الفهرس 
IntroductionOnly 14 pages are availabe for public view
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Abstract
Salvadoraceae Is A Plant Family Composed Of Three Genera And With A Total Of Eleven Known Species. The Species Belonging To Genus Salvadora Are Trees Or Shrubs With Opposite Leaves, Flowers Minute, 4-Merous In A Rich Terminal Panicle. Fruit A Small, White Or Pale Purplish, Globose Drupe Of Pungent Taste Like The Leaves.
Salvadora Persica L. Is A Much Branched Shrub Or Small Tree, Glabrous In All Its Parts, Covering Sometimes An Area Of 27 M. Leaves Are Oblong, 5-6 Cm Long.
This Study Inlcudes Two Parts:
Part I: Phytochemical Investigation Including:
• Chapter (1) Preliminary Phytochemical Screening. The Preliminary Phytochemical Screening Of The Total Ethanolic Extract Of The Arieal Parts Of Salvadora Persica L. Showed The Following:
o Presence Of Carbohydrates Or Glycosides, Alkaloids And /Or Nitrogenous Compounds, Saponins, Sterols And /Or Triterpenes, Tannins And Volatile Oils.
o Absence Of Sublimable Compounds, Oxidase Enzyme, Cardiac Glycosides And Cyanogenetic Glycoside.
• Chapter (2) Extraction, Successive Fractionation Of The Plant Extract & Metabolomic Profiling Of The Ethanolic And The Aqueous Fraction Of The Arieal Parts.
The Weight Of Residue Left After The Extraction Of Aerial Parts Of S. Persica (Leaves And Stems) Were Extracted Using 95% Ethyl Alcohol, Then The Dried Ethanolic Extracts (297 Gm) Were Further Suspended In The Least Amount Of Distilled H2O Transferred To Separating Funnels And Fractionated Separately Using Different Organic Solvents (Pet. Ether, Dichloromethane And Ethyl Acetate), Then The Remaining Mother Liquors Were Concentrated To Afford Four Successive Fractions from The Plant [Pet. Ether (39mg), DCM (20mg), Ethyl Acetate (10 Mg) And Aqueous (228 Mg)] Fractions. The Crude Ethanolic Extract And Their Derived Fractions Of The Plant Under Study Were Then Subjected To LC/MS Analysis. Ethanolic And Aqueous Extracts Of S. Persica Were selected For Metabolomic Analysis And Different Biological Activites.
Metabolomic Investigations Of The Ethanolic And The Aqueous Extracts Of The Aerial Parts Of Salvadora Persica L.
The Crude Extract As Well As The Aqueous Fraction Of The Aerial Parts Of S. Persica Were Subjected To Metabolomic Profiling, For The First Time, Using LC-HR-ESI-MS For Dereplication Purposes. The Detected 28 Compounds Were Tentatively Identified By Employing Macros And Algorithms That Coupled Mzmine With Online And In-House Databases, I.E., METLIN And DNP Databases For Secondary Metabolites Of Higher Plants.
Metabolic Analysis Also Revealed The Presence Of 28 Compounds And 11 Compounds Of The Total Ethanolic And The Aqueous Extract; Respectively Such As Naringenin And Volatile Constituents Such As Eugenol, Carvacrol, Thymol, Β-Pinene And Eucalyptol. Seven Nitrogenous Compounds Were Identified As Benzylamine,Aniline,Benzylurea,Butanediamide,Benzylisothiocyanate,Nbenzyl-2- Phenylacetamide, And Theobromine.
• Chapter (3) Quantitative Determination Of Total Flavonoid Contents Of The Aqueous Fraction.
O Total Flavonoid Content In The Aerial Part Of S. Persica L. Was 6.2 Mg Rutin Equivalent.
Part II: Biological Study Including:
• Evaluation Of Antioxidant Activity Of The Aqueous Fraction.
The Total Antioxidant Capacity Of The Petroleum Ether, Dichloromethane, Ethylacetate And Aqueous Fractions Of S. Persica Were Determined. The Results Were (1.22%, 7.86%, 8.96% And 37.8%) Scavenging Capacity Respectively. Results Revealed That Aqueous Fraction Exhibited The Highest Antioxidant Capacity With IC50 = 24.54±0.9 µg/Ml. This Result Can Explain That The Highest Percent Scavenging Capacity Was Produced By The Aqueous Fraction.
• Molecular Docking Study Of The Flavonoidal Compound Into The Active Site Of Different Active Sites To Illustrate The Binding Energy Of The Compounds And Affinity To Different Ligands And Different Biological In Vitro Study As Follows:
Results In That Generally, Most Of The Target Compounds And The Ligand Showed Comparable Binding Pattern And Equal Docking Score Can Be Used To Compare The Binding Affinity Of Different Ligands To The Same Protein.
Concerning To The Anticholinesterase Study, The Molecular Docking Study Of Identified Flavonoidal Compounds Into All Compounds Identified In The Aqueous Extract Showed High Affinity Into The Active Site Of Acetylcholinesterase Enzyme where Compounds 8, 5,1 Showed The Highest Activity.
In Vitro Study Of The Aqueous Extract And Its Silver Nano Formulation As Anticholinesterase Activity.
On Rabbit Ileum
The Aqueous Extract Of S. Persica Increased The Action (Contraction) Of Acetylcholine On The Rabbit Ileum By Dose Of Acetylcholine from 5µg To 100µg And The Effect Was Dose Dependent
Spectrophotometric Assay
Total Ethanolic Extract Of S. Persica Showed 55% Inhibition Of Acetyl Cholinesterase Enzyme Activity When Compared To Control, While Petroleum Ether, Dichloromethane, Ethyl Acetate And Aqueous Fractions Showed 9%, 8%, 6% And 20% Inhibition, Respectively.
As Aqueous Fraction Showed The Highest Percent Of Inhibition (20%) With IC50 = 79.4±1.6 µg/Ml, Silver Nitrate Nanoparticles Were Prepared from This Fraction. Nanomolecules Gave 56% Inhibition With IC50=10.71±2.1 µg/Ml Which Was Higher Than That Of The Inhibition Produced By The Aqueous Fraction.
Concerning To The Molecular Docking Study Of Identified Flavonoidal Compounds Into Main Protease Active Site Of COVID 19 Virus Using Darunavir® As A Ref. Drug All Identified Flavonoid Compounds Except 9 Were Better Than The Currently Used COVID-19 Main Protease Inhibitor Darunavir And Flavonoids 5, 7, 3 And 4 In order Showed The Highest Stability In The Active N3 Binding Site.
Concerning To The Molecular Docking Study Of Identified Flavonoidal Compounds Into The Contact Surface Of Hace2 Corona Virus - Spike Protein Complex Most Compounds Of The FRF Showed Prominent Interaction With Both Hace2 And The Viral Spike Protein And The Most Active Compound 2 Was Mauritiannin Comparing To Hispiridin As A Reference Drug Or Positive Control Inhibitor.
In Vitro Study Of The FRF And Its Liposomal Formulation Against SARS-COV-2
In Vitro RT-PCR Testing
FRF And Its Liposomal Formulation Led To A High Significant Increase In The Percentage Inhibition Of Viral Replication from 38.09 ± 0.83% In FRF To 85.56 ± 1.12% In FRF-Lip Comparing To The Ref. Drug Remdesivir® Which Has (91.20±1.71%) Percentage Inhibition Of Viral Replication. This Could Be Attributed To The Higher Surface Area And Better Dispersibility Of The Liposomal Formulation Which Was Able To Prevent Precipitation And Aggregation Of The Lipophilic Components. Hence, It Could Optimize The Expected Poor Cellular Uptake Of The Hydrophilic Components, And Enhance The Delivery Of Such Challenging Natural Products.
Concerning To The Molecular Docking Study Of Identified Flavonoidal Compounds Into The The Active Site Of NS3/4A Protease Of Hepatitis C Virus (HCV) All Compounds Have Comparable Affinity To The selected Pocket According To Binding Affinity. Notably Compounds 10, 6, 1 And 8 In order Showed Good Binding Affinity Energies If Compared To Co-Docked Ligand Vaniprevir As Positive Control Inhibitor.
Concerning To The Molecular Docking Study Of Identified Flavonoidal Compounds Into The The Active Site Of NS5B Polymerase Of Hepatitis C Virus (HCV) All Compounds Have Good Affinity To The selected Pocket According To Binding Affinity. Notably Compounds 1, 4, 5 And 11 In order Showed Good Binding Affinity Energies If Compared To Co-Docked Ligand Sofosbuvir As Positive Control Inhibitor.
Concerning To The Molecular Docking Study Of The Identified Flavonoidal Compounds Into The The Active Site Of HIV-1 Protease Virus All Compounds Have Comparable Affinity To The selected Pocket According To Binding Affinity. Notably Compounds 5, 6, 7 And 2 In order Showed Good Binding Affinity Energies If Compared To Co-Docked Ligand Tipranavir As Positive Control Inhibitors.
In Vitro Study Of The Aqueous Extract And Its Liposomal Formulation Against Hepatitis C Virus (HCV) And Human Immune Deficiency Virus (HIV)
The Prospective Anti-HIV And Anti-HCV Effect Of The Aqueous And Its Liposomal Formulation Was Investigated By In Vitro RT-PCR Testing Which Specially Detects HIV And HCV RNA Quantitatively.
Concerning To HIV Liposomal Formulation Of The Aqueous Extract
Showed A High Significant Increase In The Percentage Of Inhibition In Viral Replication from 42.81% To 87.95% Comparing To The Reference Drug Vaniprevir® Which Has A Percentage Of Inhibition 96.24 %.
Concerning To HCV Liposomal Formulation Of The Aqueous Extract
Showed A High Significant Increase In The Percentage Of Inhibition In Viral Replication from 32.42% To 84.03% Comparing To The Reference Drug Sofosubvir® Which Has A Percentage Of Inhibition 93.44 %. It Was Useful To Find A Natural Product Having The Ability To Inhibit HIV And HCV Replication In A Value Very Close To That Of The FDA-Approved Anti-HIV And Anti-HCV: Sofosbuvir® And Ribavirin®.
Concerning To The Cytotoxicity Study Against Melanoma Cell Line Of The Aqueous Extract With (IC5024.5±9) In Comparison To The Reference Drug Doxetaxel With (IC507.73±6) Which Indicates That The Aqueous Extract Has A Cytotoxic Effect On The Cell Line As Anticancer Drug But Its Potency Is About 31.5 % Of The Ref. Standard Drug.
Concerning To In Vivo Study Of The Cardioprotective Effect Of The Ethanolic Extract.
Serum Biomarkers Indicates That The Total Ethanolic Extract Significally Decreases The Levels Of Lactate Dehydrogenase (LDH), Creatine Kinase Iso‐ Enzyme MB (CK-MB) And Troponin I Leading To Decreasing The Cardiotoxic Effect.
Immunohistochemical Markers Illustrate That The Most Cardiac Muscle Fibers Retained Their Acidophilic Sarcoplasm, Little Inter Fiber Space And Less Dilated Blood Vessels Indicating The Cardioprotective Effect Of Tatal Ethanolic Extract.
Immunohistochemically Stained Cardiac Tissue Reveals That:
Concerning BCL2 (Marker For Antiapoptitic) group IV (Receiving Doxorubicin + Extract) Showing Apparent More Cytoplasmic Immunoreactivity In More Cardiomyocytes Comparing To group III (That Received Doxorubicin Alone) Showing Less Cytoplasmic Immunoreactivity In Few Cardiomyocytes.
Concerning LC3 (Marker For Injured Cells Or Phagosomes Activator) group III Showed A Significance Increase In The Mean Number Of Immunostained LC3 If Copmared To Groupi And II. While group IV Showed A Significant Decrease In This Value If Compared To group III.