اثر اسانس آویشن شیرازی بر تولید یون سوپراکسید و نیتریک اکسید و بیان ژن‌های NADH اکسیداز و نیتریک اکسید سنتاز در سلول‌های ماکروفاژ

نویسندگان

مرکز بیوتکنولوژی دانشگاه شیراز، شیراز، ایران

چکیده

هدف: اثرات مهارکننده‌ی اسانس آویشن شیرازی بر تولید سوپراکسید و نیتریک اکسید و بیان ژن‌های NADH اکسیداز (NOX)  و نیتریک اکسید سنتاز (NOS) مورد بررسی قرار گرفت.
 
مواد و روش‌ها: اسانس گیاه توسط روش تقطیر جداسازی و اجزای تشکیل‌دهنده آن بوسیله کروماتوگرافی گازی متصل به طیف‌سنج جرمی مشخص شد. قدرت جاروب کنندگی اسانس بر روی رادیکال سوپراکسید و نیتریک اکسید به روش بیوشیمیایی مورد بررسی قرار گرفت. در مرحله بعد سلول‌های ماکروفاژ موشی تحریک‌شده با لیپوپلی ساکارید (LPS) با غلظت های مختلف اسانس تیمار شدند و بیان ژن‌های ان آ دی اچ اکسیداز و نیتریک اکسید سنتاز به روش Real Time PCR و همچنین میزان تولید یون سوپراکسید و نیتریک اکسید به روش‌های بیوشیمیایی مورد بررسی قرار گرفت.
 
نتایج: ترکیبات اصلی تشکیل‌دهنده اسانس گیاه آویشن، لینالوول (9/32 درصد)، کارواکرول (7/26 درصد)، تیمول (5/12 درصد)، پاراسیمن (5/4 درصد) و الفا- پینن (2 درصد) می‌باشند. اسانس آویشن در غلظت های پایین 70-100 میکروگرم/میلی‌لیتر قدرت جاروب کنندگی زیادی از آنیون‌های سوپراکسید (O2-) و نیتریک اکسید (NO) در مطالعات درون شیشه‌ای از خود نشان می‌دهد. اسانس در غلظت غیرسیتوتوکسیک، در ماکروفاژهای موشی تحریک‌شده توسط LPS باعث کاهش شدید تولید یون سوپراکسید و نیتریک اکسید شد. همچنین در سلولهای J774 (لاین ماکروفاژهای موشی) تحریک‌شده با LPS اسانس آویشن باعث کاهش بیان ام آر ان آ نیتریک اکسید سنتتاز و NADH اکسیداز شد.
نتیجه‌گیری: مشاهدات حاکی از آن است که فعالیت آنتی‌اکسیدانی اسانس آویشن احتمالا از طریق کاهش
تولید نیتریک اکسید، سوپراکسید و کاهش بیان ژن‌های نیتریک اکسید سنتتاز و NADH اکسیداز است.
 

کلیدواژه‌ها


عنوان مقاله [English]

The effect of zataria essential oil on superoxide and nitric oxide production and NADH oxidase and nitric oxide synthase in macrophage cells

نویسندگان [English]

  • Maryam Aminizadeh
  • Gholamreza Kavoosi
چکیده [English]

Objective: In this study, inhibitory effect of Zataria multiflora essential oil on superoxide and nitric oxide production and NADH oxidase (NOX) and nitric oxide synthase (NOS) expression was examined.





Materials and Methods: Zataria essential oil was obtained by hydro-distillation and chemical composition was analyzed by gas chromatography-mass spectrometry (GC-MS). Superoxide and nitric oxide scavenging effects of zataria essential oil were tested by biochemical assays. Then, LPS-stimulated macrophages were treated by different concentrations of essential oil and expression of superoxide-producing enzyme and nitric oxide-producing enzyme was investigated by Real-time PCR, also superoxide anion and nitric oxide levels was examined by biochemical assays.





Results: The main components of the zataria essential oil were linalool (32.9%), carvacrol (26.7%), thymol (12.5%), p-Cymene (4.5%) and α-Pinene (2%). Zataria essential oil sequestered superoxide anion (O2-) and nitric oxide (NO) in in vitro assay at low concentrations of 70-100 µg/ml. Zataria essential oil at non-cytotoxic level strongly reduced intracellular reactive oxygen species (ROS) and nitric oxide (NO) production in LPS-stimulated macrophages. Superoxide-producing enzyme and NO producing enzyme expression in LPS-stimulated murine macrophage (cell line J774) was declined by zataria essential oil.





Conclusion: Due to the results, the in vivo antioxidant activity of zataria essential oil may be attributed to down-production of NO and superoxide and also down-regulation of NOS and NOX expression.

کلیدواژه‌ها [English]

  • Nitric oxide
  • Superoxide anion
  • Nitric oxide synthase
  • NADH oxidase
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