Journal of Chemical, Biological and Physical ...

4 downloads 0 Views 216KB Size Report
Nov 2, 2014 - 3 Department of Medical Laboratory Science, Madonna University, Elele, Nigeria. Received: 30 September 2014; Revised: 26 October 2014; ...

JCBPS; Section B; Nov . 2014 – Jan. 2015, Vol. 5, No. 1; 311-319.

E- ISSN: 2249 –1929

Journal of Chemical, Biological and Physical Sciences An International Peer Review E-3 Journal of Sciences Available online atwww.jcbsc.org

Section B: Biological Sciences CODEN ( USA): JCBPAT

Research Notes

Effect of methanolic root extract of napoleona imperialis on some haematological parameters in albino rats. Okon Effiom ETIM 1 *, Remy Ukachukwu DURU2 , A.A. MARIE3 , and Gladys Ozisiaka OMETERE4 1*, 4

Department of Biochemistry, Madonna University, Elele, Nigeria.

2 3

Department of Chemistry, Madonna University, Elele, Nigeria.

Department of Medical Laboratory Science, Madonna University, Elele, Nigeria.

Received: 30 September 2014; Revised: 26 October 2014; Accepted: 02 November 2014

Abstract: Napoleona imperial is is an evergreen non-timber plant that has wound healing and antihypertensive effect. This study is aimed at investigating the effect of methanolic root extract of napoleonaimperialis on some heamatological parameters of albino rats. The experimental animals were grouped into five groups with 6 animals each. Group 1(control)received 0.9% normal saline while group 2, 3, 4 and 5 received 400mg/kg, 800mg/kg, 1000mg/kg, 1200mg/kg of methanolic root extract of napoleona imperialis respectively for 21days. White blood cell (WBC), red blood cell (RBC), haemoglobin (HGB), packed cell volume (PCV), Lymphocyte count, Neutrophil count, Mean cell volume (MCV), Mean cell haemoglobin (MCH), Mean cell Haemoglobin Concentration (MCHC) were determined using standard methods. The result shows a significant difference (p0.05) in TWBC, LYM, NEU and MCHC of groups 3, 4 and 5 when compared to control and other groups. This result therefore suggests that methanolic extract of Napoleona imperialis at certain concentration boost haematological parameters like haemoglobin level and packed cell volume. Keywords: Napoleona imperialis. Haematologicalparameters, albino rats, wound healing and antihypertensive effect. 311

J. Chem. Bio. Phy. Sci. Sec. B, Nov 2014 – Jan. 2015; Vol.5, No.1; 311-319.

Effects …

Okon Effiom et al.

INTRODUCTION The indigenous medicinal plants in Nigeria form an important component of the natural wealth of the country. Most of these plants have been used indiscriminately by many local populations for managing various diseased states without actually knowing how relief is brought about or its safety and toxicity risk. Napoleonaimperialis is an evergreen non-timber plant that grows abundantly in bush fallows, secondary bushes and marginal lands in most of the tropical humid zones of West Africa. The plant belongs to the family known as the lecythidaeceae, along with the cannon ball tree (corrupitaguianensis), which grows in most regions of Nigeria1 . Haemtological studies are useful in the diagnosis of many diseases as well as investigation of the extent of damage to blood2 . The examination of blood gives the opportunity to investigate the presence of several metabolites and other constituents in the body of animals and it plays a vital role in the physiological, nutrition and pathological status of an organism3 ,4 . It has been reported that, different parts of N.imperialis are used for different purposes in the region including mulching and fodder (leaves and twigs); and firewood, chewing sticks and ethnomedicine (stem and root)5,6 ,. Therefore this study attempt to investigate the effect of administration of methanolic extract of Napoleona imperialis roots on hematological parameters using albino rats. MATERIALS AND METHODS Materials: The plant Napoleona imperialis was obtained from Mgbirichi community in Imo state and identified at the herbarium of University of Nigeria, Nsukka. Standard laboratory apparatus were provided including centrifuge, electric water bath (manual).electronic weighing balance (SCONT PRO), rotary evaporator and spectrophotometer (UNISPEC 23D). Methanol, normal saline and all other chemicals used were of analytical grade and purchased from Sigma Aldrich, USA. METHODS Extraction and Preparation of plant Materials: The roots of Napoleona imperialis were cut into pieces, washed, dried and grounded for extraction. Themethanolic extract of Napoleona imperialis was obtained using 80% of methanol. The mixture was concentrated with thermostatic water bath at 50o C. Experimental Design: Thirty (30) albino rats were used for this assay and grouped into five (5) groups of six (6) each according to body weight (80-150g). Stock solution (200mg/ml of normal saline) of napoleona imperialis root extract was prepared and administered thus for 21 days; • Group 1(control) received 0.9% normal saline • Group 2 received 400mg/kg b.w of N.imperialis root extract. • Group 3 received 800mg/kg b.w of N.imperialis root extract. • Group 4received 10000mg/kg b.w of N.imperialis root extract. • Group 5 received 1200mg/kg b.w of N.imperialis root extract.

After days of administration, rats were anesthetized with chloroform, sacrificed and blood collected by cardiac puncture and stored in an EDTA sample bottles for haematological assay within 12 hours. 312

J. Chem. Bio. Phy. Sci. Sec. B, Nov 2014 – Jan. 2015; Vol.5, No.1; 311-319.

Effects …

Okon Effiom et al.

Determination of Packed cell volume (Microhaemotocrit Method): The packed cell volume is that proportion of whole blood occupied by red cells, expressed as a ration (litre/ litre). Anticoagulated blood in a glass capillary of specified length bore size, and wall- thickness is centrifuged in a microhaemocrit centrifuge at RCF 12000-15000 rpm for 3-5minutes to obtain constant packing of the red cells. The PCV value is read from a scale of a microhaemocrit reader or calculated by dividing the height of the red cell column by the height of the total column of blood7 . Determination of Hemoglobin concentration (Haemiglobincyanide (HiCN)/ Cyanmethaemo globin technique): Whole blood is diluted 1 in 201 in a modified Drabkin’s solution which contains potassium ferricyanide and potassium cyanide. The red cells are haemolysed and the haemoglobin is oxidized by the ferricyanide to methaemoglobin. This is converted by the cyanide to stable haemoglobincyanide (HiCN). The haemoglobin concentration is directly proportional to the absorbance of the HiCN solution when read in spectrophotometer at wavelength 540nm or in a filter colorimeter using a yellow-green filter7 . Calculation:

Hemoglobin Concentration

= Absorbance of sample× Conc. of standard Absorbance of standard

Determination of Total White Cell count (Haemocytometry): Whole blood is diluted 1 in 20 in an acid reagent (Torks solution) which haemolyses the red cells, leaving the white cells to be counted. White cells are counted microscopically using an Improved Neubauer ruled counting chamber (haemocytometer) and the number of WBC’s per litre of blood calculated7 . Calculation:

Number of white cells counted =N White cell count= [(N÷2) ÷ 10] ×109 /L

Determination of Differential leukocyte count and Blood cells morphology (Stained thin film microscopy): When a well-made thin blood film is stained with leishman stain which is a Romanowsky stain. The basic part of the stain (methylene blue), stains the acidic component of the cell (the nucleus), while the acidic part of the stain (eosin) stains the basic components of the cell (the cytoplasm). STATISTICAL ANALYSIS Data obtained from this study were analyzed using the statistical package for social sciences (SPSS) version 18.0 for windows. Analysis of variance (ANOVA) were used to compare means, and values were considered significant at p0.05).

315

J. Chem. Bio. Phy. Sci. Sec. B, Nov 2014 – Jan. 2015; Vol.5, No.1; 311-319.

Effects …

Okon Effiom et al.

Figure V: Mean Neutrophil Count in Napoleonaimperialis extracts Fed Albino Rats. Data represented as Mean ± SEM. Mean Cell Volume in Napoleona imperialis extracts Fed Albino Rats: As shown in Figure VI below, rats administered 400mg/kg extract had significantly higher mean cell volume (76.1 ± 3.94mg/dl) compared to the normal control (59.8 ± 1.56mg/dl)and those administered 800,1000,1200mg/kg extract (P˂0.05).

Figure VI: Mean Cell Volume in Napoleona imperialis extracts Fed Albino Rats. Data represented as Mean ± SEM. Mean Cell Haemoglobin in Napoleonaimperialis extracts Fed Albino Rats: As shown in Figure VII below, rats administered 1200mg/kg extract had significantly higher mean cell haemoglobin (22.2 ± 1.6 mg /dl)compared to the normal control (15.7 ± 0.66mg/dl) and those administered 400,800,1000mg/kg extract. However, the mean difference was not statistically significant (P>0.05).

316

J. Chem. Bio. Phy. Sci. Sec. B, Nov 2014 – Jan. 2015; Vol.5, No.1; 311-319.

Effects …

Okon Effiom et al.

Figure VII: Mean Cell Haemoglobin in Napoleonaimperialis extracts Fed Albino Rats. Data represented as Mean ± SEM. Mean Cell Haemoglobin Concentration in Napoleona imperialis extracts Fed Albino Rats: As shown in Figure VIII below, rats administered 1000mg/kg extract had significantly higher mean cell haemoglobin concentration (33.2 ± 4.02mg/dl)compared to the normal control (26.5 ± 1.50mg/dl) and those administered 400,800,1200mg/kg extract. However, the mean difference was not statistically significant (P>0.05).

Figure VIII: Mean Cell Haemoglobin Concentration in Napoleona imperialis extracts Fed Albino Rats. Data represented as Mean ± SEM. DISCUSSION Haemoglobin is the iron-containing oxygen-transport metalloprotein in the red blood cells of all vertebrates8 with the exception of the fish family, channichthyldae 9 as well as tissues of invertebrates. Haemoglobin has the physiological function of transporting oxygen to tissues of the animal for oxidation of ingested food so as to release energy for the other body functions as well as transport carbon dioxide out of the body of animals10-13 . As ahown in Figure I, rats administered 1200mg/kg extract had significantly higher (p