Objective: The aim of this study was to detect single nucleotide polymorphisms (SNP) of G protein-coupled receptor 54 (GPR54) gene and explore association of this candidate gene with reproductive traits in Jiaxing Black sows. Methods: Six pairs of primers of the gene were designed to amplify all exons thus sequences of which were detected by means of direct sequencing and then SNP loci were scanned. The effects of SNPs on total number of piglets born (TNB), number of piglets born alive (NBA), number of still born piglets (NSB), and litter weight at birth (LWB) of Jiaxing Black sows were analyzed. Results: Three SNP loci, including T3739C, C3878T and T6789C, were identified via comparison of sequencing and two genotypes (AB, BB) at each SNP site were observed. T3739C resulted in the change of amino acid ($Leu{\rightarrow}Pro$) in corresponding protein, and C3878T resulted in synonymous mutation ($Ile{\rightarrow}Ile$). Statistical results demonstrated that allele B was the preponderant allele at the three SNP loci and Genotype BB was the preponderant genotype. Meanwhile, Chi-Square test of these three SNPs indicated that all mutation sites fitted in Hardy-Weinberg equilibrium (p>0.05). For GPR54-T3739C locus, Jiaxing Black sows with genotype BB had 1.23 TNB and 1.28 NBA (p<0.01) that were more than those with genotype AB, respectively. Jiaxing Black sows that had the first two parities with genotype BB had additional 2.23 TNB, 2.27 NBA (p<0.01), and 1.94 LWB (p<0.05) compared to those with genotype AB, respectively. However, for other two loci, no significant difference was found between TNB, NBA, NSB, and LWB, and different genotypes of Jiaxing Black sows. Conclusion: In conclusion, the polymorphisms of GPR54-T3739C locus were significantly associated to TNB, NBA, and LWB and could be used as a potential genetic marker to improve reproductive function of Jiaxing black sows.
Objective: Among stress responses, the unfolded protein response (UPR) is a well-known mechanism related to endoplasmic reticulum (ER) stress. ER stress is induced by a variety of external and environmental factors such as starvation, ischemia, hypoxia, oxidative stress, and heat stress. Inositol requiring enzyme $1{\alpha}$ ($IRE1{\alpha}$)-X-box protein 1 (XBP1) is the most conserved pathway involved in the UPR and is the main component that mediates $IRE1{\alpha}$ signalling to downstream ER-associated degradation (ERAD)- or UPR-related genes. XBP1 is a transcription factor synthesised via a novel mechanism called 'frame switch splicing', and this process has not yet been studied in the horse XBP1 gene. Therefore, the aim of this study was to confirm the frame switch splicing of horse XBP1 and characterise its dynamics using Thoroughbred muscle cells exposed to heat stress. Methods: Primary horse muscle cells were used to investigate heat stress-induced frame switch splicing of horse XBP1. Frame switch splicing was confirmed by sequencing analysis. XBP1 amino acid sequences and promoter sequences of various species were aligned to confirm the sequence homology and to find conserved cis-acting elements, respectively. The expression of the potential XBP1 downstream genes were analysed by quantitative real-time polymerase chain reaction. Results: We confirmed that splicing of horse XBP1 mRNA was affected by the duration of thermal stress. Twenty-six nucleotides in the mRNA of XBP1 were deleted after heat stress. The protein sequence and the cis-regulatory elements on the promoter of horse XBP1 are highly conserved among the mammals. Induction of putative downstream genes of horse XBP1 was dependent on the duration of heat stress. We confirmed that both the mechanisms of XBP1 frame switch splicing and various binding elements found in downstream gene promoters are highly evolutionarily conserved. Conclusion: The frame switch splicing of horse XBP1 and its dynamics were highly conserved among species. These results facilitate studies of ER-stress in horse.
Yu, Ju Hyeong;Geum, Na Gyeong;Ye, Joo Ho;Jeong, Jin Boo
Korean Journal of Plant Resources
/
v.34
no.5
/
pp.411-419
/
2021
In this study, we investigated in vitro immune-enhancing and anti-obesity activity of Abelmoschus manihot roots (AMR) in mouse macrophage RAW264.7 cells and mouse adipocytes 3T3-L1 cells. AMR increased the production of immunostimulatory factors such as nitric oxide (NO), inducible nitric oxide synthase (iNOS), interleukin-1β (IL-1β), interleukin-6 (IL-6) and tumor necrosis factor-α (TNF-α) in RAW264.7 cells. The inhibition of toll like receptor (TLR) 2 and 4 blocked AMR-mediated production of immunostimulatory factors in RAW264.7 cells. In addition, the inhibition of mitogen-activated protein kinases (MAPKs) signaling pathway reduced AMR-mediated production of immunostimulatory factors. From these results, AMR is considered to have immune-enhancing activity through TLR2/4-mediated activation of MAPKs signaling pathway. In addition, AMR inhibited lipid accumulation and reduced the protein level such as CCAAT enhancer-binding protein alpha (CEBPα), peroxisome proliferator-activated receptor gamma (PPARγ), perilipin-1, adiponectin and fatty acid binding protein 4 (FABP4) associated with lipid accumulation in 3T3-L1 cells, indicating that AMR may have anti-obesity activity. Based on these results, AMR is expected to be used as a potential functional agent for immune enhancement and anti-obesity.
Go, Eun Ji;Ryu, Byung Ryeol;Yang, Su Jin;Baek, Jong Suep;Ryu, Su Ji;Kim, Hyun Bok;Lim, Jung Dae
Korean Journal of Medicinal Crop Science
/
v.28
no.6
/
pp.395-411
/
2020
Background: This study investigated the anti-obesity effect of the flavonoid rich fraction (FRF) and its constituent, rutin obtained from the leaf of Morus alba L., on the lipid accumulation mechanism in 3T3-L1 adipocyte and C57BL/6 mouse models. Methods and Results: In Oil Red O staining, FRF (1,000 ㎍/㎖) treatments showed inhibition rate of 35.39% in lipid accumulation compared to that in the control. AdipoRedTM assay indicated that the triglyceride content in 3T3-L1 adipocytes treated with FRF (1,000 ㎍/㎖) was reduced to 23.22%, and free glycerol content was increased to 106.04% that of the control. FRF and its major constituent, rutin affected mRNA gene expression. Rutin contributed to the inhibition of Sterol regulatory element binding protein-1c (SREBP-1c) gene expression, and inhibited the transcription factors SREBP-1c, peroxisome proliferator-activated receptor gamma (PPAR-γ), CCAAT/enhancer binding protein α (C/EBPα), fatty acid synthase (FAS) and acetyl-CoA carboxylase (ACC). In addition, the effect of FRF administration on obesity development in C57BL/6 mice fed high-fat diet (HFD) was investigated. FRF suppressed weight gain, and reduced liver triglyceride and leptin secretion. FRF exerted potential anti-inflammatory effects by improving insulin resistance and adiponectin levels, and could thus be used to help counteract obesity. The mRNA expressions of PPAR-γ, FAS, ACC, and CPT-1 were determined in liver tissue. Quantitative real-time PCR analysis was also performed to evaluate the expression of IL-1β, IL-6, and TNF-α in epididymal adipose tissue. Compared to the control group, mice fed the HFD showed the up-regulation in PPAR-γ, FAS, IL-6, and TNF-α genes, and down-regulation in CPT1 gene expression. FRF treatement markedly reduced the expression of PPAR-γ, FAS, IL-6, and TNF-α compared to those in HFD control, whereas increased the expression level of CPT1. Conclusions: These results suggest that the FRF and its major active constituent, rutin, can be used as effective anti-obesity agents.
Objective: The aim of the study was to evaluate the influence of polymorphic loci and other factors on milk performance and the technological properties of milk. Methods: The analysis was performed on Simmental and Holstein cows in field conditions (n = 748). Milk yield in kg, fat and protein percentage and yield were evaluated. Technological properties were evaluated by milk fermentation ability, renneting, and an alcohol test. Polymorphisms in the acyl-CoA diacylgycerol transferase 1 (DGAT1), leptin (LEP), fatty acid synthase (FASN), stearoyl CoA desaturase 1 (SCD1), casein beta (CSN2), casein kappa (CSN3), and lactoglobulin beta genes were genotyped, and association analysis was performed. Results: The DGAT1 AA genotype was associated with higher milk, protein and fat yields (p<0.05). The MM genotype in the LEP gene was associated with a lower protein percentage and the W allele with a higher protein percentage (p<0.05). In cows with the FASN GG genotype, the protein percentage was higher, but the A allele was associated with higher milk, protein and fat yields than the G allele. The TT genotype in SCD1 was associated with the lowest milk, protein and fat yields and with the highest milk protein percentage (p<0.01). The T allele had higher values than the C allele (p<0.05) except for fat percentage. The genotype CSN3 AA was associated with a significantly heightened milk yield; BB was associated with a high protein percentage. The effect of the alleles on the technological properties was not significant. The CSN2 BB genotype was associated with the best alcohol test (p<0.01), and the renneting order was inverse. Milk from cows with the CSN2 A1A1 genotype was best in the milk fermentation ability. CSN3 significantly affected the technological properties. Conclusion: The findings revealed the potential of some polymorphic loci for use in dairy cattle breeding and for the management of milk quality. In field research, the pivotal role of farms in milk yield, composition and technological properties was confirmed.
Objective: Vanin1 (VNN1) is a pantetheinase that can catalyze the hydrolysis of pantetheine to produce pantothenic acid and cysteamine. Our previous studies showed that VNN1 is specifically expressed in chicken liver. In this study, we aimed to investigate the roles of peroxisome proliferators activated receptor α (PPARα) and miRNA-181a-5p in regulating VNN1 gene expression in chicken liver. Methods: 5'-RACE was performed to identify the transcription start site of chicken VNN1. JASPAR and TFSEARCH were used to analyze the potential transcription factor binding sites in the promoter region of chicken VNN1 and miRanda was used to search miRNA binding sites in 3' untranslated region (3'UTR) of chicken VNN1. We used a knock-down strategy to manipulate PPARα (or miRNA-181a-5p) expression levels in vitro to further investigate its effect on VNN1 gene transcription. Luciferase reporter assays were used to explore the specific regions of VNN1 targeted by PPARα and miRNA-181a-5p. Results: Sequence analysis of the VNN1 promoter region revealed several transcription factor-binding sites, including hepatocyte nuclear factor 1α (HNF1α), PPARα, and CCAAT/enhancer binding protein α. GW7647 (a specific agonist of PPARα) increased the expression level of VNN1 mRNA in chicken primary hepatocytes, whereas knockdown of PPARα with siRNA increased VNN1 mRNA expression. Moreover, the predicted PPARα-binding site was confirmed to be necessary for PPARα regulation of VNN1 gene expression. In addition, the VNN1 3'UTR contains a sequence that is completely complementary to nucleotides 1 to 7 of miRNA-181a-5p. Overexpression of miR-181a-5p significantly decreased the expression level of VNN1 mRNA. Conclusion: This study demonstrates that PPARα is an important transcriptional activator of VNN1 gene expression and that miRNA-181a-5p acts as a negative regulator of VNN1 expression in chicken hepatocytes.
The aim of the present study was to investigate the mechanical properties of melt-injected poly lactic acid (PLA) composites with β-tricalcium phosphate (β-TCP). The PLA mixed with calcined PLA/β-TCP powder to be contents of 0, 10, 30, 50 wt%, respectively, was dissolved in chloroform solvent under stirring for 24 h. Then the liquid mixtures were dropped into ethanol to extract solvent. After drying, the well-dispersed PLA/β-TCP composites were granulated and melt-injected to prepare specimens for various mechanical testing. PLA/β-TCP induced the precipitation of an apatite bone-mineral phase on the surface after immersion in a human simulated body fluid (SBF) for 90 days, showing in bioactivity. Mean various mechanical properties PLA/β-TCP composite were increased up to 10-30 wt% with significantly in part and composite were decreased 50 wt% of showing in mechanical properties. In conclusion, Over 30 wt% addition of β-TCP to PLA may be not advisable to improve the mechanical properties of melt-injected polymeric composites. Results indicated that β-TCP can be used considered as potential reinforcing agent for increasing mechanical properties for PLA. Therefore, it was suggest that the additional effects of β-TCP and research on a wide range of substances.
Lee, Jihye;Jung, Eun Mi;Lee, Eunhong;Jang, Gwi Yeong;Seo, Kyung Hye;Kim, Mi Ryeo;Jung, Ji Wook
The Korea Journal of Herbology
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v.36
no.2
/
pp.1-9
/
2021
Objectives : Gossypium arboreum (cotton) is traditionally used to treat various health disorders. However, anti-amnesic effect of G. arboreum has not been reported. The objective of this study was to investigate in-vivo the anti-amnesic effects along with in vitro antioxidant and acetylcholinesterase (AChE) inhibition potential in G. arboreum seed essential oil. Methods : The essential oil of G. arboreum obtained by solid phase microextraction (SPME) techniques were identified by gas chromatography-mass spectroscopy (GC-MS). 2,2-diphenyl-1-picrylhydrazyl (DPPH) and 2,2'-azino-bis-(3-ethylbenzothiazoline-6-sulfonic acid) (ABTS) assay were performed to determine the antioxidant activity at various concentrations (312.5, 625, 1250, 2500, 5000, 10000 ㎍/㎖. Y-maze, passive avoidance and Morris water maze tests were carried out to evaluate improved effect on scopolamine (1 mg/kg)-induced memory dysfunction at the dose level of 50, 100 and 200 mg/kg. Donepezil (5 mg/kg) was used as a positive drug control. We performed acetylcholinesterase (AChE) activity assay in ex vivo. Results : Five volatile compounds were identified in G. arboreum. The assays of DPPH and ABTS revealed that G. arboreum increased antioxidant activity in a dose-dependent manner. G. arboreum ameliorated the percent of spontaneous alternation in the Y-maze test, shortened step-through latency in the passive avoidance test, and increased swimming time in the target zone in the Morris water maze test. In addition, G. arboreum inhibited the AChE activity. Conclusions : Based on these findings, G. arboreum may aid in the prevention and treatment of learning and memory-deficit disorders through antioxidant and AChE inhibitory activities.
Lim, Seul Ki;Lee, Jieun;Park, Sung Soo;Kim, Sun Yong;Park, Sang Min;Mok, Ji Ye;Chang, Hyunah;Choi, Hak-Jong
Microbiology and Biotechnology Letters
/
v.50
no.1
/
pp.135-146
/
2022
Obesity is closely associated with profound dyslipidemia, insulin resistance, and fatty liver disease. Recent reports have suggested that alterations in gut microbiota can be linked to diet-induced obesity. In this study, the anti-obesity effects of Weissella confusa WIKIM51 isolated from kimchi were investigated, as evidenced by: i) reduced lipid accumulation and downregulated adipogenesis-related genes in 3T3-L1 adipocytes; ii) suppressed gains in body weight and epididymal fat mass; iii) reduced serum lipid levels, for example, triglyceride and total cholesterol; iv) increased serum adiponectin levels and reduced serum leptin levels; v) downregulated lipogenesis and upregulated β-oxidation-related genes in the epididymal fat; and vi) altered microbial communities. The collective evidence indicate the potential value of W. confusa WIKIM51 as a functional food supplement for the prevention and amelioration of obesity.
This study investigated the antioxidant and anti-inflammatory effects of seed ethanol extracts from Rubus coreanus Miquel (SERC). To investigate the antioxidant activity, total polyphenol and flavonoid content, ABTS and DPPH radical scavenging activity, and reducing power were measured. The total polyphenol and flavonoid contents in seed ethanol extracts of R. coreanus Miq. were 4.09 ㎍ gallic acid equivalents (GAE)/mg and 16.25 ㎍ quercetin equivalents (QE)/mg, respectively. DPPH and ABTS radical scavenging activity showed concentration-dependent scavenging activity, and the RC50 values of SERC were 26.68 ㎍/mL and 39.30 ㎍/mL, respectively. Moreover, the ferric reducing antioxidant power (FRAP) assay was performed to assess the reducing power, and SERC showed 0.61 ± 0.01 mM FeSO4 E/mg. To measure the anti-inflammatory effect, the cytotoxicity and nitric oxide (NO) production inhibitory efficacy in RAW 264.7 cells was confirmed. SERC showed a NO production inhibitory effect at 500 ㎍/mL without cytotoxicity. As a result of verifying the antioxidant and anti-inflammatory activity using SERC, its potential as an antioxidant and anti-inflammatory material was confirmed.
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