment was carried out on 30 livers of Polish White Landrace divided into 5 groups depending on transgene type. The following human genes were transferred: α1,2-fucosyltransferase (groups I and II), α-galactosidase (III), combined α1,2-fucosyltransferase/α-galactosidase transgene (IV), and livers without modification (V). The livers were perfused and subsequently stored for 24 hours in Ringer's solution (group I) or Biolasol solution (II-V). Reperfusion/reflush was performed. CYP3A29 isomer concentration was analyzed in liver specimens collected twice: 30 minutes after perfusion and 30 minutes after reperfusion/reflush. Expression of mRNA CYP3A29 was marked using RT-PCR analysis and of protein CYP3A29 using Western blotting technique.


RESULTS
The most significant decrease in protein CYP3A29 expression after 24-hour preservation was observed in group I (55.88% decrease), while the least significant was observed in group IV (10.44% decrease). mRNA expression evaluation was similar: the most significant decrease was observed in group I (87.8% decrease) and the least significant in group III (4.6% decrease).


CONCLUSIONS
α1,2-Fcosyltransferase transgene seems to influence mRNA and protein CYP3A expression in case of liver grafting and preservation for transplantation. CYP3A expression was also influenced by the kind of preservation solution used.
Department of General, Vascular and Transplant Surgery, Medical University of Silesia, Katowice, Poland. Electronic address: gregorydr@wp.pl.
G
G
Budziński
Department of Histology and Embryology, Medical University of Silesia, Katowice, Poland.
A
A
Suszka-Świtek
Department of General, Vascular and Transplant Surgery, Medical University of Silesia, Katowice, Poland.
P
P
Roman
Department of General, Vascular and Transplant Surgery, Medical University of Silesia, Katowice, Poland.
A
A
Caban
Department of General, Vascular and Transplant Surgery, Medical University of Silesia, Katowice, Poland.
G
G
Oczkowicz
Department of Histology and Embryology, Medical University of Silesia, Katowice, Poland.
E
E
Czech
Department of General, Vascular and Transplant Surgery, Medical University of Silesia, Katowice, Poland.
A
A
Mały
Department of Histology and Embryology, Medical University of Silesia, Katowice, Poland.
R
R
Wiaderkiewicz
The National Research Institute of Animal Production, Kraków-Balice, Poland.
Z
Z
Smorąg
"Biochefa" Pharmaceutical Research Production Plant, Sosnowiec, Poland.
F
F
Ryszka
Department of General, Vascular and Transplant Surgery, Medical University of Silesia, Katowice, Poland.
L
L
Cierpka
D000818:Animals; D030801:Animals, Genetically Modified; D051544:Cytochrome P-450 CYP3A / Q000235:genetics* / Q000378:metabolism; D005647:Fucosyltransferases / Q000235:genetics*; D018014:Gene Transfer Techniques; D006801:Humans; D008099:Liver / Q000378:metabolism*; D016031:Liver Transplantation; D019188:Organ Preservation Solutions; D010477:Perfusion; D012333:RNA, Messenger / Q000378:metabolism*; D015424:Reperfusion; D034421:Sus scrofa; D013552:Swine; D014183:Transplantation, Heterologous; D000519:alpha-Galactosidase / Q000235:genetics*
D019188:Organ Preservation Solutions; D012333:RNA, Messenger; D051544:Cytochrome P-450 CYP3A; D005647:Fucosyltransferases; D000519:alpha-Galactosidase