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1.
Zhonghua Gan Zang Bing Za Zhi ; 29(2): 156-162, 2021 Feb 20.
Artigo em Chinês | MEDLINE | ID: mdl-33685085

RESUMO

Objective: To investigate the correlation between patatin-like phospholipase domain-containing 3 (PNPLA3) rs738409 and transmembrane 6 superfamily member 2 (TM6SF2) rs58542926 gene polymorphisms and the incidence of primary liver cancer in the Han population of China's Northeast region. Methods: A case-control study was used to enroll 521 patients with primary liver cancer as the case group and 164 healthy people as the control group. The case group was divided into groups with and without liver cirrhosis according to etiology. The polymerase chain reaction (PCR) method was used to detect the genetic polymorphisms of PNPLA3 rs738409 and TM6SF2 rs58542926, respectively. Results: Compared with the control group, the frequency distribution of PNPLA3 rs738409 G allele in the case group was significantly different (OR = 1.583, P = 0.001). Further grouping showed that there was no statistically significant difference between the control and hepatitis C-related liver cancer group (P = 0.161), but there were significant differences in other groups (P < 0.05). Compared with the control group, the frequency of TM6SF2 rs58542926 T allele in the case group was significantly higher than that in the control group (OR = 1.759, P = 0.048). After grouping, the frequency of CT/TT genotype in the liver cancer group combined with liver cirrhosis and the T allele frequency in the alcohol-related liver cancer group had statistically significant difference (P = 0.045 and 0.032, respectively) when compared with control group. The patients were divided into CG/GG group and CC group, and CT/TT group and CC group according to whether they carried PNPLA3 rs738409 G allele, and TM6SF2 rs58542926 T allele. Results showed that there were no statistically significant differences in liver enzyme indexes, albumin (ALB), total bilirubin (TBIL), alpha-fetoprotein (AFP) and fasting blood glucose between CG/GG group and CC group, and CT/TT group and CC group. The patients with liver cirrhosis in the case group were divided into≥7 groups and < 7 groups according to the Child-Pugh score. Results showed that there were no statistically significant difference in the Child-Pugh score between PNPLA3 rs738409 CG/GG group and CC group patients and TM6SF2 rs58542926 CT/TT group and CC group patients (P > 0.05). Conclusion: PNPLA3 rs738409 and TM6SF2 rs58542926 gene polymorphisms are correlated with the occurrence of primary liver cancer in the Han population of China's Northeast region. PNPLA3 rs738409 and TM6SF2 rs58542926 gene polymorphisms have no effect on indexes' such as liver enzymes, ALB, TBIL, AFP and FBS in primary liver cancer..


Assuntos
Neoplasias Hepáticas , Hepatopatia Gordurosa não Alcoólica , Estudos de Casos e Controles , China/epidemiologia , Predisposição Genética para Doença , Genótipo , Humanos , Lipase/genética , Neoplasias Hepáticas/genética , Proteínas de Membrana/genética , Polimorfismo de Nucleotídeo Único
2.
Aliment Pharmacol Ther ; 53(7): 830-843, 2021 04.
Artigo em Inglês | MEDLINE | ID: mdl-33565643

RESUMO

BACKGROUND: Cirrhosis develops in <10% of individuals homozygous for the C282Y variant in the homeostatic iron regulator (HFE) gene. Carriage of PCSK7:rs236918 is associated with an increased risk of cirrhosis in this population. AIM: To determine if genetic variants significantly associated with the risk of alcohol- and NAFLD-related cirrhosis also modulate the cirrhosis risk in C282Y homozygotes. METHODS: Variants in PCSK7, PNPLA3, TM6SF2, MBOAT7 and HSD17B13 were genotyped in 1319 C282Y homozygotes, from six European countries, of whom 171 (13.0%) had cirrhosis. Genotypic and allelic associations with the risk for developing cirrhosis were assessed, adjusting for age and sex. Fixed effects meta-analyses of the adjusted summary data for each country were performed. Post hoc association testing was undertaken in the 131 (76.6%) cases and 299 (26.0%) controls with available liver histology. RESULTS: Significant associations were observed between PCSK7:rs236918 (OR = 1.52 [95% CI 1.06-2.19]; P = 0.022; I2  = 0%); PNPLA3:rs738409 (OR = 1.60 [95% CI 1.22-2.11]; P = 7.37 × 10-4 ; I2  = 45.5%) and TM6SF2:rs58542926 (OR = 1.94 [95% CI 1.28-2.95]; P = 1.86 × 10-3 ; I2  = 0%) and the cirrhosis risk in C282Y homozygotes. These findings remained significant in the subpopulation with available liver histology. The population-attributable fractions were 5.6% for PCSK7:rs236918, 13.8% for PNPLA3:rs738409, 6.5% for TM6SF2:rs58542926 and 24.0% for carriage of all three variants combined. CONCLUSIONS: The risk of cirrhosis associated with carriage of PCSK7:rs236918 was confirmed in this much larger population of C282Y homozygotes. In addition, PNPLA3:rs738409 and TM6SF2:rs58542926 were established as significant additional risk factors. More detailed genetic testing of C282Y homozygotes would allow risk stratification and help guide future management.


Assuntos
Hemocromatose , Hepatopatia Gordurosa não Alcoólica , Europa (Continente) , Genótipo , Humanos , Lipase/genética , Cirrose Hepática/etiologia , Cirrose Hepática/genética , Proteínas de Membrana/genética , Polimorfismo de Nucleotídeo Único , Fatores de Risco , Subtilisinas
3.
Int J Biol Macromol ; 173: 1-12, 2021 Mar 15.
Artigo em Inglês | MEDLINE | ID: mdl-33476612

RESUMO

The catalytic mechanism of most lipases involves a step called "interfacial activation" which significantly increases lipases activity beyond the critical micellar concentration (CMC) of substrate. In the present study, Rhizopus chinensis lipase (RCL) was used as a research model to explore the mechanism of lipase interfacial activation beyond the CMC. Molecular dynamic (MD) simulations indicated the open- and closed-lid transitions and revealed that Phe113 was the critical site for RCL activation by its dynamic flipping. Such putative switch affecting interfacial activation has not been reported in lipase so far. The function of Phe113 was subsequently verified by mutation experiments. The F113W mutant increases the lipase catalytic efficiency (1.9 s-1·µM-1) to 280% at the optimum temperature (40 °C) and pH 8.5 with the addition of 0.12 µg protein in the 200 µL reaction system. MD simulations indicated that the fast flipping rate from the closed to the open state, the high open state proportion, and the exposure of the catalytic triad are the main reasons for the lipase activation. The mutual corroboration of simulations and site-directed mutagenesis results revealed the vital role of Phe113 in the lipase activation.


Assuntos
Butiratos/química , Proteínas Fúngicas/química , Lipase/química , Fenilalanina/química , Rhizopus/química , Sítios de Ligação , Biocatálise , Butiratos/metabolismo , Clonagem Molecular , Escherichia coli/genética , Escherichia coli/metabolismo , Proteínas Fúngicas/genética , Proteínas Fúngicas/metabolismo , Expressão Gênica , Vetores Genéticos/química , Vetores Genéticos/metabolismo , Concentração de Íons de Hidrogênio , Cinética , Lipase/genética , Lipase/metabolismo , Simulação de Dinâmica Molecular , Fenilalanina/metabolismo , Ligação Proteica , Conformação Proteica em alfa-Hélice , Conformação Proteica em Folha beta , Domínios e Motivos de Interação entre Proteínas , Proteínas Recombinantes/química , Proteínas Recombinantes/genética , Proteínas Recombinantes/metabolismo , Rhizopus/enzimologia , Especificidade por Substrato , Temperatura , Termodinâmica , Água/química
4.
Int J Mol Sci ; 22(3)2021 Jan 27.
Artigo em Inglês | MEDLINE | ID: mdl-33513710

RESUMO

Stimulation of thermogenesis in brown adipose tissue (BAT) could have far-reaching health benefits in combatting obesity and obesity-related complications. Apolipoprotein A-IV (ApoA-IV), produced by the gut and the brain in the presence of dietary lipids, is a well-known short-term satiating protein. While our previous studies have demonstrated reduced diet-induced thermogenesis in ApoA-IV-deficient mice, it is unclear whether this reduction is due to a loss of peripheral or central effects of ApoA-IV. We hypothesized that central administration of ApoA-IV stimulates BAT thermogenesis and that sympathetic and sensory innervation is necessary for this action. To test this hypothesis, mice with unilateral denervation of interscapular BAT received central injections of recombinant ApoA-IV protein or artificial cerebrospinal fluid (CSF). The effects of central ApoA-IV on BAT temperature and thermogenesis in mice with unilateral denervation of the intrascapular BAT were monitored using transponder probe implantation, qPCR, and immunoblots. Relative to CSF, central administration of ApoA-IV significantly increased temperature and UCP expression in BAT. However, all of these effects were significantly attenuated or prevented in mice with unilateral denervation. Together, these results clearly demonstrate that ApoA-IV regulates BAT thermogenesis centrally, and this effect is mediated through sympathetic and sensory nerves.


Assuntos
Tecido Adiposo Marrom/fisiologia , Apolipoproteínas A/administração & dosagem , Regulação da Expressão Gênica/efeitos dos fármacos , Sistema Nervoso Simpático/fisiologia , Termogênese/efeitos dos fármacos , Proteínas Quinases Ativadas por AMP/genética , Proteínas Quinases Ativadas por AMP/metabolismo , Tecido Adiposo Marrom/enzimologia , Tecido Adiposo Marrom/metabolismo , Animais , Apolipoproteínas A/deficiência , Peptídeo Relacionado com Gene de Calcitonina/genética , Peptídeo Relacionado com Gene de Calcitonina/metabolismo , Regulação da Expressão Gênica/genética , Lipase/genética , Lipase/metabolismo , Masculino , Camundongos , Camundongos Endogâmicos C57BL , Proteínas Recombinantes , Terceiro Ventrículo/fisiologia , Tirosina 3-Mono-Oxigenase/metabolismo , Proteína Desacopladora 1/genética , Proteína Desacopladora 1/metabolismo
5.
Sheng Wu Gong Cheng Xue Bao ; 37(1): 88-99, 2021 Jan 25.
Artigo em Chinês | MEDLINE | ID: mdl-33501792

RESUMO

The formation of most proteins consists of two steps: the synthesis of precursor proteins and the synthesis of functional proteins. In these processes, propeptides play important roles in assisting protein folding or inhibiting its activity. As an important polypeptide chain coded by a gene sequence in lipase gene, propeptide usually functions as an intramolecular chaperone, assisting enzyme molecule folding. Meanwhile, some specific sites on propeptide such as glycosylated sites, have important effect on the activity, stability in extreme environment, methanol resistance and the substrate specificity of the lipase. Studying the mechanism of propeptide-mediated protein folding, as well as the influence of propeptide on lipases, will allow to regulate lipase by alternating the propeptide folding behavior and in turn pave new ways for protein engineering research.


Assuntos
Lipase , Dobramento de Proteína , Lipase/genética , Lipase/metabolismo , Chaperonas Moleculares/genética , Chaperonas Moleculares/metabolismo , Precursores de Proteínas , Especificidade por Substrato
6.
Artigo em Inglês | MEDLINE | ID: mdl-33075494

RESUMO

MicroRNA-221-3p (miR-221-3p) is associated with both metabolic diseases and cancers. However, its role in terminal adipocyte differentiation and lipid metabolism are uncharacterized. miR-221-3p or its inhibitor was transfected into differentiating or mature human adipocytes. Triglyceride (TG) content and adipogenic gene expression were monitored, global lipidome analysis was carried out, and mechanisms underlying the effects of miR-221-3p were investigated. Finally, cross-talk between miR-221-3p expressing adipocytes and MCF-7 breast carcinoma (BC) cells was studied, and miR-221-3p expression in tumor-proximal adipose biopsies from BC patients analyzed. miR-221-3p overexpression inhibited terminal differentiation of adipocytes, as judged from reduced TG storage and gene expression of the adipogenic markers SCD1, GLUT4, FAS, DGAT1/2, AP2, ATGL and AdipoQ, whereas the miR-221-3p inhibitor increased TG storage. Knockdown of the predicted miR-221-3p target, 14-3-3γ, had similar antiadipogenic effects as miR-221-3p overexpression, indicating it as a potential mediator of mir-221-3p function. Importantly, miR-221-3p overexpression inhibited de novo lipogenesis but increased the concentrations of ceramides and sphingomyelins, while reducing diacylglycerols, concomitant with suppression of sphingomyelin phosphodiesterase, ATP citrate lyase, and acid ceramidase. miR-221-3p expression was elevated in tumor proximal adipose tissue from patients with invasive BC. Conditioned medium of miR-221-3p overexpressing adipocytes stimulated the invasion and proliferation of BC cells, while medium of the BC cells enhanced miR-221-3p expression in adipocytes. Elevated miR-221-3p impairs adipocyte lipid storage and differentiation, and modifies their ceramide, sphingomyelin, and diacylglycerol content. These alterations are relevant for metabolic diseases but may also affect cancer progression.


Assuntos
Adipócitos/metabolismo , Adipogenia/genética , Neoplasias da Mama/genética , Regulação Neoplásica da Expressão Gênica , Gotículas Lipídicas/metabolismo , MicroRNAs/genética , Proteínas 14-3-3/genética , Proteínas 14-3-3/metabolismo , Adipócitos/patologia , Adiponectina/genética , Adiponectina/metabolismo , Tecido Adiposo/metabolismo , Tecido Adiposo/patologia , Adulto , Idoso , Neoplasias da Mama/metabolismo , Neoplasias da Mama/patologia , Estudos de Casos e Controles , Diferenciação Celular , Proliferação de Células , Ceramidas/classificação , Ceramidas/metabolismo , Diacilglicerol O-Aciltransferase/genética , Diacilglicerol O-Aciltransferase/metabolismo , Proteínas de Ligação a Ácido Graxo/genética , Proteínas de Ligação a Ácido Graxo/metabolismo , Feminino , Humanos , Lipase/genética , Lipase/metabolismo , Células MCF-7 , Glândulas Mamárias Humanas/metabolismo , Glândulas Mamárias Humanas/patologia , MicroRNAs/agonistas , MicroRNAs/antagonistas & inibidores , MicroRNAs/metabolismo , Pessoa de Meia-Idade , Transdução de Sinais , Esfingolipídeos/classificação , Esfingolipídeos/metabolismo , Estearoil-CoA Dessaturase/genética , Estearoil-CoA Dessaturase/metabolismo , Triglicerídeos/classificação , Triglicerídeos/metabolismo , Receptor fas/genética , Receptor fas/metabolismo
7.
Int J Biol Macromol ; 170: 61-70, 2021 Feb 15.
Artigo em Inglês | MEDLINE | ID: mdl-33358947

RESUMO

The increasing use of sustainable manufacturing technologies in the industry presents a constant challenge for the development of suitable biocatalysts. Traditionally, improved biocatalysts are developed either using protein engineering (PE) or enzyme immobilization (EI). However, these approaches are usually not simultaneously applied. In this work, we designed and validated an enzyme improvement platform, Immobilized Biocatalyst Engineering (IBE), which simultaneously integrates PE and EI, with a unique combination of improvement through amino acid substitutions and attachment to a support material, allowing to select variants that would not be found through single or subsequent PE and EI improvement strategies. Our results show that there is a significant difference on the best performing variants identified through IBE, when compared to those that could be identified as soluble enzymes and then immobilized, especially when evaluating variants with low enzyme as soluble enzymes and high activity when immobilized. IBE allows evaluating thousands of variants in a short time through an integrated screening, and selection can be made with more information, resulting in the detection of highly stable and active heterogeneous biocatalysts. This novel approach can translate into a higher probability of finding suitable biocatalysts for highly demanding processes.


Assuntos
Biocatálise , Enzimas Imobilizadas , Ensaios de Triagem em Larga Escala/métodos , Engenharia de Proteínas/métodos , Bacillus subtilis/enzimologia , Bacillus subtilis/genética , Proteínas de Bactérias/genética , Proteínas de Bactérias/metabolismo , Biblioteca Gênica , Lipase/genética , Lipase/metabolismo , Modelos Moleculares , Mutagênese , Estudo de Prova de Conceito , Conformação Proteica , Proteínas Recombinantes de Fusão/metabolismo , Dióxido de Silício , Solubilidade , Temperatura
8.
Sci Rep ; 10(1): 22063, 2020 12 16.
Artigo em Inglês | MEDLINE | ID: mdl-33328564

RESUMO

A novel cold-active true lipase from Pseudomonas sp. KE38 was cloned, sequencing and expressed in E. coli by degenerate PCR and genome walking technique. The open reading frame of the cloned gene encoded a polypeptide chain of 617 amino acids with a confirmed molecular weight of 64 kD. Phylogenetic analysis of the deduced amino acid sequence of the lipase indicated that it had high similarity with lipases of subfamily Ι.3 of bacterial lipases. Recombinant lipase was purified in denatured form as inclusion bodies, which were then renatured by urea followed by dialysis. Lipase activity was determined titrimetrically using olive oil as substrate. The enzyme showed optimal activity at 25 °C, pH 8.5 and was highly stable in the presence of various metal ions and organic solvents. Low optimal temperature and high activity in the presence of methanol and ethanol make this lipase a potential candidate for transesterification reactions and biodiesel production.


Assuntos
Aclimatação , Proteínas de Bactérias , Clonagem Molecular , Temperatura Baixa , Expressão Gênica , Lipase , Pseudomonas fluorescens , Proteínas de Bactérias/biossíntese , Proteínas de Bactérias/química , Proteínas de Bactérias/genética , Proteínas de Bactérias/isolamento & purificação , Lipase/biossíntese , Lipase/química , Lipase/genética , Lipase/isolamento & purificação , Pseudomonas fluorescens/enzimologia , Pseudomonas fluorescens/genética , Proteínas Recombinantes/biossíntese , Proteínas Recombinantes/química , Proteínas Recombinantes/genética , Proteínas Recombinantes/isolamento & purificação
9.
PLoS One ; 15(12): e0243919, 2020.
Artigo em Inglês | MEDLINE | ID: mdl-33326441

RESUMO

Common variants in the hepatic lipase (LIPC) gene have been shown to be associated with plasma lipid levels; however, the distribution and functional features of rare and regulatory LIPC variants contributing to the extreme lipid phenotypes are not well known. This study was aimed to catalogue LIPC variants by resequencing the entire LIPC gene in 95 non-Hispanic Whites (NHWs) and 95 African blacks (ABs) with extreme HDL-C levels followed by in silico functional analyses. A total of 412 variants, including 43 novel variants were identified; 56 were unique to NHWs and 234 were unique to ABs. Seventy-eight variants in NHWs and 89 variants in ABs were present either in high HDL-C group or low HDL-C group. Two non-synonymous variants (p.S289F, p.T405M), found in NHWs with high HDL-C group were predicted to have damaging effect on LIPC protein by SIFT, MT2 and PP2. We also found several non-coding variants that possibly reside in the circRNA and lncRNA binding sites and may have regulatory potential, as identified in rSNPbase and RegulomeDB databases. Our results shed light on the regulatory nature of rare and non-coding LIPC variants as well as suggest their important contributions in affecting the extreme HDL-C phenotypes.


Assuntos
HDL-Colesterol/sangue , LDL-Colesterol/sangue , Lipase/genética , Afro-Americanos , Alelos , Sítios de Ligação/genética , Colesterol/sangue , Colesterol/genética , Proteínas de Transferência de Ésteres de Colesterol/sangue , Proteínas de Transferência de Ésteres de Colesterol/genética , HDL-Colesterol/genética , LDL-Colesterol/genética , Grupo com Ancestrais do Continente Europeu , Feminino , Genótipo , Humanos , Íntrons/genética , Lipase/ultraestrutura , Metabolismo dos Lipídeos/genética , Lipídeos/sangue , Lipídeos/genética , Masculino , Polimorfismo de Nucleotídeo Único , Ligação Proteica/genética , Conformação Proteica , RNA Circular/sangue , RNA Circular/genética , RNA Longo não Codificante/genética , Triglicerídeos/sangue , Triglicerídeos/genética
10.
Phytochemistry ; 180: 112538, 2020 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-33091779

RESUMO

Storage lipid mobilization by lipases and lipoxygenases (LOXs) in response to developmental cues take place during seed germination. After rice grain milling, the endogenous lipases and LOXs present in the bran fraction come in contact with the storage lipid reserve or triacylglycerol (TAG). Lipases catalyze the hydrolysis of TAGs to non-esterified fatty acids (NEFAs) and glycerol. The NEFAs, especially linoleic acid (18:2) produced, are further subjected to oxidative rancidity via peroxidation reaction catalyzed by LOXs. This results in the production of conjugated hydroperoxides of 18:2 that influence the off-flavors in rice bran lipids. The aim of this study is to understand how lipid mobilization and expression of lipase and LOX genes occur in the bran of germinating rice grains (Oryza sativavar. Pusa Basmati 1). Our results show that the primary source of storage lipids in bran is TAG, and its mobilization starts at 4 days after imbibition (4 DAI). Using publically available RNA-seq data and phylogeny analyses, we selected a total of 18 lipase and 16 LOX genes in rice for their expression profiles during onset of lipid mobilization. Gene expression analyses revealed OsLip1, OsLip9, and OsLip13; and OsLOX3 and OsLOX14 as the predominantly expressed genes in bran of germinating rice grains. This study explores two important events in the germinating rice grains, namely, mobilization of storage lipids and expression pattern of lipase and LOX genes. The information generated in this study can be used to efficiently manipulate the genes to enhance the shelf-stability of bran lipid reserve.


Assuntos
Oryza , Germinação , Lipase/genética , Lipídeos , Lipoxigenase/genética , Lipoxigenases , Oryza/genética
11.
Zhonghua Yi Xue Yi Chuan Xue Za Zhi ; 37(10): 1136-1138, 2020 Oct 10.
Artigo em Chinês | MEDLINE | ID: mdl-32924119

RESUMO

OBJECTIVE: To explore the genetic basis for a child with ichthyosis. METHODS: High-throughput sequencing was carried out to detect genomic copy number variants (CNVs) and variant of the medical exome. Candidate variant was verified by Sanger sequencing. RESULTS: No disease-related CNV was identified in the patient. High-throughput sequencing revealed that the child has carried compound heterozygous variants of the PNPLA1 gene, including a previously known pathogenic c.100G>A (p.Ala34Thr) mutation and a novel c.56C>A (p.Ser19x) variant which was predicted to be a pathogenic according to the ACMG guidelines. Sanger sequencing confirmed both variants in the child. Her father and mother were found to be heterozygous carriers for the c.56C>A (p.Ser19x) and c.100G>A (p.Ala34Thr) variants, respectively. CONCLUSION: The compound heterozygous c.100G>A and c.56C>A variants of the PNPLA1 gene probably underlay the ichthyosis in this child.


Assuntos
Ictiose Lamelar , Lipase/genética , Criança , Feminino , Heterozigoto , Sequenciamento de Nucleotídeos em Larga Escala , Humanos , Ictiose Lamelar/genética , Mutação
12.
Sheng Wu Gong Cheng Xue Bao ; 36(8): 1556-1567, 2020 Aug 25.
Artigo em Chinês | MEDLINE | ID: mdl-32924354

RESUMO

Improving the thermal stability of enzymes is a hot and difficult point in the field of biocatalysis. Compared with the traditional directed evolution, computational assisted rational design is more efficient, and is widely used in enzyme engineering. Using Bacillus subtilis LipA as the model protein, the structure cavity of the enzyme was analyzed by Rosetta-VIP design, the mutation which was beneficial to the filling of the structure cavity (ΔΔE<0) was selected, followed by the solvent accessible surface area and evolutionary conservation analysis. The thermal stabilities of six out of sixteen designed single-point mutants were improved, with a maximum ΔTm value of 3.18 °C. These six mutations were further used for iterative combination mutation, the maximum ΔTm of the two-point and three-point combination mutants were 4.04 °C and 5.13 °C, respectively. The Tm of the four-point combination mutant M11 (F17A/L114P/I135V/M137L) was increased by 7.30 °C. The Tm of the six-point combination mutant M10 (F17A/V74I/L114P/I135V/M137A/I157L) was increased by 7.43 °C. The thermal stability of mutation with lower energy value, reduced accessible surface area, while conformed to evolutionary conservatism, was more likely to be improved. Therefore, the multiple virtual screening strategy based on the enzyme structure cavity filling, solvent accessible surface area and amino acid sequence conservation analysis can effectively improve the thermal stability of enzyme.


Assuntos
Bacillus subtilis , Biologia Computacional , Estabilidade Enzimática , Lipase , Bacillus subtilis/enzimologia , Bacillus subtilis/genética , Lipase/química , Lipase/genética , Lipase/metabolismo , Mutação
13.
BMC Med Genet ; 21(1): 163, 2020 08 18.
Artigo em Inglês | MEDLINE | ID: mdl-32811452

RESUMO

BACKGROUND: Nonalcoholic fatty liver disease (NAFLD) is one of the most common causes of chronic liver disease worldwide. Current studies have shown that PNPLA3 (Patatin-like phospholipase domain containing 3) rs738409 G/C gene polymorphism is associated with adult nonalcoholic fatty liver disease [1, 2].But there is no consensus on the relationship between PNPLA3 rs738409 G/C gene polymorphism and children NAFLD due to differences in population samples. To this end, a meta-analysis of published research is conducted to comprehensively assess the relationship between PNPLA3 gene polymorphism and NAFLD in children. METHODS: We searched MEDLINE, PubMed, EMBASE, and CENTRAL databases from inception to May 2019. Case-control studies assessing the relationship between PNPLA3 rs738409 G/C gene polymorphism with non-alcoholic fatty liver disease in children were selected according to inclusion and exclusion criteria. Random effects model was used to quantify the association between the PNPLA3 rs738409 G/C gene polymorphism and the susceptibility of children's NAFLD. Fixed effects model was used to quantify the relationship between the PNPLA3 rs738409 G/C gene polymorphism and the severity of NAFLD in children. The Stata 12.0 software was employed for data analysis. RESULTS: A total of nine case-control studies were included in this meta-analysis containing data of 1173 children with NAFLD and 1792 healthy controls. Five studies compared NAFLD children and non-NAFLD healthy populations. Statistical analysis showed that PNPLA3 gene polymorphism was significantly associated with children's NAFLD in the allele contrast, dominant, recessive and over dominant models (G vs C,OR = 3.343, 95% CI = 1.524-7.334; GG + GC vs CC,OR = 3.157, 95% CI = 1.446-6.892;GG vs GC + CC,OR = 5.692, 95% CI = 1.941-16.689; GG + CC vs GC,OR = 2.756, 95% CI = 1.729-4.392). Four case-control studies compared Children with nonalcoholic fatty liver (NAFL) and children with nonalcoholic steatohepatitis (NASH). The results showed that the PNPLA3 gene polymorphism was also significantly associated with the severity of NAFLD in children in recessive gene model (GG vs GC + CC,OR = 14.43, 95% CI = 5.985-34.997); The Egger's test revealed no significant publication bias. CONCLUSIONS: Meta-analysis showed that PNPLA3 gene polymorphism was significantly associated with susceptibility and severity of NAFLD in children.


Assuntos
Estudos de Associação Genética , Predisposição Genética para Doença , Lipase/genética , Proteínas de Membrana/genética , Hepatopatia Gordurosa não Alcoólica/genética , Polimorfismo de Nucleotídeo Único/genética , Criança , Feminino , Humanos , Masculino , Razão de Chances , Viés de Publicação
14.
PLoS One ; 15(8): e0237430, 2020.
Artigo em Inglês | MEDLINE | ID: mdl-32841307

RESUMO

BACKGROUND & AIMS: Given ongoing challenges in non-invasive non-alcoholic liver disease (NAFLD) diagnosis, we sought to validate an ALT-based NAFLD phenotype using measures readily available in electronic health records (EHRs) and population-based studies by leveraging the clinical and genetic data in the Million Veteran Program (MVP), a multi-ethnic mega-biobank of US Veterans. METHODS: MVP participants with alanine aminotransferases (ALT) >40 units/L for men and >30 units/L for women without other causes of liver disease were compared to controls with normal ALT. Genetic variants spanning eight NAFLD risk or ALT-associated loci (LYPLAL1, GCKR, HSD17B13, TRIB1, PPP1R3B, ERLIN1, TM6SF2, PNPLA3) were tested for NAFLD associations with sensitivity analyses adjusting for metabolic risk factors and alcohol consumption. A manual EHR review assessed performance characteristics of the NAFLD phenotype with imaging and biopsy data as gold standards. Genetic associations with advanced fibrosis were explored using FIB4, NAFLD Fibrosis Score and platelet counts. RESULTS: Among 322,259 MVP participants, 19% met non-invasive criteria for NAFLD. Trans-ethnic meta-analysis replicated associations with previously reported genetic variants in all but LYPLAL1 and GCKR loci (P<6x10-3), without attenuation when adjusted for metabolic risk factors and alcohol consumption. At the previously reported LYPLAL1 locus, the established genetic variant did not appear to be associated with NAFLD, however the regional association plot showed a significant association with NAFLD 279kb downstream. In the EHR validation, the ALT-based NAFLD phenotype yielded a positive predictive value 0.89 and 0.84 for liver biopsy and abdominal imaging, respectively (inter-rater reliability (Cohen's kappa = 0.98)). HSD17B13 and PNPLA3 loci were associated with advanced fibrosis. CONCLUSIONS: We validate a simple, non-invasive ALT-based NAFLD phenotype using EHR data by leveraging previously established NAFLD risk-associated genetic polymorphisms.


Assuntos
Alanina Transaminase/metabolismo , Hepatopatia Gordurosa não Alcoólica/patologia , 17-Hidroxiesteroide Desidrogenases/genética , Abdome/diagnóstico por imagem , Proteínas Adaptadoras de Transdução de Sinal/genética , Idoso , Alanina Transaminase/genética , Registros Eletrônicos de Saúde , Feminino , Loci Gênicos , Predisposição Genética para Doença , Variação Genética , Humanos , Lipase/genética , Fígado/patologia , Lisofosfolipase/genética , Masculino , Proteínas de Membrana/genética , Pessoa de Meia-Idade , Hepatopatia Gordurosa não Alcoólica/etnologia , Hepatopatia Gordurosa não Alcoólica/genética , Fenótipo , Fatores de Risco , Veteranos
15.
Nutr Metab Cardiovasc Dis ; 30(9): 1564-1572, 2020 08 28.
Artigo em Inglês | MEDLINE | ID: mdl-32636123

RESUMO

BACKGROUND AND AIMS: We previously demonstrated that children with Down syndrome (DS) exhibited a greater risk of steatosis than the general pediatric population. This trend was independent of obese phenotype, thus suggesting a role of genetic predisposition. Therefore, we investigated the prevalence of non-alcoholic fatty liver disease (NAFLD) and metabolic syndrome (MetS) in function of genetic susceptibility and adipocytokine levels in children with DS. METHODS AND RESULTS: A total of 84 Caucasian children with DS (age range 5-17 years), were included in this study. For all children, we collected data on anthropometric and biochemical parameters, and liver ultrasound (US). We also measured adipocytokines circulating levels and specific polymorphisms closed to NAFLD. We found a prevalence of 64.3% of liver steatosis at US, with a severe steatosis of about 4% in children with DS. The presence of steatosis in children with DS was associated with the presence of patatin-like phospholipase domain-containing 3 (PNPLA3) rs738409 variant, which also correlated with interleukin (IL)-6 levels. Moreover, we found that the 52.4% had a waist circumference > 90th percentile, 21.4% were hypertensive, 7.14% had hyperglycemia, 9.5% had hypertriglyceridemia, and 17.9% showed high-density lipoprotein cholesterol ≤ 40 mg/dl. Finally, the IL-6 and adiponectin levels correlated with steatosis, and several adipocytokines correlated with single MetS traits in children with DS. CONCLUSION: The present study explores for the first time potential pathomechanisms connecting pediatric NAFLD and MetS in DS. We found that the PNPLA3 variant is associated with steatosis, but not with MetS, in children with DS.


Assuntos
Síndrome de Down/genética , Lipase/genética , Proteínas de Membrana/genética , Síndrome Metabólica/genética , Hepatopatia Gordurosa não Alcoólica/genética , Polimorfismo de Nucleotídeo Único , Adiponectina/sangue , Adolescente , Fatores Etários , Biomarcadores/sangue , Glicemia/metabolismo , Criança , Pré-Escolar , Síndrome de Down/sangue , Síndrome de Down/diagnóstico , Síndrome de Down/epidemiologia , Feminino , Predisposição Genética para Doença , Humanos , Interleucina-6/sangue , Lipídeos/sangue , Masculino , Síndrome Metabólica/sangue , Síndrome Metabólica/diagnóstico , Síndrome Metabólica/epidemiologia , Hepatopatia Gordurosa não Alcoólica/sangue , Hepatopatia Gordurosa não Alcoólica/diagnóstico , Hepatopatia Gordurosa não Alcoólica/epidemiologia , Fenótipo , Prevalência , Medição de Risco , Fatores de Risco , Roma/epidemiologia
16.
PLoS One ; 15(6): e0234465, 2020.
Artigo em Inglês | MEDLINE | ID: mdl-32544194

RESUMO

Obesity leads a crucial importance in metabolic disorders, as well as type 2 diabetes mellitus. Our present study was designed to assess the potential role of irisin, adiponectin, leptin and gene polymorphism of PNPLA3, leptin and adiponectin as predictive markers of diabetes associated with obesity. One hundred eighty subjects were distributed to three groups including; healthy non-diabetic non obese volunteers as a control group, diabetic non obese group, and diabetic obese group (n = 60 for each group). Fasting blood samples of all groups were collected to determine fasting blood glucose, insulin levels, insulin resistance, total cholesterol, high density lipoprotein cholesterol (HDL-C), low density lipoprotein cholesterol (LDL-C), triacylglycerol, irisin, adiponectin, leptin; as well as, polymorphism of PNPLA3, adiponectin and leptin. The results showed that glucose, insulin resistance, total cholesterol, irisin, leptin, LDL-C, triacylglycerol concentrations were significantly increased, however, insulin, HDL-C, adiponectin were significantly decreased in diabetic obese patients in relation to diabetic non-obese patients as well as in healthy volunteers. The polymorphism of PNPLA3 rs738409 was linearly related to irisin and leptin but was not related with circulating concentrations of adiponectin. We concluded that increased irisin and leptin levels can predict the insulin resistance in obese patients. Moreover, patients who have mutant genotype of PNPLA3 I148 gene (rs738409) C>G, ADIPOQ gene (rs266729) G>C and LEP gene (rs2167270) G>A showed a significant higher susceptibility rate for DM in obese people than those with wild type. This could be considered as an adjustable retort to counter the impact of obesity on glucose homeostasis.


Assuntos
Adiponectina/genética , Diabetes Mellitus Tipo 2/etiologia , Predisposição Genética para Doença , Resistência à Insulina/genética , Leptina/genética , Lipase/genética , Proteínas de Membrana/genética , Obesidade/complicações , Obesidade/genética , Adiponectina/sangue , Adulto , Feminino , Fibronectinas/sangue , Fibronectinas/genética , Marcadores Genéticos , Humanos , Leptina/sangue , Lipase/sangue , Masculino , Proteínas de Membrana/sangue , Polimorfismo Genético , Adulto Jovem
20.
Arch Biochem Biophys ; 689: 108475, 2020 08 15.
Artigo em Inglês | MEDLINE | ID: mdl-32585312

RESUMO

The unconventional yeast Yarrowia lipolytica is known as a producer of extracellular lipases. Here we overexpressed extracellular lipase (YlLip2) in yeast strain Y. lipolytica AJD ΔXΔA-Lip2 harboring the overexpression cassette of the YALI0A20350 gene under the strong hybrid promoter UAS1B16-TEF. To maintain a high level of YlLip2 production, two extracellular proteases of Y. lipolytica, AEPp and AXPp, were deleted. The purified recombinant YlLip2 presented optimal catalytic activities at 37 °C and pH 8.0. The effect of two lipopeptide biosurfactants, i.e., amphisin produced by Pseudomonas fluorescens DSS73 and viscosinamide secreted by P. fluorescens DR54, on the conformation and activity of YlLip2 was evaluated using spectral methods, surface tension, and the enzyme activity assay. YlLip2 demonstrated high tolerance of the tested biosurfactants and had greater activity retention after incubation with both biosurfactants. Finally, we observed that intrinsic fluorescence intensity of YlLip2 decreased significantly with increasing lipopeptides concentration ranging from 2.5 to 60 µM. Our results showed that both biosurfactants improve enzymatic activity of YlLip2 and might suggest better interaction of the substrate with the active site. These favorable characteristics make YlLip2 a prospective additive in the pharmaceutical, food, cosmetic, and detergent industries.


Assuntos
Lipase/metabolismo , Lipopeptídeos/metabolismo , Yarrowia/enzimologia , Proteínas Fúngicas/genética , Proteínas Fúngicas/metabolismo , Lipase/genética , Pseudomonas fluorescens/metabolismo , Proteínas Recombinantes/genética , Proteínas Recombinantes/metabolismo , Especificidade por Substrato , Regulação para Cima , Yarrowia/genética , Yarrowia/metabolismo
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