DOI: 10.5061/dryad.n02v6wx6v
title: Therapeutic effects of EGCG in experimentally induced ulcerative colitis in rats via affecting inflammation and apoptosis

description: 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.
description: <strong><em>Animals and treatment
 outlines</em></strong> 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.
 <strong><em>Control
 group:</em></strong> 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. <strong><em>Ucerative
 Colitis group:</em></strong> 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. <strong><em>Ulcerative
 Colitis group treated with genistein:</em></strong> 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.
 <strong><em>Sample
 collection</em></strong> 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.
 <strong><em>Estimation of caspases
 activity</em></strong> 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. <strong><em>Enzyme-linked immunosorbent
 assay (ELISA)</em></strong> 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). <strong><em>Quantitative real-time
 polymerase chain reaction (RT‒PCR)</em></strong>
 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. 
description: # Therapeutic effects of EGCG in experimentally induced ulcerative colitis
 in rats via affecting inflammation and apoptosis
 [https://doi.org/10.5061/dryad.n02v6wx6v](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.