الفهرس 
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Abstract
Phytochemicals are a large group of plant-derived compounds such as phenolic compounds (phenolic acids and flavonoids), terpenes, volatile oils and nitrogen compounds (indoles) hypothesized to be responsible for much of the disease protection such as cancer, cardiovascular disease,diabetes Mellitus, obesity, cataracts, aging and rheumatoid arthritis provided by diets high in fruits, vegetables, herbs, beans, cereals and plant-based beverages such as tea. Phytochemicals are referred to as chemopreventers. One of the predominant mechanisms of their protective action is due to their antioxidant activity and the capacity to scavenge free radicals. Among the most investigated chemopreventers are some vitamins, plant polyphenols, and pigments such as carotenoids, chlorophylls, flavonoids, and betalains. Resolution of the potential protective roles of specific antioxidants and other constituents of fruits, vegetables and herbs deserves major attention. Consumption of a diet high in fruits, vegetables and herbs increases antioxidant concentration in blood and body tissues and potentially protects against oxidative damage to cells and tissues. Theses antioxidants are considered as possible protection agents for reducing oxidative damage of human body from ROS and retard the progress of many chronic diseases as well as lipid peroxidation. However, more recently the polyphenols have found to be beneficial as strong antioxidants. Oxidative stress an imbalance between the production of reactive oxygen species (ROS) and antioxidant defenses, is involved in several pathological conditions such as cardiovascular or neurological disease, ENGLISH SUMMARY 122 and in aging. Reactive oxygen species (ROS), which consist of free radicals such as hydroxyl (OH-), superoxide (O2 -), nitric oxide (NO) peroxyl (RO2-), lipid peroxyl (LOO-) radicals and non-free radical species such as hydrogen peroxide (H2O2), singlet oxygen (O2-1), ozone (O3),lipid peroxide, are different forms of activated oxygen. This ROS can generate oxidative stress and produce many pathophysiological disorders
such as arthritis, diabetes, inflammation, cancer and genotoxicity. Oxidative stress and inflammation are considered significant mediators in healthy aging of the brain and in age-related neurodegenerative diseases such as Alzheimer’s and Parkinson’s disease. An in vitro study has suggested that some classes of phytochemicals also act in cell signaling and thus may protect against aging by mechanisms other than oxidative and inflammatory processes. Many academic and research centers are still paying more attention towards the toxicological effects of individual phytochemical. According to our knowledge, there is a dearth of information regarding the interactive effects of different phytochemical classes used in our daily live. Therefore, the present work was aimed to: Prepare some different extracts from plants rich with phytochemical compounds to use it in suggested formulae for aging and rheumatoid arthritis treatment. Study the toxicological effects of tested formulae through preceding many cytotoxic assays by using fish liver cells homogenate which could be used successively as in vitro biological model system in toxicological research. ENGLISH SUMMARY 123 Study the influence of the tested formulae on oxidative stress through determination the oxidant and antioxidants parameters in liver cells in vitro. To achieve these aims, the design of the study were as
follows : Mixed group of plants rich with phytochemical compounds, certain percentages and placed in capsules (each capsule 230 mg) for the protective of aging, rheumatoid arthritis and then prepare some different aqueous extracts from these plants. Prepare the fish liver cells homogenate which could be used successively as in vitro biological model system in cytotoxicity assays by NR, MTT and CV assays after that was messured oxidative stress for formulae by estimate glutathione fractions (GSH and GSSG), antioxidant enzymatic activities (GSH-Px and GSH-Rd) and biological oxidants TBARS, NO2 and NO2/NO3. The obtained results could be summarized as follow: Cytotoxicity evaluation of tested formulae by using of fish liver homogenate as biological model system. Aging formula (Herbging)The higher absorbance measurements range of NR (lysosomes activity), MTT (mitochondrial activity) and CV (cell wall membrane integrity) assays (as % of control) were (110.06-78.87, 117.43-94.62 and 104.66-96.90) for nutmeg and blackberries, while the lower one were(99.27-92.71, 96.37-84.63 and 97.44-89.75) for pomegranate and ginkgo biloba respectively. Regarding their plant parts mixture (Herbging) NR, ENGLISH SUMMARY 124 MTT and CV assays were recorded values ranged (100.57-86.91, 102.49 88.30 and 103.35-93.41) respectively. The midpoint toxicities were not recorded values (out of curve dimensions) for all the tested plant parts extracts in both individual and mixture cases. In contrary, initial point toxicities were recorded values for the all tested plant parts extract except blackberries and pomegranate at all the different assays, while ginseng and rosemary at MTT and CV assays respectively and herbging in CV assay. The highest cytotoxic value was recorded for ginkgo biloba, rhubarb and nutmeg (3.25× 10-3)while the lowest cytotoxic was ginseng (0.7× 10-1) at NR assay. On the other hand the highest cytotoxic value was ginkgo biloba (3.25× 10-3), while the lowest cytotoxic was herbging (0.3× 10-1) at MTT assay, but in CV assay the highest cytotoxic value was ginkgo biloba (3.25× 10-3) and the lowest cytotoxic were rhubarb and nutmeg (0.3× 10-1) respectively.Rheumatoid Arthritis formula (Herbrheu) The higher absorbance measurements range of NR, MTT and CV assays (as % of control) were (109.44-88.22, 117.43-94.62 and 106.00-89.91) for stinging Nettle, blackberries and their plant parts mixture(Herbrheu), while the lower one were (99.27-92.71, 96.37-84.63 and 97.44-89.75) for pomegranate and nutmeg respectively. Regarding (herbrheu) formula toxicity, it could be noticed that final toxic effect of mixture higher than the toxic effect of their component individually at CV assay. This due to the different interactive effects of herbrheu formula.
The midpoint toxicities were not recorded values (out of curve dimensions) for all the tested plant parts extracts in both individual and mixture cases. Also, the initial toxicities were not recorded values for pomegranate and blackberries at all different assays, while ginseng were ENGLISH SUMMARY 125 br>not recorded value at MTT assay. In CV assay, the initial toxicities were not recorded values except nutmeg and herbrheu formula. The highest cytotoxic value were recorded for nutmeg (3.25× 10-3, 3.25× 10-4 and 3.25× 10-3) at NR, MTT and CV assays respectively, while the lowest cytotoxic values were recorded for ginseng (0.7× 10-1) at NR assay, ginger (0.1) at MTT assay and herbrheu (0.8× 10-1) at CV assay. Amongst tested plants extracts, ginkgo biloba, rhubarb and nutmeg from formula I (herbging) and nutmeg from formula II (herbrheu) and initially induced some cytotoxic effects in fish liver homogenate. These cytotoxic effects include lysosomes and mitochondria dysfunction as well as cell wall membrane integrity, which assayed by NR, MTT, and CV assays. Changes in biological oxidative stress activity in liver cells as affected by tested formulae. Changes in glutathione fractions level in fish liver homogenate as affected by tested formulae Significant increased (p0.05) in GSH level in liver cells as a consequence of treatment with the two tested formulae. We think that different compounds found in formulae components including phenolics phenolic acids and flavonoids), Terpenoids (carnosol, carnosolic acid, rosmanol (diterpenes)), volatile oils and nitrogen compounds (indoles)etc, are antioxidants that causes an activation on the antioxidant defense potential of plasma cells. The rate of increasing of GSH level was directly proportional with the tested formulae extract concentrations. As compared with the control sample, the rate of increasing in GSH level (as% of change) were recorded 3.53, 6.44, 7.48, 17.67, 23.28 and 25.57 % ENGLISH SUMMARY 126 for herbging and 2.29, 3.74, 4.16, 9.36, 20.58 and 22.87 % for herbrheu, respectively. On the other hand the rate of decreasing in GSSG level (as% of change) were recorded 0.00, 2.38, 2.38, 2.38, 9.52 and 0.00 % for herbging, while 2.38, 0.00, 2.38, 7.14, 7.14 and 4.76 % for herbrheu for the formulae extract concentrations 10-6, 10-5, 10-4 ,10-3 ,10-2 and 10-1 , respectively. The higher ratio of increasing in reduced glutathione i.e GSH recorded in herbging than herbrheu could be interpreted by the presence of different categories of phytochemicals represent the formulae constituents. A raise in reduced glutathione (GSH) observed generally accompanied by a concomitant increased in the ratio of GSH/GSSG comparing with the control sample, the rate of increasing (as % of change) in GSH/GSSG ratio were recorded 3.56, 3.99, 5.01, 20.57, 12.59 and 25.60 % for herbging, while 0.09, 3.74, 1.74, 17.77, 12.54 and 17.28% for herbrheu formulae extract concentrations 10-6, 10-5, 10-4 ,10-3 ,10-2 and 10-1, respectively. Changes in antioxidant enzymatic components activities in fish liver homogenate as affected by tested formulae A significant increasing in GSH-Px and GSH-Rd activities was observed in liver cells as a consequence of treatment with the tested formulae. The rate of increasing of antioxidant enzymes was directly proportional with increasing of tested formulae extract concentrations. As compared with the control sample, the rate of increasing (as % of change) were recorded in GSH-Px 15.37, 18.16, 20.56, 40.32, 36.13 and 43.51% for herbging formula and 3.39, 5.59, 14.57, 19.36, 19.96 and 54.89 for herbrheu formula, respectively. In GSH-Rd enzyme the rate of increasing (as % of change) were recorded 1.63, 2.28, 20.85, 35.50, 32.90 and ENGLISH SUMMARY 127 37.79 % for herbging formula and 2.93, 9.77, 12.05, 15.96, 26.06 and
33.88 for herbrheu formula, respectively. The higher ratio of increasing in antioxidant enzymes recorded in herbging and herbrheu formulae could be interpreted by the presence of different categories of phytochemicals represent the formulae constituents Changes in biological oxidants levels in fish liver homogenate as affected by tested formulae Significant decreased (p 0.05) in TBARS, NO2 and NO2/NO3 levels in liver cells as a consequence of treatment with the tested formulae. The rate of decreasing of biological oxidants was directly proportional with increasing of tested formulae extract concentrations. As compared with the control sample, the rate of decreasing (as % of change) was recorded in TBARS (2.07, 3.05, 8.68, 13.87, 16.77 and 21.56 %), while in NO2 the rate of decreasing (as % of change) were recorded (5.33,8.78, 16.93, 17.87, 19.12 and 24.45 %) and in NO2/NO3 (0.17 1.51 7.29, 22.63, 32.86 and 28.50 %) for herbging formula, respectively. In herbrheu formula the rate of decreasing (as % of change) were recorded in TBARS (