الفهرس 
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Abstract
Freund’s complete adjuvant (FCA)-induced arthritis shares several characteristics with human Rheumatoid arthritis. FCA mirrors the pathology of RA, by causing hyperplasiaof the synovial tissues, inflammatory infiltration of the joints, and the destruction of bone and cartilage (Saratha and Subramanian, 2012).
Rheumatoid arthritis is characterized by synovial inflammation due to filtration of T lymphocytes, macrophages, B lymphocytes, PUFAs metabolites, and cytokines through the synovium.
Evening primrose and extra virgin olive oil exert a well known beneficial role, serving as an anti-inflammatory and antioxidant agents in several inflammatory diseases, including RA.
Forty eight rats were drawn in in the present study. These rats were separated into 8 groups (n = 6) designed as follow:
group 1: The rats within this group were received nothing except a standard balanced diet and water via ad libitum for 10 days and served as Normal control group.
group 2 (Arthritis - control 10 days post FCA injection): The rats within this group were received approximately 0.2 ml of FCA that infused into rats’ right hind footpad. No treatments applied except a standard balanced diet and water via ad libitum for 10 days.
group 3 (Arthritis - group treated with evening primrose oil for 10 days): The rats within this group were received approximately 0.2 ml of FCA that infused into rats’ right hind footpad. Evening primrose oil was given in a dose of 5 mg/kg. b. wt. by oral gavage needle with standard balanced diet and water via ad libitum for 10 days.
group 4 (Arthritis - group treated with extra virgin olive oil for 10 days: The rats within this group were received approximately 0.2 ml of FCA that infused into rats’ right hind footpad. Extra virgin olive oil was given in a dose of 5 mg/kg. b. wt. by oral gavage needle with standard balanced diet and water via ad libitum for 10 days
group 5: The rats within this group were received nothing except a standard balanced diet and water via ad libitum for 21 days and served as Normal control group.
group 6 (Arthritis - control 21 days post FCA injection): The rats within this group were received approximately 0.2 ml of FCA that infused into rats’ right hind footpad. No treatments applied except a standard balanced diet and water via ad libitum for 21 days.
group 7 (Arthritis - group treated with evening primrose oil for 21 days): The rats within this group were received approximately 0.2 ml of FCA that infused into rats’ right hind footpad. 5 mg/kg. b. wt. of evening primrose oil was given by oral gavage needle with standard balanced diet and water via ad libitum for 10 days.
group 8 (Arthritis - group treated with extra virgin olive oil for 21 days): The rats within this group were received approximately 0.2 ml of FCA that infused into rats’ right hind footpad. 5 mg/kg. b. wt. of extra virgin olive oil was given by oral gavage needle with standard balanced diet and water via ad libitum for 21 days.
At the end of each group period, rats were slaughtered under mild diethyl ether anesthesia; the ankle perimeter of the right hind legs was measured and recorded. Blood from each rat was collected from jugular vein in tubes containing ethylene-diamine-tetraacetic acid (EDTA) solution (50 ml of 15%EDTA/2.5 ml blood) for counting leukocytes and platelets, also in tubes containing noanticoagulant that left to coagulate at room temperature for 45 minutes.
Clotted blood was centrifuged at 3000 rpm for 15 minutes and the clear non-haemolysed supernatant sera were quickly removed, divided into three portions for each individual animal, and kept frozen at -20°C for consequent biochemical analysis for of hematological parameters that incude (Platelets and total leucocytic count), proinflammatory cytokine (TNF-α, IL-1, and IL-6), oxidative stress biomarkers that include (GSH content, LPO product and activity of antioxidant enzyme including GST, Gpx ), liver functions that include (ALT,and AST), and uric acid.
Spleen, thymes and hind ankle region and paw edema were immediately removed, fixed in neutral buffered formalin for 24 hrs, and then transferred into 70% alcohol for histopathological examination through the electric light microscope.
Results showed that rheumatoid arthritis caused a marked elevation in the count of platelets, white blood cells count, serum proinflammatroy cytokines, lipid peroxidation products, activities of serum enzymes of liver function (ALT, and AST), uric acid levels together with a decrease in serum antioxidants (GSH, and activity of antioxidant enzyme including GST, Gpx).
These biochemical changes in arthritic rats are confirmed by the increased ankle diameters due to the edema onset with histopathological alterations inclde focal necrosis in articular cartilage and inflammatory cells infiltration in 10 dpi and 21 dpi.
Additionally, Photomicrograph of spleen section of arthritic rats showed lymphoblasts proliferation and mitotic figure after 10 and 21 days of RA induction. Also thymus histological alterations in arthric rats showed slight lymphocytic necrosis and depletion in 10 dpi and 21 dpi groups support the biochemical changes.
The treatment with evening primrose oil for 10 and 21 days improved the deleterious effect of arthritis on cells count, inflammatory markers, oxidative stress and antioxidant defense system. This amelioration was mirrored by decreased cells count, rheumatoid factor, proinflammatory cytokines, liver enzymes, MDA product accompanied by an increase of GSH content, GST, GPx, activities.
Also, the treatment with extra vigin olive oil for 10 and 21 days enhanced the deleterious effect of arthritis on cells count, inflammatory markers, oxidative stress and antioxidant defense system. This amelioration was mirrored by decreased cells count, rheumatoid factor, proinflammatory cytokines, liver enzymes, MDA product accompanied by an increase of GSH content, GST, GPx, activities.
EPO and EVOO enhanced effects also reflected in ankle edema decrease, histological changes in joints, spleen, and thymus.
Upon comparison, for the same treatment type, the treatment for 21 days showed more obvious effect than for 10 days, extra virgin olive oil showed more explicable antiinflammtory, and antioxidant effects, than evening primrose oil.
Taken these data together, it can be concluded that rheumatoid arthritis causes inflammatory and oxidative stress which causes antioxidants depletion leading to expected hepatotoxicity. Natural products like evening primrose and extra virgin olive oil could ameliorate these deleterious effects so we recommend more studies on their combination with Disease modifying anti-rheumatic medications to improve their efficacies with less toxicity.