الفهرس 
INTRODUCTIONOnly 14 pages are availabe for public view
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Abstract
The recent research focuses on the use of natural products derived
from plants in offering protection against the harmful effects of radiation.
Reactive oxygen species (ROS) are the cause of certain diseases, such as
cancer tumors, cardiovascular diseases, and liver dysfunction. Ionizing
radiation passing through living tissues generates free radicals that can
induce DNA damage. The damaging effects of ionizing radiation on DNA
lead to cell death and are associated with an increased risk of cancer
Ionizing radiation finds applications in many fields of the medical
sciences and enhances the efficacy of diagnostic and therapeutic uses.
Radiotherapy has been used as a treatment modality for cancer patients
during their course of illness. Irradiation has associated side effects as it
induces damage to normal tissue, resulting in acute and chronic toxicities
or severe organ dysfunction. Hence, the need arises to protect human
beings from the effects of radiation, even in a planned or unplanned
accidental exposure.
Even though the clinical application of ionizing radiation in
radiotherapy and other medical areas is widely accepted and has accrued
colossal success, the damage to the healthy surrounding tissues has limited
its usage. Thus, there is an urgent need to develop drugs from plants and
herbs that are cost-effective, readily available, and capable of scavenging
free radicals, thus protecting normal cells during radiotherapy and reducing
radiation’s harmful effect in an emergency radiation accident. This is the
motivation for this work.
Sargassum algae are potential sources of antioxidant agents such as
polyphenols, including flavonoids and phenols, amino acids, fatty acids,
and essential elements as well as other classes of secondary metabolites
such as protein, proanthocyanidins, beta-carotene, and saponins, which are
of high nutritional value and important to humans and animals.
In this study we prepared nano metal complex of copper acetate with
ahydrazide ligand and nano metal oxides of(ZnO, Fe2O3, and CuO )at a
concentration of 10 μM with seaweed Sargassum vulgare nano methanolic
extract at a concentration of 4 g/kg body weight of the rat. Also, we studied
the characterization of these nanoparticles by using different techniques
like X-Ray diffraction analysis (XRD), scanning electron microscope
(SEM), fourier transform infrared spectroscopy (FTIR), diffuse reflectance
spectra (DRS), and energy dispersive X-ray (EDX).
The present study has been carried out in order to access the radioprotective efficiency role and the antioxidant effect of natural biochemical
ingredients present in different groups of rat treated with Sargassum
vulgare extract loaded on nano metal particles (ZnO, Fe2O3, [Cu(OAC)2
(HL)2]complex and CuO) against oxidative stress and liver tissue injury in
the gamma-irradiation (6 Gy)-treated group of rats.
Algal sampling, identification and extraction
Specimens of the brown seaweed Sargassum vulgare (Fucales,
Phaeophyta) were collected on October 2021 from the littoral zone of rocky
shorelines in Hurghada city (27° 15′ 58.45″ N, 33° 48′ 57.09″ E), the Red
Sea coast of Egypt. it is one of the most important places of interest for
algal growth in Egypt (intertidal zone).
Metabolomic profiling and antioxidant properties of Sargassum
vulgare
Polyphenol fractions (flavonoids and phenolics), phytochemical
constituents (proanthocyanidins, saponins, β-carotene, and protein),
antioxidant capacity ABTS˙⁺[2,2-azino-bis (3-ethylbenzothiazoline-6-
sulfonic acid)], [DPPH˙(2,2-diphenyl-1-picrylhydrazyl), radical scavenging
activity, and ferric-reducing antioxidant power (FRAP)], fatty acids, amino
acids, mineral and elemental contents in seaweed Sargassum vulgare, were
investigated.
Experimental design and animal treatment
One hundred and twenty adult males (13 weeks old), Wistar albino
male rats (n=15X8) weighing about 180-200 g were brought from Research
Institute of Ophthalmology (animal house), Egypt, housed in cages
(stainless steel) with good ventilated shields, the rats were acclimated with
standard experiment conditions (26±2°C, 12/12 h light/dark, and 65% RH)
for one week before the start of the experiment. During the whole
experiment, all the animals were fed on well-balanced diet (3.5% fats, 22%
proteins, 0.55 table salt, 0.72% molasses, 0.25% vitamins, 60% corn maize,
5% wheat bran, 20% soybeans, and 10% growth additives) and free access
to drinking water add libitum for six weeks.
Rats were randomly split into eight groups:
The rats were randomly divided into eight groups (15 rats each).
group I served as normal control rats that were received normal food
and water for six consecutive weeks daily.
group II Rats subjected to a single dose (6 Gy) of whole body
gamma irradiation after one week of the experiment.
group III Rats were treated intraperitoneally with seaweed
Sargassum vulgare nano methanolic extract at a concentration of 4 g/kg
body weight.
group IV Rats subjected to a single dose of γ-irradiation (6 Gy) after
one week of the experiment, then injected intraperitoneally with seaweed
Sargassum vulgare nano methanolic extract at a concentration of 4 g/kg
b.wt. twice/week for six consecutive weeks.
group V Rats subjected to a single dose of γ-irradiation (6 Gy) after
one week of the experiment, then injected intraperitoneally with seaweed
Sargassum vulgare methanolic extract with nano ZnO twice/week for six
consecutive weeks.
group VI Rats subjected to a single dose of γ-irradiation (6 Gy) after
one week of the experiment, then injected intraperitoneally with seaweed
Sargassum vulgare methanolic extract with nano Fe2O3 twice/week for six
consecutive weeks.
group VII Rats subjected to a single dose of γ-irradiation (6 Gy) after
one week of the experiment, then injected intraperitoneally with seaweed
Sargassum vulgare methanolic extract with nano CuO twice/week for six
consecutive weeks.
group VIII Rats subjected to a single dose of γ-irradiation (6 Gy)
after one week of the experiment, then injected intraperitoneally with
seaweed Sargassum vulgare methanolic extract with nano [Cu(OAC)2
(HL)2]complex twice/week for six consecutive weeks.
Blood and tissue preparation samples
Six weeks after the experiment was completed, rats were lightly
anesthetized and blood was collected from the internal jugular vein in
special tubes. Separated clear sera were kept at -20°C for further
biochemical analysis. Then, under deep anaesthesia all rats were sacrificed.
Immediately, carefully remove the liver from each rat for all eight groups
for histopathological examination.
The parameters studied in the current work were inflammatory
biomarkers [tumor necrosis factor-α (TNF-α), total heat shock protein
(HSP70), interleukin-6 (IL-6), interleukin-10 (IL-10), caspase-3 (CASP-3),
monocytes chemoattractant protein-1 (MCP-1), matrix metalloproteinase-9
(MMP-9), and 8-hydroxy-deoxyguanosine (8-OHDG)], contents of nitric
oxide (NO), deoxyribonucleic acid (DNA), malondialdehyde (MDA),
antioxidant enzymes (glutathione (GSH), catalase (CAT), superoxide dismutase
(SOD), enzyme activity of alanine aminotransferase (ALT), aspartate
aminotransferase (AST), alkaline phosphatase (ALP), alpha feto protein (AFP),
and gamma-glutamyl transferase (GGT), total bilirubin, albumin, total
protein, globulin, cholesterol, triglyceride, hematological parameters
(haemoglobin (HB), red blood cells (RBCs), white blood cells (WBCs),
platelet), kidney functions (creatinine, urea, potassium and sodium), hormonal
level of follicle-stimulating hormone (FSH), luteinizing hormone (LH), and
testosterone in different rat groups were determined.
The results of the present study were summarized as follows:
1- Rats subjected to γ-irradiation (6 Gy)+Sargassum vulgare with
nano [Cu(OAC)2 (HL)2]complex (GVIII) was the most effective treatment
in reducing levels of ALT, AST, and AFP as compared to the other groups.
Meanwhile, Rats subjected to γ-irradiation (6 Gy)+Sargassum vulgare with
nano CuO was the most effective treatment in reducing levels of total
bilirubin and AFP (GVII).
Also, they increased the level of total protein, albumin, and globulin
as compared to the gamma irradiation groups. This is attributed to the
increased synergistic activity of the powerful antioxidant polyphenolic
compounds (radical scavenging properties) as well as other additional
secondary metabolites.
2- The gamma irradiation treatment (6 Gy) increased the concentrations
of creatinine, urea, and potassium as compared to the control group. In contrast,
gamma irradiation (6 Gy) significantly decreased sodium levels when
compared to all treatment. However, Rats subjected to γ-irradiation (6
Gy)+Sargassum vulgare with nano CuO (GVII) was the most effective
treatment in reducing levels of creatinine, Urea, and potassium as compared to
the γ-irradiated group. Also, they increased the level of sodium as compared
to the γ-irradiated group. This is attributed to the increased synergistic
activity of the powerful antioxidant polyphenolic compounds (radical
scavenging properties) as well as other additional secondary metabolites
3- Gamma irradiation treatment (6 Gy) increased cholesterol, and
triglycerides as compared to the control group. Meanwhile, application of γirradiation (6 Gy)+Sargassum vulgare with nano CuO (GVII) was the most
effective treatment in decreasing cholesterol, and triglycerides levels as
compared to γ-irradiated group.
4- The gamma irradiation treatment (6 Gy) decreased the concentrations
of SOD, GSH, and CAT as compared to the control group. In contrast, gamma
irradiation (6 Gy) significantly increased MDA and NO levels when
compared to all treatment. However, Rats subjected to γ-irradiation (6
Gy)+Sargassum vulgare with nano CuO (GVII) was the most effective
treatment in increasing the levels of SOD, GSH, and CAT as compared to the
γ-irradiated group. Also, they decreased the level of MDA and NO as
compared to the γ-irradiated group.
5- Gamma irradiation treatments increased significantly all the
inflammatory biomarkers as compared to the control group and Sargassum
vulgare group. However, Rats subjected to γ-irradiation (6 Gy)+Sargassum
vulgare with nano CuO (GVII) was the most effective treatment in
decreasing significantly TNF-α, HsP70, CASP-3, MMP-9, IL-6, IL-10, and
MPC-1, as compared to the gamma irradiated group.
6- Gamma irradiation treatment (6 Gy) decreased HB, RBCs, WBCs,
and platelets as compared to the control group. The results indicate that the
seaweed Sargassum vulgare is nontoxic to the hematopoietic system. In the
present study, we assessed the application of S. vulgare methanolic extract
(4 g/kg b.wt.) loaded on nano metal particles (ZnO, Fe2O3, [Cu(OAC)2
(HL)2]complex and CuO) ability to mitigate radiation-induced
hematological disorders in rats. Rats subjected to γ-irradiation (6
Gy)+Sargassum vulgare with nano CuO (GVII) was the most effective
treatment in increasing the HB, WBCs, and platelets as compared to the
gamma irradiation treatment.
7- There are many degenerative effects in the liver tissues in gamma
irradiated group (6 Gy). Histopathological examination showed to some
extent normal structures of the liver in both normal control rats, group of
methanolic extract of sargassum only, and gamma irradiated group (6 Gy)
treated with S. vulgare methanolic extract (4 g/kg b.wt.) loaded on nano
metal particles (ZnO, Fe2O3, [Cu(OAC)2 (HL)2]complex and CuO), and
decreased significantly most of the liver tissue injury.
CONCLUSION
The results obtained from the research study concluded that
Sargassum vulgare can be used as a promising functional food ingredient
to protect patients and workers during radiation therapy and as a natural
therapeutic drug due to its valuable nutritional benefits, safe nature, low
cost and also the availability and availability of the moss.
Deficiency of dietary supplementation in patients can be overcome
with Sargassum vulgare, which may have significant health-promoting
potential (potential protective agent) against gamma irradiation-induced
toxicity in rats.
Associated with the synergistic activity of potent antioxidants, such as
polyphenols (free radical scavenging properties), B-complex vitamins,
fiber, and essential elements like Fe, Mn, Co, Se, Cu, and Zn which are
known to be very important for various metabolic processes in the human
body as they are known to be closely linked to growth and health of human
being, carbohydrates, fatty and amino acids, proanthocyanidins, saponins,
β-carotene, and protein which are important for a good healthy diet to
alleviate and ameliorate different apoptotic and inflammatory oxidative
stress patterns associated with exposure to gamma radiation.
The great health-promoting potential of the macroalga S. vulgare
against gamma radiation associated toxicity seems to be due to the
synergistic activity of its potent antioxidant phenolics (mainly gallic,
ferulic, salicylic, and chlorogenic and p-coumaric acids) and flavonoids
(rutin, kaempferol, and hesperidin), as well as other additional polar
compounds. These natural antioxidant compounds are distinctly capable of
scavenging ROS and subsequently improving the cellular
oxidant/antioxidant balance, modulating the immune defense systems, and
eventually regulating the signaling pathways that lead to initiation of the
proinflammatory cytokines and apoptotic proteins.
The preliminary phytochemical analysis for Sargassum vulgare
revealed the presence of many active constituents that may be
pharmacologically beneficial. The animals were found to be free of any
major toxic signs during as well as at the end of the acute and subchronic
toxicity study. There were no abnormal signs of any motor or sensory
functions.
Natural antioxidant compounds present in Sargassum vulgare are
clearly capable of scavenging ROS and thereafter strongly improving the
cellular balance (oxidant/antioxidant process), preserving normal liver
functions, decreasing lipoperoxiation of cells, improving the immune
defence systems, increasing antioxidant enzyme activities, and finally
downregulating the singalling pathways responsible for the initiation of
apoptotic proteins and proinflammatory cytokines. The application of the
methanolic extract of Sargassum vulgare with nano-copper oxide, ferric
oxide, zinc oxide and copper(II) acetate complex in the presence of the
group treated with gamma rays (6 Gy) led to a significant decrease in most
liver tissue injuries, which was somewhat similar to the control group.