[Hydrodynamics-based transfer of human apolipoprotein A-I gene into mice: study of factors involving an efficacy and duration of the transferred gene expression in animals' liver]. / Perenos gena apolipoproteina A-I cheloveka mysham metodom gidrodinamicheskikh in''ektsii: faktory, vliiaiushchie na éffektivnost' i prodolzhitel'nost' ékspressii gena v pecheni zhivotnogo.

Human apolipoprotein A-I gene (apoA-I) plasmid expression vectors were transferred into mice by hydrodynamic injections into tail vein. Two types of expression vectors were used. First one -pCMVcapoAI contains cDNA of apo A-I driven by human cytomegalovirus early gene promoter (CMV). Second one--pAlg contains genomic locus of intron-containing apo A-I under control of own extended 5'-regulatory region (APOAI). Hydrodynamic intravenous injections of both expression vectors led to appearance of human apo A-I mRNA in the liver and human Apo A-I protein in the serum of injected mice. Dynamics of human Apo A-I content in the serum of mice injected by pCMVcapoAI and pAlg were different. When pCMVcapoAI was used, maximal concentration of human Apo A-I protein in the mouse serum was detected one day after injection with following decline to zero level during next two weeks. Under the same conditions injections of pAlg led to maximal level of human Apo A-I concentration in the mouse serum (up to 20 mkg/ml in some animals) on the 5th-7th day of experiment with following graduate decline during several months (human Apo A-I concentration in the serum of oldest analyzed mouse (6 months after injection) was about 25% of its maximal level in the same animal). Levels of human Apo A-I concentration in the mouse serum were compatible after injections of both expression vectors, in spite of much more strong activity of CMV promoter in comparison with APOAI in cultured human hepatoma cells HepG2. We ascribe the revealed difference in dynamics of human Apo A-I expression to delay of apo A-I transcription from pAlg vector, that was confirmed by nested RT-PCR. Significant level and long-term persistence of human Apo A-I in the serum of mice injected by pAlg could be explained by properties of APOAI or (and) exon-intron structure of genomic apo A-I gene. To test the role of APOAI in long-term expression of human Apo A-I in the mice we performed hydrodynamic injections of plasmid vectors containing cDNA of reporter gene encoding luciferase driven by variants of APOAI. No long-term expression of luciferase was found in the livers of injected mice. Therefore, our data suggest the role of exon-intron structure in maintaining of efficient and long-term expression of transferred human apo A-I.

[Status of lipid metabolism in rabbits, delivered from females with moderate alimentary hypercholesterolemia]. / Sostoianie lipidnogo obmena u krolikov, rodivshikhsia ot samok s umerennoi alimentarnoi giperkholesteinemiei.

Offspring of rabbits fed with cholesterol diet during pregnancy and/or lactation was more sensitive to fat load versus the offspring of rabbits fed with standard diet. Such enhancement of hyperlipidemic reaction is predominantly due to the enhanced rate of blood cholesterol increase (i.e., secretion of cholesterol-containing lipoproteins) but not to the decreased rate of cholesterol elimination from the blood stream. This was confirmed by monitoring the fate of human low-density lipoproteins intravenously injected in rabbits. The data indicate certain alterations of cholesterol metabolism regulation in the offspring of rabbits with alimentary hypercholesterolemia. Hypercholesterolemia of pregnant and lactating rabbits is important for the formation of the lipid metabolism in their offspring because cholesterol levels in the serum of female rabbits positively correlate with the expression of hypercholesterolemic and hypertriglyceridemia reactions of their offspring in response to fat load and with kinetic parameters which characterize the increase in cholesterol and its elimination from the blood stream during fat load.