Journal of Medicinal Plants Research Vol. 5(31), pp. 6857-6863, 23 December, 2011 Available online at http://www.academicjournals.

org/JMPR ISSN 1996-0875 2011 Academic Journals DOI: 10.5897/JMPR11.1357

Full Length Research Paper

Effect of Nigella sativa and bee honey on pulmonary, hepatic and renal function in Sudanese in Khartoum state
Nahid Mahmoud AL Ameen1*, Faisal Altubaigy2, Tamanna Jahangir3, Idriss Abdalla Mahday4, Esmaeel Abdurrahman Mohammed5 and Omer Abdel Aziz Musa 6
1Department

of Basic Science, College of Dentistry, Jazan University, Jazan, Saudi Arabia. 2 College of Dentistry Jazan University, Jazan, Saudi Arabia. 3 Department of Toxicology, College of Pharmacy, Jazan University Jazan, Saudi Arabia. 4 National Ribatt University Teaching Hospital Clinical Chemistry Lab, Khartoum Sudan. 5 Department of Pathology, Faculty of Medicine Ribatt, University Khartoum, Sudan. 6 Department of Physiology, Faculty of Medicine, Ribatt University, Khartoum, Sudan.
Accepted 8 November, 2011

Nigella sativa seeds and Bee honey (BH) had been widely used in traditional medicine. In the present study we assessed their effect in treating asthma. A dose of 2 g of whole N. sativa seeds and 1 teaspoon full of BH per day were taken by the subjects -5 asthmatics and 22 non-asthmatics, 8 to 40 years in Khartoum for three month. They were also asked to smell crushed seeds that were warped in porous cloth three times a day. We assessed the effects of both on pulmonary, renal and hepatic function. Regarding pulmonary function, there was a significant increase in forced vital capacity (FVC) (P=0.023) in asthmatics' group and peak expiratory flow rate (PEFR) (P = was 0.049) in non-asthmatics' group. For evaluating the toxicity of both renal and hepatic functions, both were assessed before and after treatment. Hepatic enzymes were spared whereas serum creatinine in non-asthmatics and blood urea in asthmatics and in those having upper limit blood urea decreased significantly (creatinine (P=0.049) and urea (P value = 0.023, 0.017, respectively). In conclusion both N. sativa and BH seem to have some benefits to asthmatics with no hepato-renal toxicity. Key words: Nigella sativa, bee honey, hepato-renal toxicity, controls, asthmatics, non-asthmatics, pulmonary function. INTRODUCTION Nigella sativa and bee honey have been used all over the world to treat many different diseases since ancient time (Abedaziz and Kandeel, 2011; Tajk et al., 2008). BH has many medicinal properties like antibacterial activity (Olaitan et al., 2007), wound healing capacity (Cooper, 2008), anti-tumour (Fakoor and Pipelzadeh, 2007) and anti-oxidant properties (egarac et al., 2009). N. sativa is an annual herbaceous plant that belongs to the family (Ranunculaceae) (Abedaziz and Kandeel, 2011; Tajk et al., 2008). It contains carbohydrates, proteins, (AbdElAleem and El-Deeb, 2006), linoleic, oleic and palmitic acid (Atta, 2003) and high concentrations of thymoquinone (TQ) and thymol (Al-Saleh et al., 2006). It has many medicinal properties like; anti-oxidant (Meral et

*Corresponding author. E-mail: nahi93@maktoob.com. Tel: 00966549299713. Abbreviations: BH, Bee honey; FVC, forced vital capacity; PEFR, peak expiratory flow rate; TQ, thymoquinone; ROS, reactive oxygen species; NF-kB, nuclear factor-kappaB; ALP, alkaline phosphatase; AST, aspartate aminotransferase; ALT, alanine aminotransferase; FEV1, forced expiratory volume 1.

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al., 2001), anti-inflammatory (Hajhashemi et al., 2004) analgesic and other properties (Kaleem et al., 2006; Bhatti et al., 2009; Abd El-Aziz et al., 2005). Asthma is a chronic inflammatory disease of the airways, involving recurrent episodes of reversible airway obstruction, nonspecific airway hyper-responsiveness, (Hamelmann and Gelfand, 2001) and infiltration of many inflammatory cells, including eosinophils, mast cells, macrophages and neutrophils (Kelly et al., 1988). N. sativa is a potent bronchodilator (Keyhanmanesh et al., 2007). It reduces tracheal responsiveness (Keyhanmanesh et al., 2010) and inflammatory cells (Boskabady et al., 2011; Keyhanmanesh et al., 2010). BH relieves nocturnal cough (Paul et al., 2007). Kidney disorder, hypertention and diabetes are associated with vascular functional and structural changes (Paravicini and Touyz, 2006) that are due to generation of reactive oxygen species (ROS) (Rahman, 2007) and activation of the redox-sensitive transcription factor (NF-kB) (Morcos et al., 2002). TQ inhibits NF-kB (Sayed and Morcos, 2007). It may be a clinically valuable agent in the prevention of renal disease (Sayed-Ahmed and Nagi, 2007) N. sativa has hepatoprotective effect. It protects liver from many toxic metals (Kanter et al., 2005).
MATERIALS AND METHODS This study was performed in National Ribatt University in Khartoum in Sudan from November 2008 to February 2009 for partial fulfillment for the degree of Master of Science in Human Physiology. The investigations were carried out in Clinical Chemistry Laboratory of National Ribatt University Teaching Hospital. We carried out the study using both N. sativa and BH and doses were selected base on preliminary studies their data is not shown (EL-Kholy et al., 2009).

Study design A treatment regimen study was performed. The protocol was approved by the Ethics Committee of the Faculty of Medicine, National Ribat University. An informed consent and a questionnaire were given to all subjects. The questionnaire covered personal data, clinical examination and laboratory investigations. The subjects were asked to take one tea spoonful of natural BH and swallow 2 g of N. sativa after chewing it every night for three months. They were also asked to smell crushed seeds warped in porous cloth taking as deep inspiration as possible three times a day. The subjects were examined and investigated for pulmonary, hepatic and renal function at day zero and after three months. Prior to investigation, the procedure was explained to all subjects then 5 ml (5cc) of blood was collected from anticubital vein into a heparin container. Plasma was obtained by centrifugation for biochemical analysis to determine concentration of creatinine, urea, ALP, AST and ALT. Inclusion criteria for (Group 1) were; both sexes, age between 8 to 40, Sudanese living in Khartoum state and suffering from moderate to severe asthma and exclusion criteria were; non Sudanese, age below 8 or above 40, living outside Khartoum nonasthmatics or suffering from mild asthma. For (Group 2), inclusion criteria were both sexes, age between 8 to 40, Sudanese living in Khartoum state non-asthmatics and exclusion criteria were; non Sudanese, age below 8 or above 40, living outside Khartoum asthmatic.

Biochemical analysis and pulmonary function test Stocks and working solutions were maintained at 0C in a refrigerator. Serum was obtained by high speed centrifugation. The hepatic enzymes, urea and creatinine were measured by enzymatic colorimetric methods on an full automatic analyzer (selectra X-L fullautomation). For evaluating pulmonary function, we used peak flow-meter to measure the PEFR to determine the maximum flow rate in a single forced expiration and spirometer to measure FVC and forced expiratory volume1 FEV1. Statistical analysis Data analysis was performed using SPSS program. Paired-Sample T Test procedure that compares the means of two variables for a single group was used. The significance of difference was taken for P- value 0.05.

Plant material N. sativa seeds were purchased from Jeddah local market in Saudi Arabia and BH from Northern Darfoor state local market in western Sudan then the doses were prepared for every subject.

RESULTS
Chemicals and reagents Urea and creatinine kits, alkaline phosphatase ALP, aspartate amino-transferase (AST) and alanine aminotransferase (ALT) reagents were obtained from Biosystem Company. Peak flow meter (Wright peak flow meter) and spirometer (Micro medical pocket spirometer) and mouth pieces were obtained from physiology laboratory in National Ribbat University.

The total number of subjects recruited was 27. 5 of them were asthmatics (1st group) and the others were nonasthmatics (2nd group). Asthmatics' group composed of 3 males and 2 females. The non-asthmatics' group was composed of 12 males (54.5%) and 10 females (45.5%). Pulmonary function showed significant increase in FVC in asthmatics and a significant increase in PEFR in nonasthmatics while the FEV1 did not change in both groups (Table 1; Figures 1,2,3,4 and 5). Effect of treatment on hepatorenal function The hepatic enzymes were spared in both groups. Serum creatinine decreased significantly in non-asthmatics

Human subjects 5 Asthmatics (3 males and 2 females who were regular patients at Dr. Omer Abdel Azizs clinic) and 22 controls (12 males and 10 females) were randomly selected from staff members of Ribat University and their children.

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Table 1. Increase in FVC in asthmatics (significant) and in non-asthmatics (insignificant).

FVC in asthmatics before treatment 2.04

FVC in asthmatics after treatment 2.31

FVC in non-asthmatics before treatment 2.8827

FVC in non-asthmatics after treatment 2.9695

Significant improvement in asthmatics (P value = 0.23) and insignificant increase in non-asthmatics (P value = 0.66).

2.5 2
1.5

FVC in asthmatic before treatment

FVC in asthmatics after treatment

FVC in non-asthmatics before treatment

FVC in non-asthmatics aftertreatment

1 0.5

Figure 1. Significant increase in FVC in asthmatics. Significantly improved in asthmatics (P=0.023).

460 410 360 310 260 210

160

PEFR in asthmatic before treatment

PEFR in asthmatics after treatment

PEFR in non-asthmatics before treatment

PEFR in non-asthmatics after treatment

110 60 10

Figure 2. PEFR increased significantly in non-asthmatics. Significant improved in non-asthmatics (P=0.049).

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140.00000 120.00000 100.00000 80.00000 60.00000 40.00000 20.00000 .00000

Figure 3. Insignificant change in hepatic enzymes in asthmatics and non-asthmatics.
ALP in asthmatics before treatment ALP in asthmatics after treatment ALP in non-asthmatics before treatment ALP in non-asthmatics after treatment AST in asthmatics before treatment AST in asthmatics after treatment AST in non-asthmatics before treatment AST in non-asthmatics after treatment ALT in asthmatics before treatment ALT in asthmatics after treatment ALT in non-asthmatics before treatment ALT in non-asthmatics after treatment

1 0.9 0.8 0.7 0.6 0.5 0.4 0.3 0.2 0.1 0

Serum creatinine in asthmatics before treatment Serum creatinine in asthmatics after treatment Serum creatinine in nonasthmatic before treatment Serum creatinine in nonasthmatic after treatment

Figure 4. Significant reduction in serum creatinine in nonasthmatics. Significantly reduced in the non -asthmatics (P=0.049) while its reduction in asthmatics was insignificant.

(Figure 4 and Table 2. Blood urea decreased significantly in asthmatics and in those having upper margin blood urea in the nonasthmatics' group (Figure 5).

DISCUSSION The present study was designed to investigate the efficacy of N. sativa (2 g/day for 3 month) and

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Table 2. Significant reduction in blood urea in asthmatics and in those having upper margin blood urea in non-asthmatics.

Blood urea in asthmatic before treatment 34

Blood urea in asthmatics after treatment 25

Blood urea in nonasthmatics before treatment 22.0455

Blood urea in nonasthmatics after treatment 19.9545

Upper margin blood urea in non-asthmatics before treatment 34.5

Upper margin blood urea in non-asthmatics after treatment 21.25

Significant reduction in blood urea in asthmatics (P value = 0.29) and in those having upper margin in non-asthmatics (P value = 0.017).

35

30

blood urea in asthmatic before treatment

25

blood urea in asthmatics after treatment blood urea in non-asthmatics before treatment blood urea in non-asthmatics after treatment

20

15

10

Upper margin blood urea nonUpper margin blood urea inin nonasthmatics before treatmen asthmatics before treatment Upper margin blood urea in nonupper margin blood urea in nonasthmatics after treatment asthmatics after treatmen

0
Figure 5. Significant reduction in blood urea in asthmatics and in subjects with upper margin blood urea in the nonasthmatics. (P value = 0.29) in asthmatics and (P value = 0.017) in non-asthmatics.

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BH (tea spoonful daily for 3 month) in treating asthma. Their effect on hepatorenal function was also assessed to confirm the low toxicity of them. Boskabady et al (2004) found that N. sativa has a bronchodilator activity (Boskabady and Sheiravi, 2002; Boskabady et al., 2004). It improves the pulmonary function in asthmatics (Boskabady et al., 2010). This agrees with our finding. Nevertheless in our study the improvement in FVC was in the asthmatics' group while the PEFR increased significantly in the non-asthmatics. This may be due to having a small group for the study. Khader et al. (2009) found that TQ causes glutathione depletion and liver damage (Khader et al., 2009), but this finding is contradicted by many researchers like EL-Kholy et al. (2009) Qusti and Mahmoud (2009) (Kanter et al., 2005; Cemek et al., 2006; Yildiz et al., 2008; Al-Ali et al., 2008; EL-Kholy et al., 2009; Qusti and Mahmoud, 2007). Al-Ali et al. (2008) found that LD50 of TQ after intraperitoneal injection and oral gavage are 10 to 15 times and 100 to 150 times greater than doses reported for its anti-inflammatory, anti-oxidant and anti-cancer effects (Al-Ali et al., 2008). EL-Kholy et al. (2009) found that each of N. sativa and BH has hepatoprotective effect which was greater when they are combined. Our study showed that oral administration of N. sativa with BH has no toxicity in the doses used. These results agree with previous data reporting that N. sativa has a wide margin of safety. TQ acts as an anion scavenger that neutralizes oxygen radicals. It may have a beneficial adjunctive effect in improving kidney function. Creatinine is commonly measured as an index of glomerular function (Treasure, 2003). The normal range of serum creatinine is 0.7 to 1.3 for men and 0.6 to 1.1 women (Passos et al., 2003). In our study there was a significant reduction in serum creatinine in the non-asthmatics. Blood urea decreased in asthmatics' group and in those having upper margin blood urea in the non-asthmatics' group. This may be due to the fact that the subjects of asthmatics' group have higher blood urea relative to the non-asthmatics. All these changes were within the normal range. CONCLUSIONS AND RECOMMENDATIONS This study showed that N. sativa in combination with BH improves pulmonary function in asthmatics. Further studies with larger groups should be carried out.
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