Therapeutic effects of EGCG in experimentally induced ulcerative colitis in rats via affecting inflammation and apoptosis
Data files
Dec 03, 2024 version files 21.20 KB
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README.md
5.68 KB
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Results.xlsx
15.53 KB
Abstract
Ulcerative colitis (UC) is a chronic inflammatory bowel disease that affects the colon. Epigallocatechin gallate (EGCG), a compound found in green tea with antioxidant and anti-inflammatory properties. We aimed to investigate the impact of EGCG on inflammation and apoptotic pathways in UC rats. The study involved inducing UC in rats by administering acetic acid. The UC rats were then treated with EGCG. Colon samples were collected, measured then used to evaluate gene and protein expression of various factors, including NF-κB, TNF-α, SphK, MIP-1α, BCL2, and BAX, as well as the activities of caspase-3/8/9.
README: Therapeutic effects of EGCG in experimentally induced ulcerative colitis in rats via affecting inflammation and apoptosis
https://doi.org/10.5061/dryad.n02v6wx6v
Description of the data and file structure
The uploaded data represented the experiment's results on colons separated from different rat groups used. The data are described in the following table:
Variable | Unit | Description |
---|---|---|
Group | The name of the group. The study consists of three groups: the control group, the ulcerative colitis group (UC), and the ulcerative colitis group treated with EGCG (EGCG). | |
Rat number | Each group consists of twelve rats. The rats are numbered from one to twelve. | |
Colon length | Centimeter (cm) | The length of the colon, which is separated from rats after sacrifice. |
Colon weight | Gram (g) | The weight of the colon, which is separated from the rats after sacrifice. |
SphK gene expression | Relative to control | The relative gene expression of the sphingosine kinase (SphK) in the colon of rats. |
SphK conc | ng/ml | The concentration of the sphingosine kinase (SphK) protein in the colon of rats. |
MIP-1α gene expression | Relative to control | The relative gene expression of the macrophage inflammatory protein-1α (MIP-1α) in the colon of rats. |
MIP-1α concentration | pg/ml | The concentration of the macrophage inflammatory protein-1α (MIP-1α) protein in the colon of rats. |
NFκB gene expression | Relative to control | The relative gene expression of the nuclear factor κB (NFκB) in the colon of rats. |
TNF-α gene expression | Relative to control | The relative gene expression of the tumor necrosis factor-α (TNF-α) in the colon of rats. |
TNF-α concentration | pg/ml | The concentration of the tumor necrosis factor-α (TNF-α) protein in the colon of rats. |
BCL2 gene expression | Relative to control | The relative gene expression of the B-cell lymphoma-2 (BCL2) in the colon of rats. |
BCL2 concentration | ng/l | The concentration of the B-cell lymphoma 2 (BCL2) protein in the colon of rats. |
BAX gene expression | Relative to control | The relative gene expression of the BCL2 Associated X (BAX) in the colon of rats. |
BAX concentration pg/ml | pg/ml | The concentration of the BCL2 Associated X (BAX) protein in the colon of rats. |
Caspase-3 OD (405 nm) | The value of absorbance of the reagent mixture at 405 nm. | |
Caspase-8 OD (405 nm) | The value of absorbance of the reagent mixture at 405 nm. | |
Caspase-9 OD (405 nm) | The value of absorbance of the reagent mixture at 405 nm. |
Files and variables
File: Results.xlsx
Description: The file contains all the results of the measurements that was collected in the animal study.
Variables
- B-cell lymphoma 2 (BCL2)
- BCL2 Associated X (BAX)
- Caspase-3/8/9
- epigallocatechin gallate (EGCG)
- macrophage inflammatory protein-1α (MIP-1α)
- nuclear factor (NF)κB
- sphingosine kinase (SphK)
- tumor necrosis factor (TNF)-α
- ulcerative colitis (UC).
Code/software
The files are made using the Excel program by Microsoft.
Access information
Other publicly accessible locations of the data:
- There is no other publicly accessible location of the data.
Data was derived from the following sources:
- Data are the results of our experiments in rats.
Methods
Animals and treatment outlines
In this study, 36 rats belonging to the Sprague-Dawley species and weighing between 180 and 200 grams were employed. The rats were divided into three groups, each consisting of twelve rats.
Control group: Rats were subjected to mild anesthesia using ether followed by the administration of 2 ml of saline solution into the colon using a soft pediatric lubricated catheter. The rats were positioned horizontally for two minutes to prevent the saline from draining. Subsequently, they received a daily oral dose of 0.5% (w/v) methylcellulose and 2% (v/v) Tween 80, both dissolved in deionized water, for two weeks.
Ucerative Colitis group: The rats were anesthetized with ether and subsequently administered 2 ml of 4% acetic acid via a gentle pediatric catheter into their colon. Following the administration, the rats were positioned horizontally for two minutes to promote the retention of the acetic acid.
Ulcerative Colitis group treated with genistein: UC was induced utilizing the previously outlined methodology. Subsequently, the rats received an oral administration of 20 mg/kg of EGCG (Sigma-Aldrich, Inc, St. Louis, MO, United States) once daily for two weeks.
Sample collection
After the animals were euthanized, the entire colon was excised, measured, and weighed. A segment was homogenized in a ten-fold cold sodium-potassium phosphate buffer (0.01 M, pH 7.4) containing 1.15% KCl. The resulting solution was then subjected to rapid freezing at -80°C.
Estimation of caspases activity
The enzyme activities of caspase-3, -8, and -9 were evaluated using commercially available kits (Abcam, Cambridge, MA, USA). The Caspase assay protocol is based on the formation of the chromophore p-nitroaniline by cleavage from the labeled substrate DEVD-pNA. The p-NA can be quantied using a spectrophotometer or a microtiter plate reader reading absorbance at 405 nm.
Enzyme-linked immunosorbent assay (ELISA)
We utilized commercially available ELISA kits to evaluate the levels of SphK, macrophage inflammatory protein (MIP)-1α (CCL3), B-cell lymphoma 2 (BCL2), and BCL2 Associated X (BAX), as well as TNF-α under the provided instructions from the manufacturer (MyBioSource, Inc., San Diego, CA, USA).
Quantitative real-time polymerase chain reaction (RT‒PCR)
The mRNA levels of SphK, MIP-1α, BCL2, BAX, NFκB, and TNF-α in rat colon lysates were analyzed using established protocols. β-actin was used as the internal reference and housekeeping gene.