THE EFFECT OF GINGER TINCTURE AND OLIVE OIL ON THE EXPERIMENTALLY INDUCED BONE MARROW DEPRESSION IN ADULT MALE ALBINO RAT

Background: Cyclophosphamide (CP) is a nitrogen alkylating agent used in various types of cancer chemotherapy, in spite of that CP chemotherapy induces sever bone marrow damage and depression. Many natural compounds derived from plants proved to have a protective effect against the toxic effects of many chemicals, such as ginger plant which is considered as a rich source of antioxidants, while olive oil plays a well established, beneficial role in promoting good health. Material and method: Forty adult male albino rats were divided into four equal groups: GI; control group, GII; ginger and olive oil receiving group, GIII; CP injected group, GIV; group receiving both CP with ginger and olive oil. This experiment continued for 15 days. At its end blood samples were collected for complete blood picture and bone marrow samples were taken for histological, immunohistochemical and chromosomal studies. Morphometric and statistical studies were also done. Results: CP caused prominent deficiency in the hemopoietic cells within the bone marrow, it decreased the RBCs, WBCs and platelets count in peripheral blood. CP produced depression of dividing bone marrow cells. This was detected by PCNA immunohistochemical study. Also, it produced some chromosomal structural changes. All these changes were improved when CP given in association with ginger tincture and olive oil. Conclusion: Administration of ginger tincture and olive oil is playing a role in protection of the body against the harmful effects of some toxic agent as CP. KEY WORD: cyclphosphamide, ginger, olive oil, bone marrow and chromosomes. Selim et al. European Journal of Biomedical and Pharmaceutical Sciences www.ejbps. com 2 properties. It could release chronic diseases such as cardiovascular disease, atherosclerosis, and some types of cancers – especially colorectal and breast cancer. [8] Therefore, the aim of this work is to study the histological, immunohistochemical and cytogentic changes of bone marrow after exposure to CP and the possible protective effect of both ginger tincture and olive oil in adult male albino rat. MATERIAL AND METHOD Forty adult male albino rats, weighing 200g±10g, were used in this experiment. They were housed in ventilated cages in the animal house of histology department, Faculty of medicine for Girls, Al-Azhar University, Cairo, Egypt. Animals were given food (normal rat chow) and free water access. Animals were kept for one week before the beginning of the experiment for acclimatization. The animals were randomly divided into four equal groups; N=10. Group I: control group. Group II: ginger and olive oil group, in this group the animals received daily oral dose of both ginger tincture and olive oil. Olive oil and fresh ginger were purchased from market. They were given to the animals by gastric tube. The dose of olive oil was 1ml/100 gm body weight [9] while the ginger tincture was 250 mg/kg body weight [7] . The ginger tincture was prepared daily for each chosen rat by adding 50 mg of fresh minced ginger to 2 ml cold water, this mixture was prepared at afternoon, left in refrigerator till the morning of the next day, where it is filtered and given to the animal. Group III: CP group, the animals received CP by intraperitoneal route of administration. The dose of CP was 40mg/Kg body weight alternatively for 15 days. CP was dissolved in distilled water (200mg/ 20ml) [10] . The CP was purchased from market. Group IV: CP and ginger olive oil group, in this group the animals were given both CP, ginger tincture and olive oil by the previously prescribed doses and route of administration. The experiment was continued for two weeks, after this duration half the number of rats from each group was anesthetized by ether inhalation. Then, the blood samples were collected from retro-orbital venous plexus for complete blood picture which was done in Al-Azhar University Center for Virsus Research and Studies. Bone marrow of both femora from each rat were collected and fixed in Bouin’s solution [11, 12] . Then, processed for paraffin blocks formation, serial sections of 5μ thickness were cut and stained with H&E for routine histological examination of bone marrow structure [13] . The slides were examined under the light microscope and the images were taken with microscope connected to digital camera. For immunohistochemical study, avidinbiotin peroxidase technique for proliferating cell nuclear antigen (PCNA) [14] using PCNA antibodies (DakoDenmark, diluted 1:100). PCNA antibodies served as a reliable marker of proliferation [15] . Cytogenetic study was done on the remaining half of rats from each group. This half of rats was injected colchicine via intraperitoneal route 2 hours prior to sacrifice. The colchicine is an antimitotic drug. Bone marrow from all groups of animals were collected in freshly prepared hypotonic solution of potassium chloride (KCl), the e suspension w s in u ted for in t then entrifuged the supern t nt w s discarded, then the cells were resuspended in cold fixative of methanol/acetic acid (3:1). The centrifugation and fixation were repeated twice at an interval of 30 min., finally the cells were resuspended in fixative and were spread by dropping on slides. The slides were flame dried and stained with 10% buffered Giemsa stain. [16] Morphometric and statistical studies were done for the following: i) The mean number of PCNA positive cells/ high power field was measured in five fields per slide. Five slides were used for each group [17] . ii) The number of chromosomal aberrations was counted in 50 metaphase spread in each group under 1000x (oil immersion lens) microscopic magnification [18] . Statistical analysis was done for the previous parameters using the (t) test for data evaluation and for comparison between the experimental groups. Data was expressed as means ± St nd rd Devi tion (SD) P v ue ≤ 5 w s onsidered statistically significant.