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. 2014 Dec;17(12):1012-9.

Thymoquinone, the main constituent of Nigella sativa, affects adenosine receptors in asthmatic guinea pigs

Laleh Pejman  1 , Hasan Omrani  1 , Zahra Mirzamohammadi  1 , Rana Keyhanmanesh  2
Affiliations

Thymoquinone, the main constituent of Nigella sativa, affects adenosine receptors in asthmatic guinea pigs

Laleh Pejman et al. Iran J Basic Med Sci. 2014 Dec.

Abstract

Objectives: For determining the mechanism of anti-asthmatic effect of thymoquinone, this investigation evaluated the effect of thymoquinone in the presence of selective A2A and A2B adenosine receptor antagonists (ZM241385 and MRS1706, respectively).

Materials and methods: Seventy guinea pigs were randomly divided to 7 groups; control (C), sensitized with ovalbumin (S), sensitized groups pretreated with thymoquinone (S+TQ), ZM241385 (S+Anta A2A), MRS1706 (S+Anta A2B), thymoquinone and antagonists (S+Anta A2A+TQ and S+Anta A2B+TQ). Thymoquinone and each of these antagonists with 3 mg/kg dose were injected i.p. on 10(th) day of sensitization protocol. Tracheal responsiveness (TR) to methacholine and ovalbumin (OA), and total and differential cell count in lung lavage fluid (LLF) in different groups were measured.

Results: Increased EC50 and LLF neutrophil count and decreased TR to methacholine and OA, LLF eosinophil and basophil counts were observed in S+TQ group compared to S group (P<0.001 to P<0.05). Significant decrease in EC50 (P<0.01), LLF neutrophil, lymphocyte and monocyte count (P<0.001 for all) and significant increase in TR to OA (P<0.01), LLF total WBC (P<0.01) and eosinophil count (P<0.001) were observed in S+A2A group compared to S+TQ group. There was significant increase in LLF eosinophil and monocyte counts in S+Anta A2B group compared with S+TQ group (P<0.001 for both). In S+TQ+Anta A2A group, there was significant increase in LLF eosinophil (P<0.001) and significant decrease in LLF neutrophil (P<0.01) and monocyte (P<0.001) counts compared with S+TQ group.

Conclusion: Thymoquinone affects adenosine receptors, which suggest that some of its anti-inflammatory effects may be mediated by these receptors.

Keywords: Adenosine receptor; Asthma; MRS1706; Thymoquinone; ZM241385.

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Figures

Figure 1
Figure 1
Cumulative log concentration-response curves of methacholine induced contraction of isolated trachea in control (C), sensitized (S), sensitized pretreated with thymoquinone (S+TQ), sensitized pretreated with selective A2A antagonist (ZM241385) and selective A2B antagonist (MRS1706) (S+Anta A2A and S+Anta A2B), sensitized groups pretreated with selective A2A antagonist and thymoquinone (S+Anta A2A+TQ) and selective A2B antagonist and thymoquinone (S+Anta A2B+TQ) guinea pig tracheal chains in the organ bath (for each group, n=7)
Figure 2
Figure 2
Individual values and mean±SEM (big symbols with bars) of tracheal response to methacholine (EC50) in control (C), sensitized (S), sensitized pretreated with thymoquinone (S+TQ), sensitized pretreated with selective A2A antagonist (ZM241385) and selective A2B antagonist (MRS1706) (S+Anta A2A and S+Anta A2B), sensitized groups pretreated with selective A2A antagonist and thymoquinone (S+Anta A2A+TQ) and selective A2B antagonist and thymoquinone (S+Anta A2B+TQ) (for each group, n=7). Statistical differences between control and different groups: ns: non significant differences, +; P<0.05, ++; P<0.01, +++; P<0.001. Statistical differences between pretreated groups vs sensitized group: NS: non significant differences, *: P<0.05
Figure 3
Figure 3
Individual values and mean±SEM (big symbols with bars) of tracheal contractility response to 10 μM methacholine in control (C), sensitized (S), sensitized pretreated with thymoquinone (S+TQ), sensitized pretreated with selective A2A antagonist (ZM241385) and selective A2B antagonist (MRS1706) (S+Anta A2A and S+Anta A2B), sensitized groups pretreated with selective A2A antagonist and thymoquinone (S+Anta A2A+TQ) and selective A2B antagonist and thymoquinone (S+Anta A2B+TQ) (for each group, n=7). Statistical differences between control and different groups: +; P<0.05, ++; P<0.01, +++; P<0.001. Statistical differences between pretreated groups vs sensitized group: NS: non significant differences, **; P<0.01
Figure 4
Figure 4
Individual values and mean±SEM (big symbols with bars) of tracheal response to ovalbumin (percent concentration in proportion to contraction obtained by 10 µM methacholine) in control (C), sensitized (S), sensitized pretreated with thymoquinone (S+TQ), sensitized pretreated with selective A2A antagonist (ZM241385) and selective A2B antagonist (MRS1706) (S+Anta A2A and S+Anta A2B), sensitized groups pretreated with selective A2A antagonist and thymoquinone (S+Anta A2A+TQ) and selective A2B antagonist and thymoquinone (S+Anta A2B+TQ) guinea pigs (for each group, n=7). Statistical differences between control and different groups: +; P<0.05, ++; P<0.01, +++; P<0.001. Statistical differences between pretreated groups vs sensitized group: NS: non significant differences, **: P<0.01
Figure 5
Figure 5
Individual values and mean±SEM (big symbols with bars) of total LLF WBC number in control (C), sensitized (S), sensitized pretreated with thymoquinone (S+TQ), sensitized pretreated with selective A2A antagonist (ZM241385) and selective A2B antagonist (MRS1706) (S+Anta A2A and S+Anta A2B), sensitized groups pretreated with selective A2A antagonist and thymoquinone (S+Anta A2A+TQ) and selective A2B antagonist and thymoquinone (S+Anta A2B+TQ) guinea pigs (for each group, n=6). Statistical differences between control and different groups: ++; P<0.01, +++; P<0.001. Statistical differences between pretreated groups vs sensitized group: NS: non significant differences, *; P<0.05
Figure 6
Figure 6
The percentages of eosinophil (a), neutrophil (b), lymphocyte (c), monocyte (d) and basophil (e) of lung lavage fluid in control, sensitized (S), sensitized pretreated with thymoquinone (S+TQ), sensitized pretreated with selective A2A antagonist (ZM241385) and selective A2B antagonist (MRS1706) (S+Anta A2A and S+Anta A2B), sensitized groups pretreated with selective A2A antagonist and thymoquinone (S+Anta A2A+TQ) and selective A2B antagonist and thymoquinone (S+Anta A2B+TQ) guinea pigs (for each group, n=15). Statistical differences between control and different groups: ns; non significant differences, +; P<0.05, ++; P<0.01, +++; P<0.001. Statistical differences between pretreated groups vs sensitized group: NS; non significant differences,*: P<0.05, **; P<0.01, ***: P<0.001
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