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Please cite this paper as: Hadi V, Kheirouri S, Alizadeh M, Khabbazi AR, Hosseini H. Effects of Nigella sativa
oil extract on inflammatory cytokine response and oxidative stress status in patients with rheumatoid arthritis; a
randomized, double-blind, placebo-controlled clinical trial. Avicenna J Phytomed, 2016; 6 (1): 34-43.
each group to cover 30% dropout (Toubiet marital status), physical activity level, and
al., 2005).Patients were recruited from the psychological stress of the patients were
Sheykholrayis outpatient Clinic affiliated assessed using IPAQ and STAI-Y
to TBZMED from May 2012 to November questionnaires, respectively. Moreover,
2013. dietary intake of the participants was
The inclusion criteria consisted of (1) evaluated using a three-day dietary record
volunteer women patients aging between before and after the intervention. Dietary
20 to 50 with mild to moderate RA, data were analyzed using Nutritionist IV
according to 2010 ACR-EULAR criteria, software (First Databank, San Bruno, CA,
(2) being under treatment with USA).
methotrexate, hydroxychloroquine, and In order to establish compliance with
prednisolone less than 10 milligrams per treatment, all of the participants were
day (DMARDs), (3) not receiving any asked to keep unused capsules in drug
non-steroidal anti-inflammatory drugs or containers. All of the patients were
cytokine inhibitors with stable medication monitored by phone call every 15 days for
for at least 2 months prior to the any probable adverse events.
intervention, and (4) having body mass
index (BMI) less than 40. The exclusion Blood sampling
criteria of the study included those with (1) At the baseline and endpoint of the
pregnancy and lactating, (2) hormone study, after 10-12 hours fasting, venous
therapy or receiving oral contraceptives, blood was drawn and centrifuged for 15
(3) having any metabolic disorders such as minutes at 1500 g to obtain serum. Serum
diabetes mellitus, lactose intolerance, samples were stored at -20 ºC until
Cushing's syndrome, or thyroid biochemical analysis.
dysfunctions, (4) kidney or liver diseases,
(5) chronic inflammatory diseases Measurement of cytokines levels
including inflammatory bowel diseases, Serum levels of TNF-α and IL-10 were
and (6) any history of taking antioxidant or determined using commercially available
anti- inflammatory supplements 4 weeks cytokine ELISA kits (DIASource, Belgium
prior to the interventions or being on and eBioscience, USA, respectively)
weight reduction diets and smoking. following instructions of the manufacturers
Intervention group received two 500 at 450 nM wavelength in an ELISA plate
milligram capsules containing Nigella reader apparatus (Awareness, Statfax-2100
sativaoil each day (Ghetaet al., 2011) for 8 model, USA).
weeks. The other group consumed two
placebo capsules (paraffin) per day for the Measurement Antioxidant defense
same period of time (120 capsules per system (TAS, SOD, and catalase)
bottle, produced by Barij Asans of Kashan, Serum total antioxidant capacity (TAC)
Kashan, Iran). Nigella sativa oil and was determined using a Randox TAS kit
placebo were dispensed in capsules with (Randox Laboratory, UK) (Miller et al.,
identical appearance to ensure double 1993). Superoxide dismutase (SOD)
blindness of the study. Besides, Nigella activity was measured
sativa oil and placebo capsules were spectrophotometrically using a Ra nsod kit
encoded by another person blinded to the (Randox Laboratory, UK). Catalase
study. activity (CAT) was measured using the
At baseline and at the endpoint of the method proposed by Aebi (Aebiet al.,
study, weight and height were measured 1984). All tests were performed with an
and BMI was calculated. Moreover, automatic analyzer (Abbott model Alcyon
demographic information (such as age, 300, USA).
job, disease history, medication, and
Statistical analysis
The data were analyzed using SPSS
Results
version 16 (SPSS Inc., Chicago, IL, USA).
Subjects
Value of pless than 0.05 was considered
Fifty female patients with RA were
significant. The quantitative and
recruited in the present clinical trial and 42
qualitative data were presented as mean ±
women completed the study. Eight percent
SD and median (25th and 75th percentiles),
(8%) of the Nigella sativa group and 20%
respectively. The normality of variables
of placebo group did not complete the 8-
and homogeneity of variances were tested
week treatment course (Figure 1). No
using the Kolmogorov–Smirnov and
severe adverse effects of the treatment
Leven tests, respectively. Chi-squared test
were reported.
was used to compare the two groups for
background characteristics. Independent t-
Patients' characteristics
test or Mann–Whitney U tests were used to
Baseline characteristics of the patients
compare mean values between groups at
are presented in Table 1. Baseline
baseline. Mean values before and after the
characteristics of the patients did not differ
study period were compared within the
between the two group
groups using the paired t-test or Wilcoxon
Q2
Figure 1. Flow diagram of the progress through the phases of randomized trial of two groups.
† Independent Student t-test for age and BMI; Chi-squared test for menopausal status.
††Mann-Whitney U test.
* mean ± SD.
‡
Frequency (percent)
Table 2. Energy and macronutrient intake in the two experimental groups at baseline and throughout the
study.
At the end of the study, there was no scores calculated at the beginning of the
statistically significant difference in BMI intervention and physical activity level
value between or within the groups. There showed no significant difference between
were no differences between the two the two groups in baseline of the study.
groups at the beginning of study regarding The patients’ micronutrient intakes in
to doses and types of drugs. STAI-Y the two groups remained stable during the
study period. Analysis of dietary group. The DAS28 score was significantly
questionnaires, summarized in Table 2, lower in the Nigella sativa group as
indicated that there were no significant compared with the placebo group at the
differences in energy and macronutrient end of the study.
intakes at baseline and end of study course
in both of the groups. Biochemical data
Disease activity (DAS28) score was not As shown in Table 3, in the Nigella
significantly different between the two sativa group, serum IL-10 was
groups at baseline. At the end of the study, significantly increased at the end of the
DAS28 score significantly decreased in the study in comparison with the baseline
Nigella sativa group (p<0.05), but it did (p<0.01).
not change significantly in the placebo
Table 3. Effect of Nigella sativa as compared with placebo on inflammatory, antioxidant, and stress oxidative
biomarkers in female patients with rheumatoid arthritis.
TNF-α: Tumor necrosis factor-alpha; IL-10: Interlukin-10; SOD: Superoxide dismutase; TAC: Total antioxidant
capacity; MDA: Malondialdehyde; NO: Nitric oxaid. mean ± SD and median (percentiles 25 and 75) are presented
for normally and not normally distributed measures, respectively. Median of differences (percentiles 25 and 75) are
presented instead of mean difference (95% CI). †Mann-Whitney U test
** Based on ANCOVA adjusted for baseline measures and confounding factors (duration)
††† Wilcoxon test
* Independent t-test
*** Paired t-test
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