Croton Oil-induced Hemorrhoidal Rats: Evaluation of Anti-inflammatory
Mediated Anti-hemorrhoidal Activity of Tridax procumbens Whole Plant
Authors: Nallajerla Sai Krishna , Ganta Suhasin 
Published: February 2023

Background:
Hemorrhoids are anorectal disorders characterized by dilation of rectal
blood vessels, inflammation, and prolapse of the anal tissue. The disease affects both men and
women equally. In consequence, the increasing prevalence of the disease needs potential agents
with efficiency and low cost to support the treatment.

Objective:
The treatment of hemorrhoids in modern medicine is still found to be in its infancy, as
there is no precise treatment for the specific disease. Tridax procumbens Linn (T. procumbens) belongs
to the family Asteraceae. From the literature, the plant was found to have a traditional use for
the treatment of hemorrhoids. Hence, the present research was focused on evaluating the antiinflammatory
mediated anti-hemorrhoidal potential of ethanolic whole plant extract of T. procumbens
(EWETP) in croton oil-induced hemorrhoidal rats.

Methods:
An in vitro study was conducted to evaluate the percentage inhibition of protein (egg albumin)
denaturation by Tridax procumbens. This study determined the initial protective role of plant
extracts against inflammatory proteins. An in vivo, anti-inflammatory-mediated anti-hemorrhoidal
study was carried out on T. procumbens in various groups of croton oil-induced hemorrhoidal rats.
Wistar albino rats were selected for the present research, and hemorrhoids were induced in experimental
animals using a croton oil preparation containing 6% croton oil, deionised water, pyridine,
and diethyl ether. Pilex ointment was taken as a reference drug in this study. Inflammation seems to
be a major pathway for the progression of hemorrhoids. Hence, in the present research, Evans blue
(EB) extravasation technique was applied to quantify inflammatory proteins. On the last day of the
study, blood samples were drawn from experimental animals to analyse serum and blood for TNF-
α, IL-6, and the percentage of neutrophils count. Recto anal coefficient was calculated to measure
croton oil-induced rectal inflammation in animals. Histopathological studies were carried out separately
on a second set of animals to identify the protective role of T.procumbens on rectal tissue cell
histology. Molecular docking studies were carried out to rule out the possible interaction of plant
phytoconstituents with the COX-2 enzyme.

Results:
Results showed that intra rectal application of croton oil preparation in albino rats developed
hemorrhoids by elevating serum TNF-α and IL-6 in positive control group animals compared
to normal group rats. Treatment of albino rats with T. procumbens at doses of 200 mg/kg and 400
mg/kg in groups IV and V has shown a significant reduction in serum TNF-α and IL-6. Furthermore,
in the study, T. procumbens exhibited a significant dose-dependent reduction of EB dye extravasation
in study animals. The study observations also revealed the inhibitory effect of plant extract
on the blood percentage of neutrophils count and recto anal coefficient compared to the positive
control group rats. Histopathological studies conducted on rectoanal tissues showed the presence
of minimal rectal tissue changes in plant extract-treated group animals compared to positive
control group rats. Molecular docking studies explored the possible interaction of phenolic compounds
of plant extract with the COX enzyme.

Conclusion:
It was concluded that Tridax procumbens had a protective role against inflammatory
mediators in hemorrhoids. In hemorrhoidal rats, ethanolic leaf extract was found to reduce the
plasma percentage of neutrophils and other inflammatory cytokines, TNF-α, and IL-6. It could be
used as a therapeutic anti-inflammatory mediated anti-hemorrhoidal agent.