Haemopoietic Effect of Telfairia Occidentalis Diet Preparation in Albino Rats

Telfairia occidentalis, a member of the cucurbitaceae family popularly known as fluted pumpkin is highly reputed in traditional medicine practice in Nigeria (Akpapunan et al., 1995). It is commonly called Ugu in most parts of Nigeria with different parts of the plant being employed for various disease conditions ranging from convulsion, Anaemia and Malaria

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. The leaves are rich in essential and non-essential amino acids, vitamins and minerals (Fasuyi, 2006).
Aqueous extracts of T. occidentalis leaves is used locally in Nigeria as antimalaria and haematinic agent.

The aqueous extract of the leaves has been reported to possess significant enhancement of haematological parameters (erythrocyte count, packed cell volume, haemoglobin concentrate and white blood cell count), antiparastic effect on Plasmodium berghei infected mice as well as hypoglycaemic activity (Salman et al., 2008). In view of the increasing rate at which herbal preparations are used traditionally as medicine, there is the need to investigate the clinical implications of herbal preparations.

Earlier reports on the enhancement of haematological parameters of aqueous extract of the leaves of T. occidentalis prompted this study. The study was aimed at evaluating the possible clinical implications of T. occidentalis as blood tonic in order to proffer support or question the feasibility of developing a cost-effective and efficacious management for Anaemia using herbs especially in poor resourced economy, since it is known that 60% of the Global population does not have readily available access to these modern drugs and medications thereby leading to an increased mortality rate under anaemic conditions. Hence the need arose to seek cheaper available alternatives to help alleviate these disease and other related ones.
Justification
Anaemia have a high prevalence in the society, despite the high consumption level of this the leaves which has been claimed to reduce the prevalence of this condition. However, more efforts are needed to ensure that the claim is justified and to make the plant easy accessible by the population.

Aim/Objectives
T. occidentalis have been claimed to boost low blood volume and therefore based on this claim, this study is aimed at determining the validity of this claim such as the haematinic agents contained in these leaves it is also intended that the study will provide more insights that will necessitate further studies on this plant.

CHAPTER TWO
LITERATURE REVIEW

HAEMOPOETIC ACTIVITY
Haemopoesis is the formation of blood cellular components. All cellular blood components are derived from haemopoetic stem cells. In a healthy adult person, approximately 1011 – 1012 new blood cells are produced daily in order to maintain steady state levels in the peripheral circulation.

HAEMOPOIETIC STEM CELL (HSCs)
Haemopoietic stem cells (HSCs) reside in the medulla of the bone (bone marrow) and have the unique ability to give rise to all of the different mature blood cell types and tissues. HSCs are self-renewing cells. When they proliferate, at least some of their daughter cells remain as HSCs, so the pool of stem cells does not become depleted. This phenomenon is called asymmetric division (Morrison and Judith, 2006). The other daughters of HSCs (Myeloid and Lymphoid progenitor cells), however can commit to any of the alternative differentiation pathways that lead to the production of one or more specific types of blood cells, but cannot self-renew. The pool of progenitors is heterogeneous and can be divided into two groups, long-term self-renewing HSC and only transiently self-renewing HSC, also called short-terms (Morrison and Weissman, 1994).
This is one of the main vital processes in the body. All blood cells are divided into three lineages.
• Erythroid cells are the oxygen carrying red blood cells. Both reticulocytes and erythrocytes are functional and are released into the blood.
• Lymphocytes are the cornerstone of the adaptive immune system. They are derived from common lymphoid progrenitors. The lymphoid linage is primarily composed of T-cells and B-cells (types of white blood cells). This is lymphopoiesis.
• Myelocytes, which include granulocytes, megakaryocytes and macrophages and are derived from common myeloid progenitors are involved in such diverse rates as innate immunity, adaptive immunity and blood clotting. This is myelopoiesis.
However, when there is disorder of erythroid cells, it results in the reduction of oxygen carrying capacity of the blood in which haemoglobin level falls below normal.

ANAEMIA
Anaemia may be defined as a condition in which oxygen carrying capacity of blood is reduced. It is always a sign of an underlying disease, and never a diagnosis. Patients with anaemia have similar symptoms irrespective of the cause. Such symptoms may include fatigue, dizziness, breathlessness and headache. Causes of Anaemia includes;
• Increased haemolysis
• Decreased erythropoesis
• Excessive blood loss

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However, several conditions are associated with decreased red blood cell level which include;
• Bleeding lesion in the digestive tract (gastric ulcer)
• Heavy bleeding during surgery trauma.
• Iron deficiency, due to insufficient dietary intake or absorption from the digestive tract.
• Chronic renal failure.

TYPES OF ANAEMIA
MEGALOBLASTIC ANAEMIA
This group of anaemias is as a result of defective DNA synthesis leading to delayed maturation of the nucleus of erythroblasts as compared to the maturation of their cytoplasm. The bone marrow shows large erythroblasts (megaloblasts) with immature nucleus and normal haemoglobinisation of the cytoplasm.
The defect in DNA synthesis is most commonly due to deficiency of vitamin B12 or folic acid, or sometimes due to abnormalities of metabolism of these substances. Both vitamin B12 and folic acid play an essential role in cell metabolism. The act as coenzymes in the synthesis of DNA (Ochei and kohatkar, 2004).

HAEMOLYTIC ANAEMIAS
Haemolytic anaemias are those anaemias that result from increased destruction of red blood cells. In severe haemolytic anaemias the red blood cells survive only for a few days instead of a normal life span of 120 days. The causes of this increased haemolysis can be hereditary membrane disorders such as hereditary sphelocytosis, hereditary elliptocytosis or ovalecytosis, Metabolism as in the case of Glucose 6 phophate dehydrogenase (G6PD) deficiency, Haemoglobin such as HbS, HbC, etc. however, haemolytic anaemias could be acquired or immune secondary.

APLASTIC ANAEMIA
Aplastic anaemia is a disorder of haematopoiesis characterized by pancytopaemia in the peripheral blood and a hypoplastic bone marrow with no signs of granulomatous disease or malignancy in the bone marrow.

SCIENTIFIC PROFILE OF TELFAIRIA OCCIDENTALIS
Telfairia occidentalis is a tropical vine grown in West Africa as a leaf vegetable and for its edible seeds common names for the plant include, fluted gourd, fluted pumpkin, and ugu. Telfairia occidentalis is a member of the curcurbitaceae family and is indigenous to southern Nigeria (Akoroa, 1990).
The fluted gourd grows in many nations of West Africa but is mainly cultivated in Nigeria, used primarily in soups and herbal medicines.

Although the fruit is inedible, the seeds produced by the gourd are high protein and fat, and can therefore contribute to a well-balanced diet. The plant is a drought-tolerant, dioecious perennial that is usually grown trellised. Telfairia occidentalis is traditionally used by an estimated 30 to 35 million indigenous people in Nigeria including Efik, Ibibio, and Urhobo (Akoroda, 1990).
However, it was predominantly used by the Igbo tribe, who continue to cultivate the gourd for food sources and traditional medicines (Okoli et al., 1983). A recurring subject in the Igbo’s folklore, the fluted gourd is noted to have healing properties and was used as a blood tonic to be administered to the weak or ill (Akoroda, 1990).

STRUCTURE/PSYCHOLOGY
The fluted gourd fruit is quite large; one study documenting a range of 16-105 centimeters in length, and an average of 9 centimeters in diameter (Okoli, 1983). The same study found the seed count in larger gourds to reach upwards of 196 per fruit, typically measuring between 3.4 and 4.9 centimeters in length. In both the pistillate and staminate varieties, Telfairia occidentalis flowers grow in sets of five, with creamy-white and dark red petals, constrasting with the light green colour of the fruit when young and yellow when ripe.

NUTRITIONAL CONTENTS
Considered in “oil seed”, the fluted gourd is high in oil (30%) (Akoroda, 1990). Shoots of Telfairia occidentalis contain high level of potassium and iron, while seeds are composed of 27% crude proteins and 53% fats (Aiyelaagbe and Kintomo, 2002).
The young shoots and leaves of the female plant are the main ingredients of a Nigeria soup. The large (up to 5cm), dark-red seed is rich in fat and protein and can be eaten whole, ground into power for many kinds of soups, or made into a fermented porridge.

USES
The edible seeds of Telfairia occidentalis can be boiled and eaten whole or fermented and added to “ogili” (Badifu, 1993).
The fluted gourd has been traditionally utilized by indigenous tribes as a blood tonic, likely due to it’s high protein content. Flour produced by the seeds can be used for high-protein breads, although more research is needed to fulfill this potential food source (Giami, 2003).
Furthermore, the shoots and leaves can be consumed as vegetables.

PRACTICAL INFORMATION
The roots of Telfairia occidentalis are locally known to be a potent poison to humans and therefore should not be consumed (Akoroda, 1990). However, the seeds of the fluted gourd are very high in oils, and have potential to become a staple, high-protein oil food source in Nigeria. One study conducted to investigate levels of protein pre and post germination, found that germination significantly increased albumin and globulin protein factors, which has been shown to increase levels of lysine in a plant. Furthermore, male plants mature on average 11days before female, and continue to produce flowers after female plants stop producing flowers (Akoroda, 1990).

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SCIENTIFIC CLASSIFICATION OF TELFAIRIA OCCIDENTALIS
The plant is classified thus:
Kingdom Plantae
Division magnoliophyta (flowering plant)
Class magnoliopsida (dicot)
Order cucurbitales
Family cucurbitaceae
Genus Telfairia
Specie T. occidentalis

PREVIOUS INVESTIGATIONS

Phytochemical Compositions of Telfairia Occidentalis
Phytochemical screening of the aqueous extract of telfairia occidentalis leaves showed the presence of alkaloids, cardenolids, anthraquinones, saponins, flavonoids and tannins (Igoli et al., 2005).

phytochemical constituents of aqueous extract of Telfairia occidentalis leaves.

Phytochemical Dry leaves extract Ethanolic extract
Alkaloids ++
Anthraquinones + +
Saponins + ++
Flavonoids
Tannins

Key: (Absence); + (presence).

The phytochemical analysis carried out on the dry leaves water extract and ethanolic extract showed the presence of some bioactive compounds in the plant. In the two forms of extract, five bioactive constituents were tested for, out of which only two were present in the two extractions (table 1). Analysis of alkaloids in the two extracts found that alkaloids were only present in ethanolic extracts while absent in the dried leaf water extract. The main reason that can be adduced for this is the mode of extraction. On this premise it will be advisable to extract the leaf of T. occidentalis with ethanol in an attempt to exploit it’s detoxifying and antihypertensive properties since alkaloids is known for these purpose (Trease and Evans, 1997).
However, flavonoids was found to be negative in the two found to be negative in the two extracts of the leaf it was not really surprising because the sole greenish appearance of the leafy vegetable substantiates the fact that flavonoids contribute to the brilliant multi colour for most plants (said by sofowora, 1993).

TELFAIRIA OCCIDENTALIS AS FOOD PLANT
Telfairia occidentalis (fluted pumpkin) are used primarily in soup. The seeds produced by the pumpkin are high in protein and fat, and can therefore contribute to a well-balanced diet. It can be eaten whole, ground into powder for many kinds of soups. Shoots of T. occidentalis contains high level of potassium and iron (Aiyelaagbe and Kintomo, 2002).

MEDICAL USES OF T. OCCIDENTALIS
Although the plant is mainly cultivated as food, it has continued to be an important medicinal plant. Much of it’s recent spread into new areas may likely be attributed to its medicinal value. A wide variety of claims have been made for its medicinal efficacy as a treatment for numerous ailments ranging from its ability to enhance haematological parameters to cure for convulsion and malaria (Fasuyi, 2006). In view of the reputed efficacies of this vegetable plant, studies have investigated it’s phytochemical constituents in an attempt to establish its most active form and antiparasitic activities against plasmodium berghei.

The effect of the aqueous extract of T. occidentalis on the ability of artesunate to clear parasitaemia due to P. berghei is presented below:
Evaluation of the haematological parameters showed an insignificant enhancement of the PCV by T. occidentalis on the 3rd day of administration in the diseased state. PCV of 33.3+ 1.445% was obtained for the control group in the presence of infection while 37 + 0.58% was obtained for the aqueous extract of T. occidentalis. Similarly, 42.5 + 0.96% was obtained in the absence of infection while 44.5 + 0.65% was obtained after the administration of the extract. However, the co-administration of artesunate with the aqueous extract gave a significant enhancement of PCV when compared with artesunate and extract alone at day 3 (P < 0.01).
Similar trend was also observed with haemoglobin concentration, RBC, WBC, platelet count, neutrophils and eosinophils (Okokon et al., 2009).

HAEMOPOIETIC PARAMETERS UNDER INVESTIGATION

PACKED CELL VOLUME (PCV) HAEMATOCRIT
PCV also known as haematocrit or erythrocyte volume fraction is that proportion of blood volume occupied by the red blood cells. Its unit measurement is in percentage (%) it is normally 45% for men and 40% for women (Purves et al., 2004). It is considered an integral part of person’s complete blood count results, along with haemoglobin concentration, white blood cell count, and platelet count. Anaemia refers to an abnormally low hematocrit, as opposed to polycythemia, which refers to an abnormally high hematocrit. Both are potentially life threatening disorders.

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Etymology
The term hematocrit comes from the Ancient Greek word haima (“blood”) and krites (‘’judge’’). It was coined by Magnus Blixat Uppsala in 1891 as haematokrit (Stanley, 1981).
The packed cell volume can be determined by centrifuging heparinized blood in a capillary tube (also known as microhaematocrit tube) at 12, 000rpm for five minutes. This separates the blood into layers. The volume of packed red blood cells divided by the total volume of the blood sample gives the PCV. Since a tube is used, this can be calculated by measuring the lengths of the layers.
With modern lab equipment, the haematocrit is calculated by an Automated analyzer and not directly measured. It is determined by the mean cell volume. The haematocrit is slightly more accurate as PCV includes small amounts of blood plasma trapped between the red cells. An estimated hematocrit as percentage may be derived by tripling the haemoglobin concentration in g/dl and dropping the units (Jelkmann, 2004).

HAMOGLOBIN CONCENTRATION
Haemoglobin is an iron-containing compound found in the red blood cells, which transports oxygen around the body. Measuring the concentration of haemoglobin in the blood can help to detect anaemia and its severity and to monitor an anaemic patient’s response to treatment. Monitoring the haemogblobin level (or PCV) is also required when patients with HIV disease are being treated with drugs such as Azidothymidine (AZT). The test is also performed to check the haemoglobin level of a blood donor prior to donating blood. In district laboratories, depending on available facilities and resources, haemoglobin is measured photometrically or estimated using a visual comparative technique. Haemogblobin values are expressed in grams per litre (g/l) or grams per deciliter (g/dl) (Cheesbrough, 2006).

TOTAL WHITE BLOOD CELL (WBC) COUNT
White blood cells also known as leukocytes are the cells of the immune system that are involved in defending the body against both infectious disease and foreign invaders (Lafleur-Brooks, 2008).

There are five types of leulocytes such as neutrophil, eosinophil, Basophil, monocyte and lymphocyte. All leukocytes are produced and derived from a multipotent cell in the bone marrow as a haemopoietic stem cell. They live for about three to four days in the body. Leukocytes are found throughout the body, including the blood and lymphatic system (Maton et al., 1997).
The number of leukocytes in the blood is often an indicator of disease. The value is expressed in times ten raise to power nine per litre (x109/l).

Etymology
The name “white blood cell” derives from the physical appearance of a blood sample after centrifugation. White cells found in the buff, a thin, typically white layer of nucleated cells between the sedimented red blood cells and the blood plasma. The scientific term leukocyte directly reflects its description. It is derived from the Greek word leuko meaning “white” and kytos meaning “hollow vessel,” with cyte translated as “cell” in modern usage. Buffy coat may sometimes be green if there are large amounts of neutrophils in the sample, due to the heme-containing enzyme myeloperoxidase that they produce (Gartner and Hiatt, 2007).

OTHER PLANTS WITH RELATED MEDICAL IMPORTANCE
Many vegetables that are of medical importance exists in nature. A wide variety of these plants have been investigated and others are yet to be discovered.
The haematinic properties of the aqueous extract of hibiscus cannbinus (Kenaf) leaves was studied on phenylhydrazine-induced anaemic rats (Gabriel, 2005).
The orally administered extract induced a significant decrease (P<0.05) in the blood parameters indicating anaemia and also resulted to a significant increase (P < 0.05) in the MCH, MCV, which are indicators of increase in the RBC count, Hb concentration, and PCV which had been originally decreased by phenylhydrazine administration within one week of treatment.

Based on these ethno medical uses, this study is aimed at determining the validity of some of these claims, especially the haematinic properties. It is also intended that the work will also provide some insights that will necessitate further studies on this plant.

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