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Effect of Gestational Oral Ricinus communis Oil Ingestion on Haematological Parameters in Wistar Rats

Salami S.A

Department of Physiology, Lagos State University College of Medicine, Ikeja

Abstract: Objective: Haematological effect associated with Ricinus communis oil (RCO) ingestion at different gestation periods was investigated in wistar rats. Methods: 30 pregnant rats were randomly shared into 5 groups of six rats each. Group 1 (control) received distilled water while groups 2-5 received 950 mg/kg BW RCO orally between gestation days (GD) 1-7, 7-14, 14-21, and 1-21 respectively. Maternal haematological parameters were determined by conventional methods and data were analyzed using graph pad prism software. Statistical significance was taken at p<0.05. Results: There were significant increases in packed cell volume (PCV), haemoglobin concentration (Hb) and red blood cell count (RBC) in GD 14-21 treated rats as compared to control. However mean corpuscular volume (MCV) and mean corpuscular haemoglobin (MCH) decreased significantly. GD 1-7 treated rats showed a reduction in PCVwhen compared to control. Percentage variations/changes in leucocyte cells values were all not statistically different from control values. Conclusion: Oral RCO exposure at different gestation periods has no significant effect on differential leucocytes cells count but appears to reduce and enhance erythropoiesis at early and late gestation period respectively.

Keywords: ricinus communis oil, gestation days, haematological parameters, pregnant wistar rats

Citation: Salami S.A, Effect of Gestational Oral Ricinus communis Oil Ingestion on Haematological Parameters in Wistar Rats. American Journal of Biomedical Research, 5 (1). 14-16. January 2017. doi:10.12691/ajbr-5-1-3

1. Introduction

Issues of morbidity/mortality in pregnancy have been endemic in developing world particularly sub-Saharan Africa with associated causes often multifaceted [1,2,3,4]. One often reported cause especially in rural communities are unverified herbal products use by pregnant women [5,6]. Ricinus communis oil (RCO) despite being reported contra-indicated in pregnancy by few studies; other studies still reported on its preferred use as labour inducer by some traditional birth attendants [7,8,9]. In Nigeria where this study was carried out, traditional birth attendants still rely on the use of local herbs and concoction like RCO during pregnancy. Apart from exposure through traditional birth attendants, pregnant women are also inadvertently exposed to RCO during different stages of pregnancy due to its accessibility and availability oftentimes as over the counter drug. We have previously reported on the effects of RCO on biochemical and hormonal parameters in pregnant rats [10]. Furthermore, we also reported on the generational reproductive implication of maternal exposure to RCO in offspring of exposed mothers [11,12]. However, literature search shows specifically nothing on effects of RCO on haematological parameters of pregnant rats during different gestation periods. This study investigates the gestational haematological effect after RCO consumption in Wistar rats.

2. Materials and Methods

2.1. Animals, Ethical Approval, Experimental Design and Treatment

Animals used were sourced from the Lagos state University College of Medicine and allowed access to standard rat pellets (Ladokun feeds) and water ad libitum. Guidelines for animal care and use were strictly adhered to and ethical approval given by the college animal house committee (LASUCOM/EAC/27). Thirty mature nulliparous female (220-250 g) albino rats (12 weeks) with normal estrous cycle were used. Positive mating was confirmed by the presence of a sperm positive vaginal smear or copulatory plug and day after which either of this was found was taken as gestational day 0 as we have previously reported [12]. 950 mg/kg BW of RCO (recommended therapeutic dose in humans, (Drug store.com, Inc. 2004) [13] was administered orally to pregnant wistar rats (6 per group) at gestation days 1-7, 7-14, 14-21 and 1-21 respectively (groups 2-5). Control group received distilled water (group 1). The treatments to groups were chosen to target all the critical periods of gestation (i.e. early, mid, late and entire gestation periods).

2.2. Preparation of RCO

Ricinus communis seeds were air dried to a constant weight and then pulverized. The pulverized seeds (2 kg) were extracted by cold extraction using 5 litres of methanol. RCO obtained by filtration was evaporated of the solvent in a rotatory evaporator at 37°C [10].

2.3. Collection of Blood from Pregnant Rats and Determination of Haematological Parameters

For haematological studies, blood samples (2ml) were collected in heparinized bottles via the orbital sinus under penobarbitone anesthesia. Blood samples were subsequently analyzed using automated blood analyzer H1Z Hema-screen 13 by Hospitex Diagnostic, San Pierro Quaracchi, Firenze, Italy. Red blood cell count (rbc) hemoglobin concentration (Hb), packed cell volume (pcv), mean corpuscular volume (mcv), mean corpuscular hemoglobin (mch), mean corpuscular hemoglobin concentration (mchc) and differential leucocytes counts (lymphocytes, neutrophils, eosinophils and monocytes) were all determined and recorded.

2.4. Statistical Analysis

Mean values and standard error of mean (Mean ± SEM) were calculated. The test of significance between two groups was by Students T test and for more than two groups by the analysis of variance (ANOVA) using prism graph pad statistical software version 5.

3. Results

3.1. Effects of Maternal Exposure to RCO on Haematological Indices

There were significant increases in PCV, Hb and RBC in GD 14-21 treated rats as compared to control, while MCV and MCH decreased significantly in GD 14-21 as compared to control (Table 1). As also shown in Table 1, pregnant rats treated at gestation days 1-7 showed a reduction in packed cell volume when compared to control, while pregnant rats treated on gestation days 7-14, 14-21 and 1-21 showed an increase in PCV. These variations in values were however not significantly different from the control.

Table 1. Effects of RCO (950 mg/kg B.W.) administered to pregnant female rats at different gestation period on PCV, HB, RBC, MCV, MCH, and MCHC

3.2. Effects of Maternal Exposure to RCO on Percentage Leucocyte Cell Values

Percentage variations/changes in leucocyte cells values were all not statistically different from control values (Table 2).

Table 2. Effects of RCO (950 mg/kg B.W.) administered to pregnant female rats at different gestation period on leucocyte values

4. Discussion

An earlier study [14] (NTP, 1992) reported that there were no haematological abnormality except for statistically significant decrease in reticulocyte count after exposure of female rats to RCO. However, the study design was different from this study because 10 % RCO was added to diet and fed non pregnant rats for 13 weeks in the study. In this study, RCO was given orally to pregnant rats at different gestation periods.

There are several implications for marked variations in haematological parameters particularly during pregnancy. PCV may be altered and increased falsely as a result of dehydration while its decrease might indicate the presence of anemia as a result of blood loss or deficiency. Reduced MCV may be precipitated by iron deficiencies and thalasemia while increased MCV may follow Vitamins B12 and folate deficiencies. Pregnant rats given RCO at GD 14-21 were observed to show significant increase in PCV, Hb concentration and RBC when compared to control. The actual mechanism by which RCO influenced these is not known. However, enhanced erythropoiesis is known to be predicated on the duo of a highly functional endometrium at this stage of gestation and uteroferrin secreted by the endometrium. Efficient erythropoiesis is reported to require iron, folate, Vitamin B12 and erythropoietin [15]. Jepson, [16] also attributed increase in erythropoiesis late in pregnancy to erythropoietin transfer to the mother and that erythropoietic processes starts later in pregnancy and not early as fetal processes joins that of the mother. Furthermore, apart from erythropoietin, prolactin is known to increase as gestation days increase peaking at parturition [17]. Suggestion that prolactin may function as an erythropoietic stimulatory component in pregnant mouse has earlier been postulated by Jepsin and Lowenstein [18]. Jepsin and Lowenstein [19] further asserted that major increase in red blood volume occurs normally in late pregnancy as reticulocytes and plasma iron progressively decrease. It is therefore safe to suggest for now that RCO may have positively influenced any of these processes that have been identified to enhance erythropoiesis late at pregnancy.

On the other hand, the reduced erythropoiesis (though not statistically significant) observed in GD 1-7 treated rats may suggest that RCO use may be greatly contraindicated at early gestation period. In addition, the inhibiting effect on erythropoiesis induced by estrogen (levels of which are usually elevated at early gestation period) cannot be ruled out as it has been previously reported [17]. This usual inhibiting effect of estrogen can be further exacerbated by the estrogenic propensity of RCO which has been richly reported in literature [10,11,20,21]. Our previous study Salami and Raji, [10] gives credence to this line of thought by reporting higher estrogen levels in pregnant rats exposed to RCO at early gestation periods compared to late gestation period.

There were no significant differences in the differential white cells count of treated pregnant rats when compared to control. This suggests that RCO possibly has no overriding immuno-modulatory effects. Monocytes, Eosinophils and Basophils levels are known to increase during infection stress leukemia and inflammation while reduced neutrophils and lymphocytes are usually recorded during intake of some medications, autoimmune and bone marrow diseases and during severe infections [22].

Conclusively, RCO exposure at different gestation periods has no significant effect on differential leucocytes cells count. However, RCO exposure seems to militate and enhance erythropoiesis at early and late gestation period respectively.

References

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