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N-3 polyunsaturated fatty acids attenuates triglyceride and inflammatory factors level in hfat-1 transgenic pigs.

Liu X, Pang D, Yuan T, Li Z, Li Z, Zhang M, Ren W, Ouyang H, Tang X - Lipids Health Dis (2016)

Bottom Line: Although beneficial effects of n-3 PUFAs have been observed in a number of studies, the mechanisms involved in these effects have yet to be discovered.Gas chromatography results demonstrated that the total n-3 PUFAs in the ear tissues of the transgenic founders were 2-fold higher than the wild-type pigs.The basal levels of the inflammation mediators tumor necrosis factor-α (TNF-α), monocyte chemoattractant protein-1 (MCP-1) and interleukin-6 (IL-6) in transgenic pigs were inhibited markedly compared with the wild-type pigs.

View Article: PubMed Central - PubMed

Affiliation: Jilin Provincial Model Animal Engineering Research Center, College of Animal Sciences, Jilin University, Xi'an Road, 5333#, Jilin, 130062, China.

ABSTRACT

Background: The consumption of n-3 polyunsaturated fatty acids (PUFAs) is important to human health, especially in cases of cardiovascular disease. Although beneficial effects of n-3 PUFAs have been observed in a number of studies, the mechanisms involved in these effects have yet to be discovered.

Methods: We generated hfat-1 transgenic pigs with traditional somatic cell nuclear transfer (SCNT) technology. The fatty acid composition in ear tissue of pigs were detected with gas chromatography. The cholesterol, triglycerides (TAG) and inflammation mediators in circulation were investigated.

Results: The hfat-1 transgenic pigs were developed which accumulate high levels of n-3 PUFAs than wild-types pigs. Gas chromatography results demonstrated that the total n-3 PUFAs in the ear tissues of the transgenic founders were 2-fold higher than the wild-type pigs. A lipid analysis demonstrated that the levels of TAG in the transgenic pigs were decreased significantly. The basal levels of the inflammation mediators tumor necrosis factor-α (TNF-α), monocyte chemoattractant protein-1 (MCP-1) and interleukin-6 (IL-6) in transgenic pigs were inhibited markedly compared with the wild-type pigs.

Conclusions: These results suggest that n-3 PUFAs accumulation in vivo may have beneficial effects on vascular and hfat-1 transgenic pigs may be a useful tool for investigating the involved mechanisms.

No MeSH data available.


Related in: MedlinePlus

Identification of G418-resistant clones and hfat-1 transgenic pigs. Analyse of the hfat-1 gene and neomycin gene integration into the pig genome in the G418-resistant clones were performed using a PCR assay with the primer pairs of hfat-1 identi (458 bp) (a) and neo identi (790 bp) (b). a 1–5, five G418-resistant clones, on which SCNT was individually performed in the following procedure. 6, hfat-1 plasmid as a positive control. 7–8, negative controls. b 1, hfat-1 plasmid as a positive control. 2–3, negative controls. 4–8, five G418-resistant clones. The hfat-1 gene integration into the pig genome and transcription in the transgenic pigs were analyzed using PCR (c) and RT-PCR (d), respectively, with the hfat-1 identi primer pair (458 bp). c 1–5, five hfat-1 transgenic pigs. 6, hfat-1 plasmid as a positive control. 7–8, two wild-type pigs. d 1–5, five hfat-1 transgenic pigs. 6–7, two wild-type pigs
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Fig2: Identification of G418-resistant clones and hfat-1 transgenic pigs. Analyse of the hfat-1 gene and neomycin gene integration into the pig genome in the G418-resistant clones were performed using a PCR assay with the primer pairs of hfat-1 identi (458 bp) (a) and neo identi (790 bp) (b). a 1–5, five G418-resistant clones, on which SCNT was individually performed in the following procedure. 6, hfat-1 plasmid as a positive control. 7–8, negative controls. b 1, hfat-1 plasmid as a positive control. 2–3, negative controls. 4–8, five G418-resistant clones. The hfat-1 gene integration into the pig genome and transcription in the transgenic pigs were analyzed using PCR (c) and RT-PCR (d), respectively, with the hfat-1 identi primer pair (458 bp). c 1–5, five hfat-1 transgenic pigs. 6, hfat-1 plasmid as a positive control. 7–8, two wild-type pigs. d 1–5, five hfat-1 transgenic pigs. 6–7, two wild-type pigs

Mentions: Unlike the development of transgenic mice which use embryonic stem cells (ES) for preselection of gene insertion [33], primary embryonic pig fibroblasts were employed for transfection with a linearized hfat-1 expression plasmid. The screening for positive cells was performed in G418-containing culture medium for 10 days; 50 G418 -resistant clones were picked and analyzed via PCR for the integration of hfat-1 cDNA. Thirty clones exhibited both the expected 458 bp band of hfat-1 and the 790 bp band of the neomycin sequence (data not shown); 5 of these clones were selected for presentation in this paper and for individually performing somatic SCNT in the following step (Fig. 2a and b).Fig. 2


N-3 polyunsaturated fatty acids attenuates triglyceride and inflammatory factors level in hfat-1 transgenic pigs.

Liu X, Pang D, Yuan T, Li Z, Li Z, Zhang M, Ren W, Ouyang H, Tang X - Lipids Health Dis (2016)

Identification of G418-resistant clones and hfat-1 transgenic pigs. Analyse of the hfat-1 gene and neomycin gene integration into the pig genome in the G418-resistant clones were performed using a PCR assay with the primer pairs of hfat-1 identi (458 bp) (a) and neo identi (790 bp) (b). a 1–5, five G418-resistant clones, on which SCNT was individually performed in the following procedure. 6, hfat-1 plasmid as a positive control. 7–8, negative controls. b 1, hfat-1 plasmid as a positive control. 2–3, negative controls. 4–8, five G418-resistant clones. The hfat-1 gene integration into the pig genome and transcription in the transgenic pigs were analyzed using PCR (c) and RT-PCR (d), respectively, with the hfat-1 identi primer pair (458 bp). c 1–5, five hfat-1 transgenic pigs. 6, hfat-1 plasmid as a positive control. 7–8, two wild-type pigs. d 1–5, five hfat-1 transgenic pigs. 6–7, two wild-type pigs
© Copyright Policy - OpenAccess
Related In: Results  -  Collection

License 1 - License 2
Show All Figures
getmorefigures.php?uid=PMC4862157&req=5

Fig2: Identification of G418-resistant clones and hfat-1 transgenic pigs. Analyse of the hfat-1 gene and neomycin gene integration into the pig genome in the G418-resistant clones were performed using a PCR assay with the primer pairs of hfat-1 identi (458 bp) (a) and neo identi (790 bp) (b). a 1–5, five G418-resistant clones, on which SCNT was individually performed in the following procedure. 6, hfat-1 plasmid as a positive control. 7–8, negative controls. b 1, hfat-1 plasmid as a positive control. 2–3, negative controls. 4–8, five G418-resistant clones. The hfat-1 gene integration into the pig genome and transcription in the transgenic pigs were analyzed using PCR (c) and RT-PCR (d), respectively, with the hfat-1 identi primer pair (458 bp). c 1–5, five hfat-1 transgenic pigs. 6, hfat-1 plasmid as a positive control. 7–8, two wild-type pigs. d 1–5, five hfat-1 transgenic pigs. 6–7, two wild-type pigs
Mentions: Unlike the development of transgenic mice which use embryonic stem cells (ES) for preselection of gene insertion [33], primary embryonic pig fibroblasts were employed for transfection with a linearized hfat-1 expression plasmid. The screening for positive cells was performed in G418-containing culture medium for 10 days; 50 G418 -resistant clones were picked and analyzed via PCR for the integration of hfat-1 cDNA. Thirty clones exhibited both the expected 458 bp band of hfat-1 and the 790 bp band of the neomycin sequence (data not shown); 5 of these clones were selected for presentation in this paper and for individually performing somatic SCNT in the following step (Fig. 2a and b).Fig. 2

Bottom Line: Although beneficial effects of n-3 PUFAs have been observed in a number of studies, the mechanisms involved in these effects have yet to be discovered.Gas chromatography results demonstrated that the total n-3 PUFAs in the ear tissues of the transgenic founders were 2-fold higher than the wild-type pigs.The basal levels of the inflammation mediators tumor necrosis factor-α (TNF-α), monocyte chemoattractant protein-1 (MCP-1) and interleukin-6 (IL-6) in transgenic pigs were inhibited markedly compared with the wild-type pigs.

View Article: PubMed Central - PubMed

Affiliation: Jilin Provincial Model Animal Engineering Research Center, College of Animal Sciences, Jilin University, Xi'an Road, 5333#, Jilin, 130062, China.

ABSTRACT

Background: The consumption of n-3 polyunsaturated fatty acids (PUFAs) is important to human health, especially in cases of cardiovascular disease. Although beneficial effects of n-3 PUFAs have been observed in a number of studies, the mechanisms involved in these effects have yet to be discovered.

Methods: We generated hfat-1 transgenic pigs with traditional somatic cell nuclear transfer (SCNT) technology. The fatty acid composition in ear tissue of pigs were detected with gas chromatography. The cholesterol, triglycerides (TAG) and inflammation mediators in circulation were investigated.

Results: The hfat-1 transgenic pigs were developed which accumulate high levels of n-3 PUFAs than wild-types pigs. Gas chromatography results demonstrated that the total n-3 PUFAs in the ear tissues of the transgenic founders were 2-fold higher than the wild-type pigs. A lipid analysis demonstrated that the levels of TAG in the transgenic pigs were decreased significantly. The basal levels of the inflammation mediators tumor necrosis factor-α (TNF-α), monocyte chemoattractant protein-1 (MCP-1) and interleukin-6 (IL-6) in transgenic pigs were inhibited markedly compared with the wild-type pigs.

Conclusions: These results suggest that n-3 PUFAs accumulation in vivo may have beneficial effects on vascular and hfat-1 transgenic pigs may be a useful tool for investigating the involved mechanisms.

No MeSH data available.


Related in: MedlinePlus