Phytochemical and molecular characterization of active constituents of Lycium barbarum and their effects on normal human lymphocytes and cancer cell line
Other Title(s)
الخصائص الكيميائية و الجزيئية للمكونات الفعالة لنبات العوسج Lycium barbarum و تأثيرها في الخلايا اللمفاوية البشرية الطبيعية و الخلايا السرطانية
Dissertant
Thesis advisor
al-Samirrai, Khulud Wahib Abbud
Hamzah, Subhi Jawad
Comitee Members
Umran, Mahfuzah Abbas
Shukr, Sad M.
Jasim, Abd al-Jasim M.
Adhiah, Ali Husayn
University
University of Baghdad
Faculty
College of Science
Department
Biotechnology Department
University Country
Iraq
Degree
Ph.D.
Degree Date
2013
English Abstract
The discovery and identification of a new drugs, which can potentiate the immune function has become an important goal of researches in immune-pharmacology and oncotherapy.
This study demonstrates the favorable effects of Iraqi wild type Lycium barbarum active components as anticancer and immunomodulating agent.
Flavonoids from Lycium barbarum leaves were extracted and identified by two chromatographic methods; the preparative thin layer chromatography (PTLC), and column chromatography.
The PTLC technique was the best toolto separate several flavonoids represented in; Rutin, Quercetin, Kaempferol, luteolin and a major quantity of unknown flavonoid that had been separated and purified to evaluate its biological activity.
Iraqi L.
barbarum leaves contain total flavonoids (11.28 mg/g dried leaves) calculated as Quercetin.
The fruit of Iraqi Lycium barbarum is rich with Polysaccharides and Carotenes, both of which were investigated qualitatively and quantitatively in the present study.
The extracted polysaccharides total content calculated as glucose was 3.4mg/g dried fruits.
The average plant content of total carotene was 0.31mg/g dried fruit as determined by two methods: high performance liquid chromatography (HPLC) and Standard β-carotene curve.
The three extracted components from Lycium barbarum (purified flavonoid, polysaccharides, and carotenes) showed cytotoxic effect towards both: the primary cell culture of normal hepatic cells (WRL-68), and cancer hepatic cell lines (HepG-2) at 100μg/ml concentration for 24 hours treatment.
Purified flavonoid exerted the potent effect on both cell lines among the other two extracted components.
The High Content Screening (HCS) assay was held only for the purified flavonoid to investigate the ii mechanism by which the purified flavonoid affected living cells toward apoptosis.
The most significant reduction (p≤0.05) in cell viable count was at the concentration 100μg/ml which appear to cause the induction of cell death via mitochondrial pathway for HepG-2 cell line after 24 hours exposure.
The purified flavonoid had no effect on the HepG-2 cell cycle.
The Immunomodulation effect for the purified flavonoid and the extracted polysaccharides on normal human peripheral blood lymphocytes showed that the purified flavonoid suppress lymphocytes proliferation, while the later increased lymphocytes proliferation significantly.
The immune stimulating effect of the polysaccharides caused alteration in IL-10 and TNF-α level estimated by ELISA technique towards IL-10 elevation for the normal human blood lymphocytes against TNF-α level after 2 hours exposure at concentrations (250 and 500 μg/ml), while apposite results were shown after 4 hours of exposure.
The percentage of CD4+ and CD8+ markers cells was calculated for the two interval times of the lymphocytes treated with different polysaccharides concentrations.
The CD8+ cells percentage increased at 250μg/ml concentrations after 2 hours treatment, while after 4 hour exposure CD4+cells percentage elevated at 500μg/ml concentrations.
Polysaccharides effects on IL-10 and TNF-α production indicated by alteration in gene expression of normal lymphocytes in vitro toward IL-10 production in response to all concentrations(125, 250 500)μg/ml for 2 and 4 hours intervals, while the extract polysaccharides showed no effect on TNF-α gene expression for both time intervals.
Main Subjects
Topics
No. of Pages
144
Table of Contents
Table of contents.
Abstract.
Abstract in Arabic.
Introduction.
Chapter One : Literature review.
Chapter Two : Materials and methods.
Chapter Three : Results and discussions.
Conclusions and recommendations.
References.
American Psychological Association (APA)
Muhammad, Zaynab Yasin. (2013). Phytochemical and molecular characterization of active constituents of Lycium barbarum and their effects on normal human lymphocytes and cancer cell line. (Doctoral dissertations Theses and Dissertations Master). University of Baghdad, Iraq
https://search.emarefa.net/detail/BIM-601434
Modern Language Association (MLA)
Muhammad, Zaynab Yasin. Phytochemical and molecular characterization of active constituents of Lycium barbarum and their effects on normal human lymphocytes and cancer cell line. (Doctoral dissertations Theses and Dissertations Master). University of Baghdad. (2013).
https://search.emarefa.net/detail/BIM-601434
American Medical Association (AMA)
Muhammad, Zaynab Yasin. (2013). Phytochemical and molecular characterization of active constituents of Lycium barbarum and their effects on normal human lymphocytes and cancer cell line. (Doctoral dissertations Theses and Dissertations Master). University of Baghdad, Iraq
https://search.emarefa.net/detail/BIM-601434
Language
English
Data Type
Arab Theses
Record ID
BIM-601434
