Basic Study
Copyright ©The Author(s) 2020. Published by Baishideng Publishing Group Inc. All rights reserved.
World J Gastroenterol. Apr 14, 2020; 26(14): 1601-1612
Published online Apr 14, 2020. doi: 10.3748/wjg.v26.i14.1601
Silymarin, boswellic acid and curcumin enriched dietetic formulation reduces the growth of inherited intestinal polyps in an animal model
Bruna Girardi, Maria Pricci, Floriana Giorgio, Mariano Piazzolla, Andrea Iannone, Giuseppe Losurdo, Mariabeatrice Principi, Michele Barone, Enzo Ierardi, Alfredo Di Leo
Bruna Girardi, Maria Pricci, Floriana Giorgio, THD SpA, Correggio 42015, Italy
Mariano Piazzolla, Andrea Iannone, Giuseppe Losurdo, Mariabeatrice Principi, Michele Barone, Enzo Ierardi, Alfredo Di Leo, Gastroenterology Section, Department of Emergency and Organ Transplantation, University of Bari, Bari 70124, Italy
Author contributions: Di Leo A, Ierardi E and Barone M planned the study; Girardi B, Pricci M, Giorgio F, Piazzolla M took care of mice; Girardi B, Pricci M, Giorgio F, Piazzolla M, Losurdo G, Iannone A collected the data; Girardi B, Pricci M, Giorgio F, Piazzolla M performed experimental analysis; Ierardi E and Girardi B performed histological analysis; Iannone A and Losurdo G performed statistical analysis; Girardi B and Ierardi E wrote the paper; All Authors read and approved the final version.
Institutional review board statement: The study protocol was approved by the University of Bari Ethics committee (protocol number 6/12).
Institutional animal care and use committee statement: The study protocol was approved by the University of Bari Committee for Animal Experimentation (protocol number 6/12).
Conflict-of-interest statement: Alfredo Di Leo is an advisory board member of THD S.p.a. Floriana Giorgio, Bruna Girardi and Maria Pricci are employees of THD S.p.a. All the other authors declare no financial support or conflict of interest.
Data sharing statement: No additional data are available.
ARRIVE guidelines statement: The authors have read the ARRIVE guidelines, and the manuscript was prepared and revised according to the ARRIVE guidelines.
Open-Access: This article is an open-access article that was selected by an in-house editor and fully peer-reviewed by external reviewers. It is distributed in accordance with the Creative Commons Attribution NonCommercial (CC BY-NC 4.0) license, which permits others to distribute, remix, adapt, build upon this work non-commercially, and license their derivative works on different terms, provided the original work is properly cited and the use is non-commercial. See: http://creativecommons.org/licenses/by-nc/4.0/
Corresponding author: Alfredo Di Leo, MD, PhD, Full Professor, Section of Gastroenterology, Department of Emergency and Organ Transplantation, University of Bari, Piazza Giulio Cesare 11, Bari 70124, Italy. alfredo.dileo@uniba.it
Received: December 7, 2019
Peer-review started: December 7, 2019
First decision: December 30, 2019
Revised: March 6, 2020
Accepted: March 27, 2020
Article in press: March 27, 2020
Published online: April 14, 2020
Processing time: 125 Days and 20.4 Hours
Abstract
BACKGROUND

Some substances of plant origin have been reported to exert an effect in reducing intestinal neoplasm development, especially in animal models. Adenomatous polyposis coli multiple intestinal neoplasia - ApcMin/+ is the most studied murine model of genetic intestinal carcinogenesis.

AIM

To assess whether an enriched nutritional formulation (silymarin, boswellic acid and curcumin) with proven “in vitro” and “in vivo” anti-carcinogenetic properties may prevent inherited intestinal cancer in animal model.

METHODS

Forty adenomatous polyposis coli multiple intestinal neoplasia - ApcMin/+ mice were used for the study of cancer prevention. They were divided into two groups: 20 assumed standard and 20 enriched diet. At the 110th d animals were sacrificed. In each group, four subgroups received intraperitoneal bromodeoxyuridine injection at different times (24, 48, 72 and 96 h before the sacrifice) in order to assess epithelial turnover. Moreover, we evaluated the following parameters: Intestinal polypoid lesion number and size on autoptic tissue, dysplasia and neoplasia areas by histological examination of the whole small intestine, inflammation by histology and cytokine mRNA expression by real-time polymerase chain reaction, bromodeoxyuridine and TUNEL immuno-fluorescence for epithelial turnover and apoptosis, respectively. Additionally, we performed western blotting analysis for the expression of estrogen alpha and beta receptors, cyclin D1 and cleaved caspase 3 in normal and polypoid tissues.

RESULTS

Compared to standard, enriched diet reduced the total number (203 vs 416) and the mean ± SD/animal (12.6 ± 5.0 vs 26.0 ± 8.8; P < 0.001) of polypoid lesions. In enriched diet group a reduction in polyp size was observed (P < 0.001). Histological inflammation and pro-inflammatory cytokine expression were similar in both groups. Areas of low-grade dysplasia (P < 0.001) and intestinal carcinoma (IC; P < 0.001) were significantly decreased in enriched diet group. IC was observed in 100% in standard and 85% in enriched formulation assuming animals. Enriched diet showed a faster epithelial migration and an increased apoptosis in normal mucosa and low-grade dysplasia areas (P < 0.001). At western blotting, estrogen receptor beta protein was well expressed in normal mucosa of enriched and standard groups, with a more marked trend associated to the first one. Estrogen receptor alpha was similarly expressed in normal and polypoid mucosa of standard and enriched diet group. Cleaved caspase 3 showed in normal mucosa a stronger signal in enriched than in standard diet. Cyclin D1 was more expressed in standard than enriched diet group of both normal and polypoid tissue.

CONCLUSION

Our results are suggestive of a chemo-preventive synergic effect of the components (silymarin, boswellic acid and curcumin) of an enriched formulation in inherited IC. This effect may be mediated by the reduction of epithelial proliferation, the increase of apoptosis and the acceleration of villous cell renewal due to dietary formulation intake.

Keywords: Intestinal cancer; Familial adenomatous polyposis; Chemopreventive diet; ApcMin/+ mice; Boswellia; Curcumin

Core tip: A dietetic formulation containing silymarin, boswellic acid and curcumin has shown “in vitro” and “in vivo” anti-carcinogenetic properties in animal model of inflammation-related intestinal carcinoma. Herein, we assessed whether it may prevent inherited intestinal cancer in animal model (adenomatous polyposis coli multiple intestinal neoplasia - ApcMin/+). Our results showed that the dietetic formulation reduced polypoid lesion number and size on autoptic tissue, histological dysplasia and neoplasia areas. This effect is related to increased epithelial renewal and apoptosis and decreased proliferation. Our data are suggestive of a chemo-preventive synergic effect of the components of the dietetic formulation in inherited intestinal carcinoma.