Data for: D-limonene mitigates doxorubicin-induced cardiotoxicity
Data files
Nov 06, 2026 version files 5.61 MB
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1._Blots.pdf
307.30 KB
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ASC.jpg
582.73 KB
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Cleaved_caspase_1.jpg
697.46 KB
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GAPDH.jpg
341.04 KB
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GAPDHh.jpg
279.94 KB
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Gasdermin-D_N.jpg
436.16 KB
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LC3-I__LC3-II.jpg
462.58 KB
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NF-kB.jpg
500.54 KB
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NLRP3.jpg
486.36 KB
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P62.jpg
572.45 KB
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Parkin.jpg
449.61 KB
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Pink1.jpg
446.18 KB
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Raw_data2.xlsx
33.42 KB
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README.md
16.13 KB
Abstract
This study investigates the cardioprotective effects of a monocyclic monoterpene (D-Lim) against doxorubicin-induced myocardial injury in rats, with a particular focus on mitochondrial quality control and inflammasome-mediated pyroptosis. Our findings demonstrate that D-Lim significantly improves cardiac function and structure by restoring mitochondrial function and mitophagy proteins and suppressing NLRP3 inflammasome activation. These results provide novel mechanistic insight into mitochondrial–inflammatory interaction in chemotherapy-induced cardiotoxicity and highlight D-Lim as a potential adjuvant cardioprotective strategy.
Dataset DOI: 10.5061/dryad.x95x69q2g
Description of the data and file structure
This dataset contains the raw experimental data and associated files used in the manuscript entitled D-Limonene Mitigates Doxorubicin-Induced Cardiotoxicity Through Mitochondrial and Inflammasome Modulation. The dataset includes PDF files containing the complete collection of Western blot images, as well as individual image files for each analyzed protein. Additionally, an Excel workbook provides the experimental data, including hemodynamic parameters, histological index, biochemical measurements, ELISA results, body weights, mitochondrial parameters, and individual Western blot quantification data.
Files and variables
File: 1._Blots.pdf
Description: Complete collection of Western blot images
File: ASC.jpg
Description: Western blot image for apoptosis-associated speck-like protein containing a CARD
File: Cleaved_caspase_1.jpg
Description:
Western blot image for cleaved caspase 1
File: GAPDH.jpg
Description:
Western blot image for glyceraldehyde 3-phosphate dehydrogenase
File: GAPDHh.jpg
Description:
Western blot image for glyceraldehyde 3-phosphate dehydrogenase
File: Gasdermin-D_N.jpg
Description:
Western blot image for gasdermin-D N terminal fragment
File: LC3-I__LC3-II.jpg
Description:
Western blot image for microtubule-associated proteins light chain 3, functional forms I and II
File: NLRP3.jpg
Description:
Western blot image for NLR family pyrin domain containing 3
File: Parkin.jpg
Description:
Western blot image for Parkin
File: NF-kB.jpg
Description:
Western blot image for nuclear factor kappa-light-chain-enhancer of activated B cells
File: P62.jpg
Description:
Western blot image for sequestosome-1
File: Pink1.jpg
Description:
Western blot image for PTEN-induced kinase 1
File: Raw_data2.xlsx
Description: raw data including including hemodynamic parameters, histological index, biochemical measurements, ELISA results, body weights, and individual Western blot quantification data.
Group Allocation and Data Structure in Excel File
- Animals were randomly allocated into five experimental groups. In the Excel data sheets, each row represents an individual animal/sample, identified by its sample number in the first column, followed by five columns indicating the experimental group allocation. Column A contains the sample numbers, while Columns B–F represent the five experimental groups (Groups 1–5: Control, DOX, DOX+D-lim, DOX+Mdv, and DOX+D-lim+Mdv, respectively). The individual sample values for each parameter are arranged in rows beneath the corresponding group column. Each experimental group includes six biological samples (n = 6), except for Western blot analyses, which include three biological samples per group. The animal groups in the supplementary data sheet comprise two groups: Control and D-limonene (Lim), with six biological samples in each group (n = 6/group).
Description of variables in excel file:
| Sheet in Excel file | Variable Name | Description | Units | Method of measurement |
|---|---|---|---|---|
| Sheet 1 | LVSP | Levels of left ventricular systolic pressure | mmHg | Invasive left ventricular pressure measurement using a pressure transducer and data acquisition system |
| Sheet 2 | LVEDP | Levels of left ventricular end-diastolic pressure | mmHg | Invasive left ventricular pressure measurement using a pressure transducer and data acquisition system |
| Sheet 3 | LVSP | Levels of left ventricular systolic pressure | mmHg | Invasive left ventricular pressure measurement using a pressure transducer and data acquisition system |
| Sheet 4 | cTnI | Levels of cardiac troponin I, a biomarker of myocardial injury | pg/ml | Enzyme-linked immunosorbent assay (ELISA) |
| Sheet 5 | Histology index | Semi-quantitative score representing the severity of myocardial histopathological alterations | Score (dimensionless) | Histological examination of hematoxylin and eosin (H&E)-stained cardiac tissue sections using a predefined scoring system |
| Sheet 6 | NLRP3 | Levels of NOD-like receptor family pyrin domain-containing 3 protein expression | pg/mg protein | ELISA |
| Sheet 7 | LDH | Levels of lactate dehydrogenase activity, an indicator of cellular injury | U/L | Enzymatic colorimetric assay |
| Sheet 8 | IL-1β | Levels of interleukin-1 beta, a pro-inflammatory cytokine | pg/mg protein | ELISA |
| Sheet 9 | TNF-α | Levels of tumor necrosis factor-alpha, a pro-inflammatory cytokine | pg/mg protein | ELISA |
| Sheet 10 | ASC | Levels of apoptosis-associated speck-like protein containing a CARD, an inflammasome adaptor protein | fold changes | Western blot analysis |
| Sheet 11 | C. caspase 1 | Levels of cleaved caspase-1, the activated form of caspase-1 involved in inflammasome signaling | fold changes | Western blot analysis |
| Sheet 12 | GasderminD | Levels of gasdermin D, a protein involved in pyroptosis; its cleaved N-terminal fragment | fold changes | Western blot analysis |
| Sheet 13 | NF-κB | Levels of nuclear factor kappa B, a transcription factor involved in inflammatory signaling | fold changes | Western blot analysis |
| Sheet 14 | NLRP3 | Levels of NLR family pyrin domain-containing 3, an inflammasome component | fold changes | Western blot analysis |
| Sheet 15 | Pink 1 | Levels of PTEN-induced kinase 1, a mitochondrial quality-control protein involved in mitophagy | fold changes | Western blot analysis |
| Sheet 16 | Parkin | Levels of E3 ubiquitin-protein ligase Parkin, involved in mitochondrial quality control and mitophagy | fold changes | Western blot analysis |
| Sheet 17 | LC3-II | Levels of microtubule-associated protein 1A/1B-light chain 3-II, a marker associated with autophagosomes | fold changes | Western blot analysis |
| Sheet 18 | P62 | Levels of sequestosome-1 (SQSTM1/p62), an autophagy-related cargo receptor | fold changes | Western blot analysis |
| Sheet 19 | Mitochondrial ROS | Levels of reactive oxygen species generated within mitochondria | FI (Fluorescent intensity) | Fluorescence-based assay using a mitochondria-targeted ROS probe |
| Sheet 20 | Mitochondrial membrane potential | Levels of mitochondrial inner membrane potential, reflecting mitochondrial functional integrity | FI (Fluorescent intensity) | Fluorescence-based assay using JC-1 dye |
| Sheet 21 | ATP | Levels of adenosine triphosphate content, reflecting cellular energy status | μmol/ mg protein | Luciferase-based ATP quantification assay |
| Sheet 22 | Initial Body Weight | Levels of body weight of each animal at the beginning of the experimental period | g | Measurement using a calibrated digital weighing scale |
| Sheet 23 | Last Body Weight | Levels of body weight of each animal at the end of the experimental period | g | Measurement using a calibrated digital weighing scale |
| Sheet 24 | Body Weight Gain | Difference between final and initial body weights levels | g | Calculated as final body weight minus initial body weight |
| Sheet 25 | Supplementary Fig. S1 | Levels of parameters including: LVEDP, LVSP, LVDP, cTnI, NLRP3, LDH, IL-1β, TNF-α, mitochondrial ROS, mitochondrial membrane potential, ATP | As above for each parameter | As above for each parameter, including invasive pressure assessment technique, ELISA, colorimetric assay, Mitochondrial probe and fluorescence- and luciferase-based assay |
Experimental Design
The five experimental groups were as follows:
1. Control: Rats received normal saline intraperitoneally (i.p.) for 12 days.
2. DOX: Rats received doxorubicin (2.5 mg/kg, i.p.) every other day for 12 days to induce cumulative and progressive cardiotoxicity.
3. DOX + D-lim: Rats received D-limonene (50 mg/kg/day, orally) for 19 days, starting 7 days before the first doxorubicin injection and continuing throughout the experimental period. Doxorubicin was administered using the same regimen as in the DOX group.
4. DOX + Mdv: Rats received Mdivi-1 (1.2 mg/kg/day, i.p.), a pharmacological inhibitor of mitochondrial fission, for 19 days, starting 7 days before the first doxorubicin injection and continuing throughout the experimental period. Doxorubicin was administered using the same regimen as in the DOX group.
5. DOX + D-lim + Mdv: Rats received combined treatment with D-limonene (50 mg/kg/day, orally) and Mdivi-1 (1.2 mg/kg/day, i.p.) for 19 days, starting 7 days before the first doxorubicin injection and continuing throughout the experimental period. Doxorubicin was administered using the same regimen as in the DOX group.
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Additional notes for data interpretation
- DOX: Doxorubicin.
- D-lim or Lim: D-limonene.
- Mdv: Mdivi-1.
- i.p.: Intraperitoneal administration.
Code/software
Acrobat PDF reader, Microsoft Excel and any imaging software and tools