The Effect of Cajanus Cajan Seeds Extraction on Some Haematological Parameters Among Haemorrhagical Anaemic Rats in Khartoum–Sudan Some Haematological Parameters Among Haemorrhagical Anaemic Rats in Khartoum–Sudan.

Background: Worldwide different plants were studied for their role in the treatment and correction of different types of anaemia’s. The Medicinal and Aromatic Plants Research Institute (MAPRI) of the National Centre for Research (NCR) plays major role in medicinal and aromatic plants investigation, determination of therapeutic properties and documented in Sudan. Objectives : This study used to determine the effect of Cajanus Cajan seeds extract on the some haematological parameters among hemorrhagic anaemia rats, Khartoum –Sudan. Methods : Experimental randomised controlled trial study conducted among 24 healthy Wistar albino rats with both sex, ages (6weeks) and weighing between (75-130g), they were feed with standard laboratory chow. they divided into four groups each one consists of six rats. Result: Dose group(800 mg/kg/day) represent highly significant increases in haemoglobin in W0&W1 P value < 0.001,and W0&W3 p value is 0.0002.while the dose of 400mg/kg/day significant increase in W0&W1 p value is< 0.003 and W0&W3 p<0.00004, in the dose of 200mg/kg/day represent also significant increase at W0&W1 p<0.005 and in W0&W3p<0.007 respectively, when compared to control values(Group1). Conclusion : This study revealed that there are increased in some haematological parameters and lend support to the use of ethanol seeds extract of in treatment of hemorrhagic anaemia.

Plant is an important source of medicine and plays a key role in world health [6]. Medicinal herbs or plants have been known to be a vital potential source of therapeutics or curative aids. This includes the use of medicinal plants not only for the treatment of diseases but also as potential material for maintaining good health and conditions. majority of the used drugs contain plant extracts. Some contain active ingredients (bioactive components or substances) obtained from plants. Through recent researches, plant-derived drugs were discovered from the study of curative, therapeutic, traditional cures and most especially the folk knowledge of local people and some of these claims and believe of people are irreplaceable despite the recent advancement in science and technology.

The Role of Medicinal Plants in Traditional Healing
The pharmacological treatment of disease began long ago with the use of herbs. Methods of folk healing all over the world commonly used herbs as part of their tradition, It is also a role of the traditionally-held belief that the synergistic mixture of several active principles in some herbal preparations is responsible for their beneficial effects [7]. Methods of traditional healing throughout the world usually used herbs as part of their tradition.
The introduction of plant derived drugs in recent medicine has been linked to the uses of plant derived materials as an indigenous cure in traditional system of medicine [8]. Some of the plants have been found to possess significant antibacterial, antifungal, anticancer, antidiuretic, anti-inflammatory and anti-diabetic properties [9][10][11][12].

Traditional Uses of Some Plants
Fabaceae is the third largest family of flowering plants with more than 18,000 described species including herbs, shrubs, trees and vines distributed throughout the world, especially in tropics rain forest and widespread in Sudan [15] [17]. Pigeon pea is an essential component of many people's diet in Asia, Africa and South America [18] It has been used to a attain the requirements of food, forage, feed, and therapeutics.
Conventional consumption of seed is as a green vegetable and dry pulse, but young green pods, shoots, and leaves are also edible.
Bioactive components in pulse seeds are getting increased attention in recent years.

Physiological Activities of Pigeon Pea Seeds
Antihyperglycemic & Lipid Peroxidation: The indirect effect of total phenolics with increased antioxidant activity leading to the inhibitory potential of carbohydrate digesting enzymes and a positive correlation between consumption of germinated pigeon pea and controlling hyperglycemia. Reduction in lipid peroxidation and reduced fasting blood glucose level in diabetic rats [19]. The anti-hyperglycemic activity was found after administration of a single dose of unroasted seeds (60% and 80%)and administration to normal as well as alloxanized mice, significant reduction in the glucose level in the blood after 1-2 hours and significant increase at 3hours [20]. Anti-Inflammatory Activity: Investigational study on the effect of 50% ethanol extracts of pigeon pea, as well as its main component, cyanidin-3-monoglucoside, an anthocyanin, on DNA damage, the activity of antioxidant enzymes, and free radical scavenging capacity in hydrogen peroxide (H2O2)-treated RAW264.7 macrophages. 50% ethanol extracts of pigeon pea and cyanidin-3-monoglucoside suppressed the production of inflammatory cytokines, including TNF-α, IL-1β, and IL-6, in these macrophages [22]. Also Cajaninstilbene acid (CSA) only found in pigeon pea and extracted from its leaves along with its synthesized derivatives revealed strong inhibition activity on the release of nitric oxide(NO) and inflammatory factor TNF-α and IL-6 in lipopolysaccharides (LPS)-stimulated mice macrophages [23].
Anti-Sickling Activity: Anti-sickling experiments depend on the estimation of free phenylalanine in the methanol (watersoluble) extract of the white variety seeds. The presence of phenylalanine amino acid alone could account for about 70% of the antisickling potency of Cajanus cajan seed extract [24].

Ethanolic extraction of Cajanus Cajan Seeds
The Cajanus Cajan fresh seeds crushed to powder, dried and prepared plants material(400g) was suspended in 1 litre of 80% ethanol for( 24hr) this suspension was re suspended in an equal volume of 80% ethanol for (48hr) and filtrated again the filtrates were pooled and the solvents were evaporate in a Rotavapor (Switzerland,BUCHI00I) or under reduced pressure [4].

Haematological Methods
Sample Collection: In EDTA blood sample these techniques were performed according to an automated haematology analyser

Procedures
Blood sample were collected from each group four times in week zero immediately after the bleeds, then after the first, second and third weeks. examined for determination of haemoglobin concentration, red blood cell count, total white blood cells count, packed cell volume, mean corpuscular volume, mean corpuscular haemoglobin, and mean corpuscular haemoglobin concentration.

Statistical Analysis
Statistical analysis was performed using package for social

Haematological Results
There are significant (p<0.05) progressive blood parameter (HB, PCV, RBCs) related to the number of doses. After 3 weeks the result analyzed statistically by SPSS and Excel and expressed as follow: Table 4-1 showed the Mean, Mode and SD for (RBCs, HB,), Table 4-2 showed Mean, Mode and SD for(PCV,PLT) ( Table 1). Table 4-3 showed showed Mean, Mode, SD of (MCV, MCH). Table 4-4 showed Mean, Mode, SD of MCHC (Table 2). Table 4

Discussion
The result of the present study showed that the body mechanism of the bled rat responded to about blood loss by rapid generation of RBCS with in 3weeks and increase in HB, PCV by administration of ethanolic extract of cajanus cajan in significant result(p<0.05).
Administration of alcoholic extraction of cajanus cajan increase the recovery of treated rats from effect of bleeding. The lowest administration dose 200mg/kg reduced the recovery time of most blood parameters from about 3weeks in the bled control to 1week. in addition, the extent of recovery was dose related with highest dose of 800mg/kg effecting the highest change. The observed elevation of mean cell volume in bled rats by week1 is indicative of anaemia due to acute blood loss.

Conclusion
Pigeon pea is a perennial tropical crop primarily grown in Asia and Africa, and its seeds are consumed as a rich source of protein and carbohydrates both in fresh and dried forms. This crop has been successfully grown in some southeastern states but still considered as a novel pulse here in the US. Majority of the work focused on its non-consumable parts like leaves, stems, and roots. In this study we used the seeds extraction of Cajanus Cajan to work on it. Literature studies indicate that pigeon pea has the potential to prevent and treat many human diseases such as bronchitis, pneumonia, measles, hepatitis, yellow fever, ulcers, diabetes, and certain forms of cancer.
Along with protein and fiber, it has a decent number of healthpromoting phytochemicals. Important phytochemicals found in pigeon pea seeds are phenolic acids, flavonoids, tannins, saponins, and phytic acid. These minor components predominately exhibit and are capable of antioxidant, antidiabetic, and anti-inflammatory activities. Seeds are the edible and non-perishable part of this crop with the feasibility of addition in food products. Exploring the nonstarch polysaccharide fractions and functional properties of the pigeon pea flour can make it a gluten-free substitute for cereals.
The overall results of this study lend support to the folkloric use of ethanol seed extract of Cajanus Cajan in treatment of haemorrhagic anaemia.