• Title/Summary/Keyword: circulating miRNA

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Troglitazone Regulates white Adipose Tissue Metabolism by Activating Genes Involved in Fatty Acid ${\beta}$-Oxidation in High Fat Diet-fed C57BL/6J Mice

  • Jeong, Sun-Hyo;Yoon, Mi-Chung
    • Biomedical Science Letters
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    • v.12 no.4
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    • pp.319-327
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    • 2006
  • This study aimed to determine whether troglitazone stimulates genes related to fatty acid ${\beta}$-oxidation, leading to modulation of white adipose tissue (WAT) metabolism in high fat diet-fed mice. Female C57BL/6J mice were randomly divided into two groups (n=10/group). After they received either a high fat diet or the same high fat diet supplemented with troglitazone for 4 weeks, the effects of troglitazone on gene expression and physiology of WAT were measured using Northern, histological and serological analyses. Administration of troglitazone induced the expression of genes involved in mitochondrial and peroxisomal fatty acid ${\beta}$-oxidation in mesenteric WAT. Troglitazone also significantly increased uncoupling protein 2 mRNA levels. The changes in WAT gene expression were accompanied by reductions in circulating levels of free fatty acids and triglycerides as well as glucose and insulin. Histological studies showed that troglitazone treatment decreased the average size of adipocytes in mesenteric WAT. These results suggest that troglitazone-stimulated WAT expression of genes associated with fatty acid ${\beta}$-oxidation regulates WAT metabolism of high fat diet-fed mice, contributing to improvement of insulin sensitivity.

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Relationship between porcine miR-20a and its putative target low-density lipoprotein receptor based on dual luciferase reporter gene assays

  • Ding, Yueyun;Zhu, Shujiao;Wu, Chaodong;Qian, Li;Li, DengTao;Wang, Li;Wan, Yuanlang;Zhang, Wei;Yang, Min;Ding, Jian;Wu, Xudong;Zhang, Xiaodong;Gao, Yafei;Yin, Zongjun
    • Asian-Australasian Journal of Animal Sciences
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    • v.32 no.7
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    • pp.922-929
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    • 2019
  • Objective: Mutations in low-density lipoprotein receptor (LDLR), which encodes a critical protein for cholesterol homeostasis and lipid metabolism in mammals, are involved in cardiometabolic diseases, such as familial hypercholesterolemia in pigs. Whereas microRNAs (miRNAs) can control LDLR regulation, their involvement in circulating cholesterol and lipid levels with respect to cardiometabolic diseases in pigs is unclear. We aimed to identify and analyze LDLR as a potential target gene of SSC-miR-20a. Methods: Bioinformatic analysis predicted that porcine LDLR is a target of SSC-miR-20a. Wild-type and mutant LDLR 3'-untranslated region (UTR) fragments were generated by polymerase chain reaction (PCR) and cloned into the pGL3-Control vector to construct pGL3 Control LDLR wild-3'-UTR and pGL3 Control LDLR mutant-3'-UTR recombinant plasmids, respectively. An miR-20a expression plasmid was constructed by inserting the porcine premiR-20a-coding sequence between the HindIII and BamHI sites in pMR-mCherry, and constructs were confirmed by sequencing. HEK293T cells were co-transfected with the miR-20a expression or pMR-mCherry control plasmids and constructs harboring the corresponding 3'-UTR, and relative luciferase activity was determined. The relative expression levels of miR-20a and LDLR mRNA and their correlation in terms of expression levels in porcine liver tissue were analyzed using reverse-transcription quantitative PCR. Results: Gel electrophoresis and sequencing showed that target gene fragments were successfully cloned, and the three recombinant vectors were successfully constructed. Compared to pMR-mCherry, the miR-20a expression vector significantly inhibited wild-type LDLR3'-UTR-driven (p<0.01), but not mutant LDLR-3'-UTR-driven (p>0.05), luciferase reporter activity. Further, miR-20a and LDLR were expressed at relatively high levels in porcine liver tissues. Pearson correlation analysis revealed that porcine liver miR-20a and LDLR levels were significantly negatively correlated (r = -0.656, p<0.05). Conclusion: LDLR is a potential target of miR-20a, which might directly bind the LDLR 3'-UTR to post-transcriptionally inhibit expression. These results have implications in understanding the pathogenesis and progression of porcine cardiovascular diseases.

Rotavirus P and G Genotypes Circulating in Kyungsangnamdo, Korea, during 2000~2001 (2000~2001년 경상남도에서 유행한 로타바이러스 유전자형)

  • So, Kyeung Jin;Lee, Mi Hyun;Ma, Sang Hyeok;Kim, Byung Chyeol;Yang, Jai Myung
    • Pediatric Infection and Vaccine
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    • v.11 no.1
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    • pp.59-72
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    • 2004
  • Purpose : Rotaviruses are the major cause of gastroenteritis in infants and young children worldwide. It is important to get the epidemiologic data of rotavirus genotype for the application of rotavirus vaccine. So we tried to investigate the distribution of rotavirus genotypes with RT-PCR. Methods : A total of 120 rotavirus latex agglutinin test positive stool samples were collected continually from 120 children from Sep. 2000 to Apr. 2001. Rotavirus P(VP4), G (VP7) genotypes were determined by RT-PCR. Results : The genotype was identified in 116 stool samples of total 120 samples(96%). The incidence of G genotype was as follow; G1 17(14.2%), G2 74(61.7%), G4 1(0.8%), G9 1(0.8%). There were four cases of multiple genotypes; G1/G2, G1/G4, G1/G9, G8/G9 and genotype of G3, G8 were not found. Twenty three(19.2%) samples were nontypeable. The incidence of P was as follow; P[4] 77(64.2%), P[6] 22(18.3%), P4/P6 12(10%), P[4]/P[8] 1(0.8%) p[8] 1(0.8%). Seven(5.9%) samples were nontypeable. Conclusion : Various combinations of G and P genotypes were observed. Most rotavirus strains were P[4]G2 62(51.74%), followed by P[6]G2 7(5.8%), and P[6]G1 7(5.8%), P[4/P[6] G1 4(3%), P[4]/P[6]G2 4(3%), P[4]G1 3(2.5%), P[8]G2 1(0.8%), P[4]G4 1(0.8%) in Kyoungsangnamdo, Korea during 2000~2001.

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