Journal: Current Issues in Molecular Biology

Article Title: Identification and Validation of MTFP1 as a Mitochondrial Target Restoring Dynamics and ECM Remodeling in Acute Myocardial Infarction

doi: 10.3390/cimb48030293

Figure Lengend Snippet: Flowchart of analysis. LASSO, Least Absolute Shrinkage and Selection Operator; ROC, receiver operating characteristics; KEGG, Kyoto Encyclopedia of Genes and Genomes; GO, Gene Ontology; ANN, artificial neural network; GSEA, Gene Set Enrichment Analysis; IHC, immunohistochemistry; RT-QPCR, quantitative real-time polymerase chain reaction; AUC, the area below the curve; MTFP1, mitochondrial fission process 1; DNAJC28, DnaJ heat shock protein family (Hsp40) member C2.

Article Snippet: Following a 30 min room temperature block with 3% bovine serum albumin (BSA), the sections were incubated overnight at 4 °C with primary antibodies, including MTFP1 antibody (proteintech, Cat#:14257-1-AP; dilution ratio 1:200).

Techniques: Selection, Immunohistochemistry, Quantitative RT-PCR, Real-time Polymerase Chain Reaction

Journal: Current Issues in Molecular Biology

Article Title: Identification and Validation of MTFP1 as a Mitochondrial Target Restoring Dynamics and ECM Remodeling in Acute Myocardial Infarction

doi: 10.3390/cimb48030293

Figure Lengend Snippet: GeneMANIA network analysis and single-gene GSEA enrichment analysis. ( A ) GeneMANIA interaction network of key genes. ( B ) GSEA enrichment of DNAJC28. ( C ) GSEA enrichment of MTFP1. The red area (left region) represents the region where genes positively correlated with the target phenotype are enriched. At the left side of the ranked gene list (the end with smaller rank values), genes highly positively correlated with AMI are concentrated. If the peak of the enrichment score curve for a gene set falls on the left side, it indicates that the gene set is primarily enriched among positively correlated genes, potentially promoting the disease process. The blue area (right region) represents the region where genes negatively correlated with the target phenotype are enriched. At the right side of the ranked gene list (the end with larger rank values), genes highly negatively correlated with AMI are concentrated. If the peak of the enrichment score curve for a gene set falls on the right side, it indicates that the gene set is primarily enriched among negatively correlated genes, potentially inhibiting the disease process.

Article Snippet: Following a 30 min room temperature block with 3% bovine serum albumin (BSA), the sections were incubated overnight at 4 °C with primary antibodies, including MTFP1 antibody (proteintech, Cat#:14257-1-AP; dilution ratio 1:200).

Techniques:

Journal: Current Issues in Molecular Biology

Article Title: Identification and Validation of MTFP1 as a Mitochondrial Target Restoring Dynamics and ECM Remodeling in Acute Myocardial Infarction

doi: 10.3390/cimb48030293

Figure Lengend Snippet: Dual network analyses and five targeted drugs for biomarkers. ( A ) Transcription factor-gene regulatory network, showing predicted upstream TFs targeting MTFP1 and DNAJC28 ; each green node represents a TF, with purple edges denoting regulatory interactions. ( B ) Competing endogenous RNA (ceRNA) regulatory network for MTFP1 and DNAJC28 . Orange nodes represent lncRNAs, green nodes represent miRNAs, and purple edges denote miRNA–mRNA and lncRNA–miRNA interactions. Key interactions include mmu-miR-155-5p and mmu-miR-124-3p co-regulating both target genes. A Vina score lower than −5 kcal/mol indicates favorable binding affinity. Notably, epigallocatechin gallate showed the strongest docking affinity with both targets (−7.1/−7.6 kcal/mol). ( C , D ) Predicted docking conformation of epigallocatechin gallate (EGCG) within the MTFP1 and DNAJC28 binding pocket. Electrostatic surface rendering highlights hydrogen bond interactions and hydrophobic contacts.

Article Snippet: Following a 30 min room temperature block with 3% bovine serum albumin (BSA), the sections were incubated overnight at 4 °C with primary antibodies, including MTFP1 antibody (proteintech, Cat#:14257-1-AP; dilution ratio 1:200).

Techniques: Binding Assay

Journal: Current Issues in Molecular Biology

Article Title: Identification and Validation of MTFP1 as a Mitochondrial Target Restoring Dynamics and ECM Remodeling in Acute Myocardial Infarction

doi: 10.3390/cimb48030293

Figure Lengend Snippet: Expression profiles of biomarkers MTFP1 and DNAJC28 in vivo and in vitro models. ( A ) mRNA expression levels of MTFP1 in different cardiac regions of C57/BL6 male mice randomly assigned to sham or MI groups (n = 3). ( B ) Temporal mRNA expression patterns of MTFP1 at different post-MI timepoints. ( C ) mRNA expression levels of DNAJC28 in different cardiac regions. ( D ) Temporal mRNA expression patterns of DNAJC28 at different post-MI timepoints. ( E , F ) Protein expression of MTFP1 in sham and MI groups. ( G , H ) MTFP1 protein expression in cardiomyocytes under normoxia vs. hypoxia at indicated durations. ( I ) MTFP1 mRNA expression in cardiomyocytes during hypoxic exposure. ( J ) Representative images of HE staining (scale bar = 200 μm). Masson’s trichrome (scale bar = 200 μm). Picrosirius red staining (scale bar = 200 μm). MTFP1 immunohistochemistry (scale bar = 200 μm) in sham vs. MI hearts. ( K , L ) Quantification of Collagen Content by Masson and Sirius Red Staining expressed as a percentage. ( M ) Mean optical density of MTFP1 in sham operation group and myocardial infarction group. All experiments were conducted in triplicate with three independent biological replicates per group. Data are presented as mean ± SEM. Statistical analysis was performed using unpaired two-tailed Student’s t -test for two-group comparisons and one-way ANOVA followed by Tukey’s post hoc test for multiple-group comparisons. Significance levels were defined as: p < 0.05 (*), p < 0.01 (**), p < 0.001 (***), and p < 0.0001 (****).

Article Snippet: Following a 30 min room temperature block with 3% bovine serum albumin (BSA), the sections were incubated overnight at 4 °C with primary antibodies, including MTFP1 antibody (proteintech, Cat#:14257-1-AP; dilution ratio 1:200).

Techniques: Expressing, In Vivo, In Vitro, Staining, Immunohistochemistry, Two Tailed Test

Journal: Current Issues in Molecular Biology

Article Title: Identification and Validation of MTFP1 as a Mitochondrial Target Restoring Dynamics and ECM Remodeling in Acute Myocardial Infarction

doi: 10.3390/cimb48030293

Figure Lengend Snippet: MTFP1 overexpression ameliorates cardiac function, reduces infarct size, and mitigates post-MI fibrosis. ( A , B ) Validation of MTFP1 overexpression. C57/BL6 male mice were randomly allocated into: control group (n = 3), GFP group (n = 3), MTFP1 overexpression group (n = 3). MTFP1 expression levels were quantified to confirm successful overexpression. ( C – G ) Echocardiographic Assessment of Cardiac Function. Mice were randomized into: sham group (n = 3), MI group (n = 3), MI + GFP group (n = 3), MI + MTFP1 group (n = 3). Parameters evaluated: Ejection fraction (EF%), fractional shortening (FS%), left ventricular end-diastolic diameter (LVEDD; mm), left ventricular end-systolic diameter (LVESD; mm). ( H , I ) Infarct size quantification by TTC staining. Experimental groups: Sham (n = 3), MI (n = 3), MI + MTFP1 (n = 3). ( J – L ) Histopathological evaluation. Groups analyzed: Sham (n = 3), MI (n = 3), MI + GFP (n = 3), MI + MTFP1 (n = 3). Staining performed: HE staining (scale bar = 200 μm), Masson’s trichrome (scale bar = 200 μm), Picrosirius red (scale bar = 200 μm). All experiments were independently repeated three times. All experiments were conducted in triplicate with three independent biological replicates per group. Data are presented as mean ± SEM. Statistical analysis was performed using one-way ANOVA followed by Tukey’s post hoc test for multiple-group comparisons. Significance levels were defined as: p < 0.05 (*), p < 0.01 (**), p < 0.001 (***), and p < 0.0001 (****).

Article Snippet: Following a 30 min room temperature block with 3% bovine serum albumin (BSA), the sections were incubated overnight at 4 °C with primary antibodies, including MTFP1 antibody (proteintech, Cat#:14257-1-AP; dilution ratio 1:200).

Techniques: Over Expression, Biomarker Discovery, Control, Expressing, Staining

Journal: Current Issues in Molecular Biology

Article Title: Identification and Validation of MTFP1 as a Mitochondrial Target Restoring Dynamics and ECM Remodeling in Acute Myocardial Infarction

doi: 10.3390/cimb48030293

Figure Lengend Snippet: MTFP1 restores mitochondrial fission–fusion balance and ECM remodeling via p-DRP1/MMP9/TIMP1 axis in hypoxia-induced cardiomyocytes. ( A , B ) Western blot analysis of myocardial tissue from MI and MI + MTFP1 overexpression (OE) groups assessing MTFP1, DRP1, p-DRP1 (Ser616), MMP9, and TIMP1 expression. Quantification of protein expression is shown (n = 3/group). ( C , D ) Gelatin zymography of myocardial tissue showing pro-MMP9 and active MMP9 levels, with corresponding quantification (n = 3/group). ( E , F ) Western blot analysis of fibrosis-associated proteins COL1A1 and α-SMA in myocardial tissue and corresponding quantification (n = 3/group). ( G , H ) ROS levels in hypoxia-induced H9C2 cell detected by DCFH-DA fluorescence staining and quantitative fluorescence intensity analysis (n = 3/group; scale bar = 200 μm). ( I ) Schematic illustration of the proposed mechanism: MTFP1 overexpression restores mitochondrial dynamics and ECM remodeling by regulating the p-DRP1/MMP9/TIMP1 signaling axis, thereby attenuating fibrosis and oxidative stress while improving cardiac function. All experiments were conducted in triplicate with three independent biological replicates per group. Data are presented as mean ± SEM. Statistical analysis was performed using unpaired two-tailed Student’s t -test for two-group comparisons and one-way ANOVA followed by Tukey’s post hoc test for multiple-group comparisons. Significance levels were defined as: p < 0.01 (**), p < 0.001 (***), and p < 0.0001 (****).

Article Snippet: Following a 30 min room temperature block with 3% bovine serum albumin (BSA), the sections were incubated overnight at 4 °C with primary antibodies, including MTFP1 antibody (proteintech, Cat#:14257-1-AP; dilution ratio 1:200).

Techniques: Western Blot, Over Expression, Expressing, Zymography, Fluorescence, Staining, Two Tailed Test