Interleukin-6 Mediates Myocardial Fibrosis, Concentric Hypertrophy and Diastolic Dysfunction in Rats methods
Aim. Evidence-backed execution summary for Interleukin-6 Mediates Myocardial Fibrosis, Concentric Hypertrophy and Diastolic Dysfunction in Rats methods from Interleukin-6 Mediates Myocardial Fibrosis, Concentric Hypertrophy and Diastolic Dysfunction in Rats.
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rat
Subject model for the experiment.
- Use
- confirm full cohort details in the source paper
Materials and Methods
reagent used in the protocol.
- Use
- IL-6 (Invitrogen, USA) was administered at a rate of 2.5 µg·kg -1 ·hr -1 for 7 days using osmotic minipumps (Alzet model 2001, Durect Corporation, Cupertino CA) implanted in the peritoneal cavity. The lyophilized IL-6 was dissolved in a vehicle solution containing 10% rat albumin and. At the experimen...
Determination of Cardiomyocyte Size
reagent used in the protocol.
- Use
- Cardiomyocytes from the LV of vehicle (n=6) and IL-6 infused (n=8) rats were isolated using a modified KOH procedure initially described by Gerdes et al. to assess the extent of cellular remodeling. Briefly, formalin fixed tissue from a mid-level transmural section of the LV trimmed into 1 mm 3 pieces was placed in...
Determination of Cardiomyocyte Size
Cardiomyocytes from the LV of vehicle (n=6) and IL-6 infused (n=8) rats were isolated using a modified KOH procedure initially described by Gerdes et al. to assess the extent of cellular remodeling. Briefly, formalin fixed tissue from a mid-level transmural section of the LV trimmed into 1 mm 3 pieces was placed in...
- Use
- Cardiomyocytes from the LV of vehicle (n=6) and IL-6 infused (n=8) rats were isolated using a modified KOH procedure initially described by Gerdes et al. to assess the extent of cellular remodeling. Briefly, formalin fixed tissue from a mid-level transmural section of the LV trimmed into 1 mm 3 pieces was placed in...
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Materials and Methods
Adult male Sprague-Dawley rats weighing between 250 and 300 g were housed under standard environmental condition and maintained on commercial rat chow and tap water ad libitum. This investigation conformed with the recommendations in the Guide for the Care and Use of Laboratory Animals published by the US National Institutes of Health (NIH Publication No. 85-23, revised 1996), and the experimental protocol had Institutional Animal Care and Use Committee approval. Anesthesia for the non-terminal surgical procedure was achieved by inhalation of isoflurane (2%). Analgesia was achieved by administration of buprenorphine HCL (0.025 mg·kg -1, SQ) to the rats at the time of surgery. At the experimental endpoint the rats were anesthetized with pentobarbital (70 mg·kg -1, IP) and euthanized by removal of the heart.
Materials and Methods
IL-6 (Invitrogen, USA) was administered at a rate of 2.5 µg·kg -1 ·hr -1 for 7 days using osmotic minipumps (Alzet model 2001, Durect Corporation, Cupertino CA) implanted in the peritoneal cavity. The lyophilized IL-6 was dissolved in a vehicle solution containing 10% rat albumin and. At the experimental endpoint, blood pressure and body weights were measured and left ventricular (LV) function assessed.
Determination of Cardiomyocyte Size
Cardiomyocytes from the LV of vehicle (n=6) and IL-6 infused (n=8) rats were isolated using a modified KOH procedure initially described by Gerdes et al. to assess the extent of cellular remodeling. Briefly, formalin fixed tissue from a mid-level transmural section of the LV trimmed into 1 mm 3 pieces was placed in a 0.1M KOH solution at room temperature for 24 hours. The tissue was then transferred to a 0.1 M PBS solution and continuously agitated for 10 minutes. Cardiac myocytes thus obtained were purified using a 10% Ficoll density gradient. Finally, the cells were resuspended in 10% buffered formalin and transferred to a glass microscope slide, using a cytospin apparatus, and stained with 1.5% gallocyanin and 5% chromium potassium sulfate dodecahydrate. The length and width of 50 rod-shaped cardiomyocytes were measured for each individual heart using ImagePro® Plus software (...
Measurement outputs
What raw and processed outputs should exist?
The effect of IL-6 infusion on LV collagen volume fraction is presented in. A greater than 3-fold increase in interstitial myocardial collagen was observed in the IL-6 infused...
- Raw artifact
- Per-sample or per-animal endpoint measurements collected during the experiment
- Processed artifact
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- Summary statistics and between-group or across-timepoint comparisons
Analysis plan
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Acquisition
Collect raw experimental outputs with enough metadata to preserve sample identity, condition, and timing.
inferred from protocolPreprocessing / cleaning
The effect of IL-6 infusion on LV collagen volume fraction is presented in.
from paperScoring or quantification
Quantify the primary readouts for this experiment: The effect of IL-6 infusion on LV collagen volume fraction is presented in. A greater than 3-fold increase in interstitial myocardial collagen was observed in the IL-6 infused....
from paperStatistical comparison
The effect of IL-6 infusion on LV collagen volume fraction is presented in. A greater than 3-fold increase in interstitial myocardial collagen was observed in the IL-6 infused...
from paperReporting output
Report representative outputs alongside summary comparisons for The effect of IL-6 infusion on LV collagen volume fraction is presented in. A greater than 3-fold increase in interstitial myocardial collagen was observed in the IL-6 infused....
inferred from protocolStructured statistical methods
The effect of IL-6 infusion on LV collagen volume fraction is presented in. A greater than 3-fold increase in interstitial myocardial collagen was observed in the IL-6 infused...
source structuredSource and audit
What supports the facts on this page?
Evidence quotes (3)
Adult male Sprague-Dawley rats weighing between 250 and 300 g were housed under standard environmental condition and maintained on commercial rat chow and tap water ad libitum. This investigation conformed with the recommendations in the Guide for the Care and Use of Laboratory Animals published by the US National Institutes of Health (NIH Publication No. 85-23, revised 1996), and the experimental protocol had Institutional Animal Care and Use Committee approval. Anesthesia for the non-terminal surgical procedure was achieved by inhalation of isoflurane (2%). Analgesia was achieved by administration of buprenorphine HCL (0.025 mg·kg -1, SQ) to the rats at the time of surgery. At the experimental endpoint the rats were anesthetized with pentobarbital (70 mg·kg -1, IP) and euthanized by removal of the heart.
IL-6 (Invitrogen, USA) was administered at a rate of 2.5 µg·kg -1 ·hr -1 for 7 days using osmotic minipumps (Alzet model 2001, Durect Corporation, Cupertino CA) implanted in the peritoneal cavity. The lyophilized IL-6 was dissolved in a vehicle solution containing 10% rat albumin and. At the experimental endpoint, blood pressure and body weights were measured and left ventricular (LV) function assessed.
Cardiomyocytes from the LV of vehicle (n=6) and IL-6 infused (n=8) rats were isolated using a modified KOH procedure initially described by Gerdes et al. to assess the extent of cellular remodeling. Briefly, formalin fixed tissue from a mid-level transmural section of the LV trimmed into 1 mm 3 pieces was placed in a 0.1M KOH solution at room temperature for 24 hours. The tissue was then transferred to a 0.1 M PBS solution and continuously agitated for 10 minutes. Cardiac myocytes thus obtained were purified using a 10% Ficoll density gradient. Finally, the cells were resuspended in 10% buffered formalin and transferred to a glass microscope slide, using a cytospin apparatus, and stained with 1.5% gallocyanin and 5% chromium potassium sulfate dodecahydrate. The length and width of 50 rod-shaped cardiomyocytes were measured for each individual heart using ImagePro® Plus software (Media Cybernetics, LP, Silver Springs, MD).
Machine-readable layer
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