@2024 Afarand., IRAN

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ISSN: 2252-0805    The Horizon of Medical Sciences 2016;22(1):13-20

Background
… [1-4] As a medical herb, Nigella sativa has been used to treat some diseases. Main effective materials in the aqueous extract of Nigella sativa are Thymoquinone, Dithymoquinone, Thymohydroquinone, and Thymol. Seeds of Nigella sativa contain fat, vitamins, protein, minerals, and carbohydrates [5, 6]. … [7-12]
Previous Studies
There is no study about the effects of the extracts of Nigella sativa on the functional damage of kidney caused by doxorubicin.
Aim(s)
The aim of this study was to determine the effects of hydro-alcoholic extract of the seeds of Nigella sativa on doxorubicin-induced functional damage of kidney in the rats.
Research type
This is an experimental study.
Research society, place and time
Male Wistar rats weighing 220-300g from Animal Room of Medicine Faculty of Mashhad University of Medical Sciences (Iran) were studied in 2014.
Sampling method and number
32 rats were studied.
Used devices&materials
The rats were kept at standard conditions (22±2°C and 12-hour darkness/light cycle) in Animal Room. Hydro-alcoholic extract of Nigella sativa was prepared through maceration. Seeds of Nigella sativa were bought and identified by botanists of Ferdowsi University (Iran). The rats were randomly divided into 4 groups each containing 8 rats. In control group, the rats had an open access to food and water, and at day 7, they received normal saline via tail vain injection in volume equal to the needed volume for doxorubicin injection. In doxorubicin group, 5mg/kg doxorubicin injection was done as tail vain injection [13] at day 7. In Nigella sativa plus doxorubicin group, 200mg/kg hydro-alcoholic extract treatment was done for 6 days and then, doxorubicin injection was done at day 7. And after that, 28-day oral administration of doxorubicin was done daily. In Nigella sativa group and as the rats of third group, 200mg/kg hydro-alcoholic extract of Nigella sativa treatment was administrated without doxorubicin injection. In all groups, doxorubicin injection was doe as tail vain. One day before the day zero and in days 5, 9, 13, 20, 27, and 34 of the study, the rats entered into metabolic cages for 24 hours and 24-hour urine samples, as well as blood samples from eye cavernous sinus, were collected. Urea concentration in serum and urine was measured, using Calorimetric Method based on the kit manual (Betagen; Iran) using a photometer tool (Convergys 100; Germany) on wave-length 578nm. Creatinine Concentration of serum and urine was measured through JAFFE Method based on the kit manual (Betagen; Iran) using a photometer device on wave-length 505nm. Glucose concentration of serum and urine was measured through GOD-PAP Method based on the kit manual (Betagen; Iran) using a photometer device at wavelength 546nm. Glomerular filtration was assumed as creatinine clearance, and it was computed by “Plasma Creatinine (mg/dl)/Urine Creatinine (mg/dl)×Urine Output (ml/minute)” formula. Urea clearance was computed by “Plasma Urea (mg/dl)/Urine Urea (mg/dl)×Urine Output (ml/minute)” formula. Kidney index was computed based on the body weight of the animal at day 35 and their right kidney weight by “Body Weight (g)/Kidney Weight (g)” formula. Data was analyzed, using SPSS 16 software. To compare the variables after investigating their normalization, the differences with control and doxorubicin groups were studied using One-way ANOVA and Tukey Post-hoc tests. And the differences between different days in the group were studied using Paired T-test. … [14]
Findings by Text
There was a significant increase in the mean serum urea at day 10 in doxorubicin group compared to the mean values of days before doxorubicin injection in the same group. There was no significant difference in the mean serum urea at different days between other groups. There was an increase in urea clearance in doxorubicin group at day 28 than other days. However, there was no significant difference in the mean urea clearance at different days between other groups. There was a significant reduction in the mean serum glucose at days 21 and 28 in doxorubicin group compared to the mean values of days before doxorubicin injection in the same group. There was no significant difference in the mean serum glucose at different days between other groups (Table 1). There was a significant difference in the mean concentration of urine glucose at days 28 and 35 in doxorubicin group compared to the mean values of days before doxorubicin injection in the same group. There was a significant reduction in the mean concentration of urine glucose at day 14 in Nigella sativa group compared to the mean values of days before doxorubicin in the same group. There was a significant increase in the mean concentration of urine glucose at day 28 in doxorubicin group compared to control group. There was a significant reduction in the mean concentration of urine glucose at day 28 in Nigella sativa plus doxorubicin group compared to doxorubicin group (Diagram 1). There was a significant reduction in the mean GFR at days 21 and 35 in doxorubicin group compared to the control day in the same group. There was a significant reduction in the mean GFR at day 35 in doxorubicin group compared to control group. There was a significant increase in the mean GFR at days 21, 28, and 35 in Nigella sativa plus doxorubicin group compared to doxorubicin group (Diagram 2). There was a significant increase in the kidney percentage in doxorubicin group (50.2±7.5) compared to control group (34.6±3.9). Nevertheless, there were no significant increases in Nigella sativa (38.1±8.2) and Nigella sativa plus doxorubicin (52.7±5.7) groups.
Main comparison to the similar studies
There was a significant reduction in the mean GFR in doxorubicin group compared to control group. The result is consistent with other studies. Doxorubicin reduces GFR [15]. Doxorubicin resulted in nephrotoxicity, which is consistent with other studies. Some mechanisms are an increase in NAG, GAG, glucose, and fibronectin excretion from urine, a reduction in the rate of GFR, an increase in the level of lipid-peroxidation and hydrogen peroxidase, and a reduction in the level of antioxidant enzymes [16]. Twice intravenous injection of doxorubicin on a 14-day interval for 20 days leads to tubular necrosis and glomerular membrane rupture, as well as an increase in the malondialdehyde level and a reduction in the level of total thiol groups in kidney tissue [17]. 6mg/kg doxorubicin injection leads to glomerular membrane rupture and inflammation, an increase in malondialdehyde level, and a reduction in the activity of superoxide dismutase enzyme in the kidney tissue [18]. Doxorubicin administration leads to glomerular sclerosis and injuries and uniformity in the podocyte [19]. Intravenous injection of doxorubicin leads to rupture and an increase in the podocyte feet appendages due to a reduction in the expression of the nephrin and podocin podocyte protein [20]. Different studies on tissues have shown glomerular vacuolization, inflammation, destruction and loss of epithelial cells, dilation and hyaline casts in the renal tubules in doxorubicin group [21]. There was a significant reduction in the mean glucose concentration in Nigella sativa plus doxorubicin group compared to doxorubicin group. In addition, there was a significant increase in the mean GFR in Nigella sativa plus doxorubicin group compared to doxorubicin group. The results are consistent with other studies. 3-week oral administration of 50mg/kg thymoquinone leads to tubular and glomerular damage improvement in the renal tissue damaged by doxorubicin [22]. There was a significant increase in the kidney index in doxorubicin group compared to control group, which was due to an increase in kidney weight and approximate reduction of body weight. Twice intravenous injection of doxorubicin on a 14-day interval for 20 days leads to a significant reduction in the weight of the animals treated by doxorubicin [20]. … [23-25] Doxorubicin administration leads to a reduction in the weight of rats treated by doxorubicin [26]. Extract of Nigella sativa had a preventive effect on oxidant activity of doxorubicin in the renal tissue. 3-week oral administration of 50mg/kg thymoquinone leads to a reduction in oxidative stress, an increase in the level of antioxidant enzymes such as superoxide dismutase, and a reduction in the level of tissue inflammation via a reduction in the production of TNF-α and interleukin-6 in the renal tissue damaged by doxorubicin [22]. … [27, 28]
Suggestions
Other doses of Nigella sativa should be studied in different administration methods and time intervals.
Limitations
Production of the extract needed too much time.
Conclusions
Hydro-alcoholic extract of the seeds of Nigella sativa leads to a reduction in the renal function damage caused by doxorubicin in the rats and results in an improvement in GFR level and glycosuria reduction.
Acknowledgments
Research Deputy of Mashhad University of Medical Sciences is appreciated.
Conflict of interest
Non-declared
Ethical Permissions
Medical Ethics Committee of Mashhad University of Medical Sciences confirmed the study.
Funding Sources
The study was funded by Research Deputy of Mashhad University of Medical Sciences.

TABLES and CHARTS

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