Journal: Frontiers in Pharmacology

Article Title: Effect of gentiopicroside on endogenous formaldehyde homocysteine–pathway related proteins in rats with non-alcoholic steatohepatitis

doi: 10.3389/fphar.2025.1700101

Figure Lengend Snippet: Expression levels of CBS, ALDH2, AHCY, MAT1A and MTHFR proteins in liver tissues of rats in each group. (A) Expression levels of MAT1A protein in liver tissues of rats in each group (n = 3); (B) Expression levels of AHCY protein in liver tissues of rats in each group (n = 3); (C) Expression levels of MYHFR protein in liver tissues of rats in each group (n = 3); (D) Expression levels of ALDH2 protein in liver tissues of rats in each group (n = 3); (E) Expression levels of CBS protein in liver tissues of rats in each group (n = 3). Note: Data are expressed as the mean ± SD. “#” indicates that P < 0.05 compared with the normal group; “*” indicates that P < 0.05 compared with the model group.

Article Snippet: ROS, GSH, MDA, SOD, SAH, SAM, GST, CAT, GSH, HCY were obtained from Jiangsu Enzyme Immunoassay Biotechnology Co., LTD. Their item numbers are MM-88564O1, MM-20251R1, MM-2037H1, MM-20387R1, MM-50456H1, MM-0248H1, MM-21254R1, MM-20447R1, MM-20251R1 and MM-50456H1. β-actin antibody (Proteintech, 20536-1-AP); ALDH2 antibody (Proteintech, 15310-1-AP); MAT1A antibody (Proteintech, 12395-1-AP); MTHFR antibody (Proteintech, 26591-1-AP); CBS antibody (Proteintech, 14787-1-AP); AHCY antibody (Proteintech, 10757-2-AP); PVDF membrane with a thickness of 0.45 μm (IPVH00010, Millipore); Universal total RNA extraction reagent (U7431, Suzhou Youyi Landi Biotechnology Co., LTD.) Taq SYBR® Green qPCR Premix (Universal) (EG0117M, Jiangsu Bishi Mei Biotechnology Co., LTD.) All-in-One First-Strand Synthesis MasterMix (with dsDNase) was obtained from Jiangsu Bestmate Biotechnology Co., LTD. (item number (EG15133S).

Techniques: Expressing

Journal: Frontiers in Pharmacology

Article Title: Effect of gentiopicroside on endogenous formaldehyde homocysteine–pathway related proteins in rats with non-alcoholic steatohepatitis

doi: 10.3389/fphar.2025.1700101

Figure Lengend Snippet: Expression levels of CBS, ALDH2, AHCY, MAT1A and MTHFRmRNA in liver tissues of rats in each group (A) Expression levels of ALDH2 mRNA in liver tissues of rats in each group (n = 3); (B) Expression levels of MTHFRmRNA in liver tissues of rats in each group (n = 3); (C) Expression levels of CBS mRNA in liver tissues of rats in each group (n = 3); (D) Expression levels of AHCY mRNA in liver tissues of rats in each group (n = 3); (E) Expression levels of MAT1A mRNA in liver tissues of rats in each group (n = 3). Note: “#” indicates that P < 0.05 compared with the normal group; “*” indicates that P < 0.05 compared with the model group.

Article Snippet: ROS, GSH, MDA, SOD, SAH, SAM, GST, CAT, GSH, HCY were obtained from Jiangsu Enzyme Immunoassay Biotechnology Co., LTD. Their item numbers are MM-88564O1, MM-20251R1, MM-2037H1, MM-20387R1, MM-50456H1, MM-0248H1, MM-21254R1, MM-20447R1, MM-20251R1 and MM-50456H1. β-actin antibody (Proteintech, 20536-1-AP); ALDH2 antibody (Proteintech, 15310-1-AP); MAT1A antibody (Proteintech, 12395-1-AP); MTHFR antibody (Proteintech, 26591-1-AP); CBS antibody (Proteintech, 14787-1-AP); AHCY antibody (Proteintech, 10757-2-AP); PVDF membrane with a thickness of 0.45 μm (IPVH00010, Millipore); Universal total RNA extraction reagent (U7431, Suzhou Youyi Landi Biotechnology Co., LTD.) Taq SYBR® Green qPCR Premix (Universal) (EG0117M, Jiangsu Bishi Mei Biotechnology Co., LTD.) All-in-One First-Strand Synthesis MasterMix (with dsDNase) was obtained from Jiangsu Bestmate Biotechnology Co., LTD. (item number (EG15133S).

Techniques: Expressing

Journal: Frontiers in Pharmacology

Article Title: Effect of gentiopicroside on endogenous formaldehyde homocysteine–pathway related proteins in rats with non-alcoholic steatohepatitis

doi: 10.3389/fphar.2025.1700101

Figure Lengend Snippet: The mechanism diagram of endogenous formaldehyde participating in one-carbon metabolism leading to homocysteine accumulation and causing NASH. Note: formaldehyde (Endogenous FA); Glutathione (GSH) Mitochondrial aldehyde dehydrogenase 2 (ALDH2); Formate; Tetrahydrofolate (THF); 5, 10-methylenetetrahydrofolate (5, 10-CH2-THF); Methylenetetrahydrofolate reductase (MTHFR); Homocysteine (HCY); S-adenosine homocysteine (SAH); Reactive oxygen species (ROS); Non-alcoholic steatohepatitis (NASH); Methionine Cystathionine -β -synthase (CBS); 5-methyltetrahydrofolate (5-CH3-THF); S-adenosylmethionine (SAM); Methionine adenosyltransferase 1A (MAT1A); S-adenosine homocysteine hydrolase (AHCY).

Article Snippet: ROS, GSH, MDA, SOD, SAH, SAM, GST, CAT, GSH, HCY were obtained from Jiangsu Enzyme Immunoassay Biotechnology Co., LTD. Their item numbers are MM-88564O1, MM-20251R1, MM-2037H1, MM-20387R1, MM-50456H1, MM-0248H1, MM-21254R1, MM-20447R1, MM-20251R1 and MM-50456H1. β-actin antibody (Proteintech, 20536-1-AP); ALDH2 antibody (Proteintech, 15310-1-AP); MAT1A antibody (Proteintech, 12395-1-AP); MTHFR antibody (Proteintech, 26591-1-AP); CBS antibody (Proteintech, 14787-1-AP); AHCY antibody (Proteintech, 10757-2-AP); PVDF membrane with a thickness of 0.45 μm (IPVH00010, Millipore); Universal total RNA extraction reagent (U7431, Suzhou Youyi Landi Biotechnology Co., LTD.) Taq SYBR® Green qPCR Premix (Universal) (EG0117M, Jiangsu Bishi Mei Biotechnology Co., LTD.) All-in-One First-Strand Synthesis MasterMix (with dsDNase) was obtained from Jiangsu Bestmate Biotechnology Co., LTD. (item number (EG15133S).

Techniques:

Journal: Drug Metabolism and Disposition

Article Title: Human Liver Methionine Cycle: MAT1A and GNMT Gene Resequencing, Functional Genomics, and Hepatic Genotype-Phenotype Correlation

doi: 10.1124/dmd.112.046953

Figure Lengend Snippet: Human MAT1A genetic polymorphisms Exons and untranslated regions (UTRs) are numbered relative to the A (nucleotide 1) in the ATG translation initiation codon in exon 1. Negative numbers were assigned to positions 5′ to that location, and positive numbers were assigned to positions 3′. Nucleotides located within introns are numbered on the basis of their distance from the nearest splice junction, with distances from 3′ splice junctions assigned positive numbers and distances from 5′ splice junctions assigned negative numbers. Exon sequences are bold. Polymorphisms identified previously are noted by reference SNP (rs) number.

Article Snippet: Mammalian expression constructs were created for wild-type (WT) MAT1A by subcloning the open reading frame of MAT1A from the Origene clone SC119881 (Origene, Rockville, MD) into pcDNA3.1D/V5-His-TOPO (Invitrogen, Carlsbad, CA) in frame with the V5-His tag, and site-directed mutagenesis was used to create variant allozyme expression constructs.

Techniques: Sequencing

Journal: Drug Metabolism and Disposition

Article Title: Human Liver Methionine Cycle: MAT1A and GNMT Gene Resequencing, Functional Genomics, and Hepatic Genotype-Phenotype Correlation

doi: 10.1124/dmd.112.046953

Figure Lengend Snippet: Human MAT1A and GNMT polymorphisms observed during gene resequencing. The figure shows a schematic representation of the human (A) MAT1A and (B) GNMT gene structures, with arrows indicating the locations of polymorphisms observed during the resequencing studies. Black rectangles represent exons encoding the opening reading frame, and open rectangles represent portions of exons encoding untranslated region sequences. The colors of arrows indicate minor allele frequencies.

Article Snippet: Mammalian expression constructs were created for wild-type (WT) MAT1A by subcloning the open reading frame of MAT1A from the Origene clone SC119881 (Origene, Rockville, MD) into pcDNA3.1D/V5-His-TOPO (Invitrogen, Carlsbad, CA) in frame with the V5-His tag, and site-directed mutagenesis was used to create variant allozyme expression constructs.

Techniques:

Journal: Drug Metabolism and Disposition

Article Title: Human Liver Methionine Cycle: MAT1A and GNMT Gene Resequencing, Functional Genomics, and Hepatic Genotype-Phenotype Correlation

doi: 10.1124/dmd.112.046953

Figure Lengend Snippet: MAT1A allozyme functional genomics Values are mean ± S.E.M. for three independent determinations. The variant allozyme values did not differ significantly ( p > 0.05) from those for the WT allozyme for either apparent K m values or enzyme activity (measured using bacterially expressed protein) or relative protein quantity after expression in COS-1 cells.

Article Snippet: Mammalian expression constructs were created for wild-type (WT) MAT1A by subcloning the open reading frame of MAT1A from the Origene clone SC119881 (Origene, Rockville, MD) into pcDNA3.1D/V5-His-TOPO (Invitrogen, Carlsbad, CA) in frame with the V5-His tag, and site-directed mutagenesis was used to create variant allozyme expression constructs.

Techniques: Functional Assay, Variant Assay, Activity Assay, Expressing

Journal: Drug Metabolism and Disposition

Article Title: Human Liver Methionine Cycle: MAT1A and GNMT Gene Resequencing, Functional Genomics, and Hepatic Genotype-Phenotype Correlation

doi: 10.1124/dmd.112.046953

Figure Lengend Snippet: Human hepatic liver MAT1A and GNMT protein levels. A, MAT1A protein-level frequency distribution. B, GNMT protein-level frequency distribution. C, representative Western blot analysis gel with a, b, c, and d representing individual samples assayed in triplicate.

Article Snippet: Mammalian expression constructs were created for wild-type (WT) MAT1A by subcloning the open reading frame of MAT1A from the Origene clone SC119881 (Origene, Rockville, MD) into pcDNA3.1D/V5-His-TOPO (Invitrogen, Carlsbad, CA) in frame with the V5-His tag, and site-directed mutagenesis was used to create variant allozyme expression constructs.

Techniques: Western Blot

Journal: Drug Metabolism and Disposition

Article Title: Human Liver Methionine Cycle: MAT1A and GNMT Gene Resequencing, Functional Genomics, and Hepatic Genotype-Phenotype Correlation

doi: 10.1124/dmd.112.046953

Figure Lengend Snippet: Genotype-phenotype correlations for human hepatic MAT1A and GNMT. Negative log-transformed p values for single SNP associations with human liver (A) GNMT protein levels and (B) MAT1A protein levels using both genotyped SNPs and SNPs imputed by using 1000 Genomes Project data. Figure 4, A and B, shows SNPs within 20 kb of the 3′ and 5′ ends of the genes and includes only imputed SNPs with imputation quality score Rsq values >0.3. Black circles represent genotyped tag SNPs, and red triangles represent imputed SNPs. *, the threshold for significance after Bonferroni correction for GNMT. The structures of both genes are also shown schematically. C, the relationship between SNP genotypes for the GNMT rs9471976 and rs11752813 SNPs and hepatic GNMT protein is shown. Spearman's rank correlation coefficients (r) and p values are also shown.

Article Snippet: Mammalian expression constructs were created for wild-type (WT) MAT1A by subcloning the open reading frame of MAT1A from the Origene clone SC119881 (Origene, Rockville, MD) into pcDNA3.1D/V5-His-TOPO (Invitrogen, Carlsbad, CA) in frame with the V5-His tag, and site-directed mutagenesis was used to create variant allozyme expression constructs.

Techniques: Transformation Assay

Journal: Drug Metabolism and Disposition

Article Title: Human Liver Methionine Cycle: MAT1A and GNMT Gene Resequencing, Functional Genomics, and Hepatic Genotype-Phenotype Correlation

doi: 10.1124/dmd.112.046953

Figure Lengend Snippet: Spearman correlations and p values for correlations of levels of protein expression measured in 268 hepatic biopsy samples The three enzymes that are expressed primarily in liver, GNMT, MAT1A, and BHMT, are bolded. The p values listed in the table have been corrected for multiple comparisons.

Article Snippet: Mammalian expression constructs were created for wild-type (WT) MAT1A by subcloning the open reading frame of MAT1A from the Origene clone SC119881 (Origene, Rockville, MD) into pcDNA3.1D/V5-His-TOPO (Invitrogen, Carlsbad, CA) in frame with the V5-His tag, and site-directed mutagenesis was used to create variant allozyme expression constructs.

Techniques: Expressing

Journal: Drug Metabolism and Disposition

Article Title: Human Liver Methionine Cycle: MAT1A and GNMT Gene Resequencing, Functional Genomics, and Hepatic Genotype-Phenotype Correlation

doi: 10.1124/dmd.112.046953

Figure Lengend Snippet: Correlations between levels of MAT1A and BHMT protein in adult human liver biopsy samples plotted against log GNMT protein levels in the same samples adjusted for age. A, GNMT versus MAT1A. B, GNMT versus BHMT. Spearman's correlation coefficients, r, as well as p values for associations after correction for multiple comparisons are shown. In both panels, the level of log GNMT protein has been corrected for patient age.

Article Snippet: Mammalian expression constructs were created for wild-type (WT) MAT1A by subcloning the open reading frame of MAT1A from the Origene clone SC119881 (Origene, Rockville, MD) into pcDNA3.1D/V5-His-TOPO (Invitrogen, Carlsbad, CA) in frame with the V5-His tag, and site-directed mutagenesis was used to create variant allozyme expression constructs.

Techniques: