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石榴籽和果皮乙醇提取物的抗癌潜力及相关基因、组织学、免疫组织化学、凋亡和氧化应激特征:体外研究

Pomegranate Seeds and Peel Ethanolic Extracts Anticancer Potentials and Related Genetic, Histological, Immunohistochemical, Apoptotic and Oxidative Stress Profiles: In vitro Study.

作者信息

Nasr Mohamed, Naeem Suhaib Alsayed, El-Shenbaby Ibrahim, Mohamed Fatma Mahmoud Abdelraheem, Mahmoud Safinaz Moustafa, Abuamara Tamer M M, Abd-Elhay Wagih M, Elbayoumy Fayez Mohammed Abd Elfattah, Elkot Ahmad, Shikhon Tarek, Abo-Akrab Mostafa, Doma Mohamed A, Hasan Abdulkarim

机构信息

Histology Department, Faculty of Medicine, Al-Azhar University, Cairo, Egypt.

Clinical Pharmacology Department, Faculty of Medicine, Mansoura University, Mansoura, Egypt.

出版信息

J Exp Pharmacol. 2023 Apr 15;15:191-205. doi: 10.2147/JEP.S404321. eCollection 2023.

Abstract

INTRODUCTION

Owing to their great quantity of hydrolyzable anthocyanins and tannins, the peel and seeds of pomegranate are edible and possess potent anti-oxidant and anti-inflammatory characteristics. This work aims to trace the pomegranate seed and peel ethanolic extracts' anticancer activity against liver cancer cell line, namely HepG2 and related histopathological, immunohistochemical, genetic and oxidative stress profile.

METHODS

In vitro study for both seed and peel extract showed the prevalence of phenols, polyphenols and acids, those have anti-proliferative potential against liver cancer cell line (HepG2) with 50% inhibitory concentration (IC50) of seed significantly reduced that of peel. Toxicity of test extracts was concentration dependent and accompanied with cell cycle arrest and cell death at theG0/G1 and S phases but not at the G2/M phase. Cell arrest was supplemented with raised ROS, MDA and decreased SOD, GSH and Catalase.

RESULTS AND DISCUSSION

Apoptosis-related genes showed significant up-expression of pro-apoptotic gene (), , , and and down expression of anti-apoptotic gene (). Also, Casp-3 and P53 proteins were substantially expressed under the effect of test extracts. Histopathological study demonstrated that the untreated cells (control group) were regular cells with nuclear pleomorphism and hyperchromatic nuclei, while seed and peel extracts-treated cells showed necrosis, mixed euchromatin and heterochromatin, intra-nuclear eosinophilic structures, burst cell membranes, and the shrunken apoptotic cells with nuclear membranes and irregular cells. Finally, gene detected by immunohistochemistry was down regulated significantly under the effect of seed extract treatment than in case of cell medication with peel extract.

摘要

引言

由于石榴的果皮和种子含有大量可水解的花青素和单宁,它们可食用且具有强大的抗氧化和抗炎特性。本研究旨在追踪石榴籽和果皮乙醇提取物对肝癌细胞系(即HepG2)的抗癌活性以及相关的组织病理学、免疫组织化学、基因和氧化应激特征。

方法

对种子和果皮提取物的体外研究表明,酚类、多酚类和酸类物质含量丰富,这些物质对肝癌细胞系(HepG2)具有抗增殖潜力,种子提取物的50%抑制浓度(IC50)显著低于果皮提取物。测试提取物的毒性呈浓度依赖性,伴随着细胞周期停滞以及在G0/G1和S期的细胞死亡,但在G2/M期未出现。细胞停滞伴随着活性氧(ROS)、丙二醛(MDA)升高以及超氧化物歧化酶(SOD)、谷胱甘肽(GSH)和过氧化氢酶降低。

结果与讨论

凋亡相关基因显示促凋亡基因()、、、和显著上调,抗凋亡基因()下调。此外,在测试提取物的作用下,Casp-3和P53蛋白大量表达。组织病理学研究表明,未处理的细胞(对照组)为正常细胞,具有核多形性和核深染,而种子和果皮提取物处理的细胞显示出坏死、混合的常染色质和异染色质、核内嗜酸性结构、细胞膜破裂以及带有核膜的凋亡细胞缩小和细胞不规则。最后,免疫组织化学检测的基因在种子提取物处理的作用下比果皮提取物处理的细胞下调更为显著。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/c335/10115208/2872cddc80c5/JEP-15-191-g0001.jpg

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