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Antioxidant Activity and Cytotoxicity Effect of Various Extracts of Sclerorhachis Platyrachis on the Human Breast Adenocarcinoma Cells

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Antioxidant Activity and Cytotoxicity Effect of Various Extracts of Sclerorhachis Platyrachis on the Human Breast Adenocarcinoma Cells Scientific Journal of Kurdistan University of Medical Sciences No.104/ Mar-Apr 2020/33-42 Antioxidant activity and Cytotoxicity Effect of Various Extracts of Sclerorhachis platyrachis on the Human breast adenocarcinoma Cells Hasan Rezaei Seresht1, Hamid Cheshomi2, Leila Sadat Aldaghi3, Arezou Kaskani4 1. MSc of Phytochemistry, Traditional Complementary Medicine Reasearch Center, Sabzevar University of Medicinal Science, Sabzevar, Iran. (Corresponding Author), Tel: 051-44018319, E-mail: [email protected]. ORCID ID 0000- 0003-1303-5635 2. MSc, Cellular and Molecular Biology Reasearch Center, Sabzevar University of Medicinal Science, Sabzevar, Iran. ORCID ID: 0000-0001-7925-7797 3. MSc of Developmental Cell Biology, Cellular and Molecular Biology Reasearch Center, Sabzevar University of Medicinal Science, Sabzevar, Iran. 4. MSc of Pharmaceutical Chemistry, Department of Chemistry, Faculty of Science, Islamic Azad University of Sabzevar, Sabzevar, Iran. ABSTRACT Background and Aim: One of the spices of compositae is the Sclerorhachis platyrachis that is Iranian native. This aromatic plant has been known as bacterial growth inhibitor. This study have been attempted to investigate antioxidant activity and cytotoxicity effect of Sclerorhachis platyrachis extractson the MCF-7 cell line. Materials and Methods: The Hexane (HE), Chloroform (CE), Ethyl acetate (EE), Methanolic extracts (ME) of Sclerorhachis platyrachis leaves prepared by usingmaceration method. The DPPH assay was used for determination of antioxidant activity. Also, total phenolic content measured. The cytotoxicity of the extracts on MCF-7 and HEK-293 cells evaluated using MTT assay in both 48h and 72h. Results: evaluation of antioxidant activity indicated that radical scavenging ability was as ascorbic acid> ME > EE > CE > HE, While, total phenolic content was as CE>ME>EE>HE. The highest and the lowest cytotoxicity against MCF-7 cells were related to CE and ME respectively. Except for ethyl acetate extract, all extract were non toxic on the HEK-293 cells. Conclusion: Our findings showed that the chloroform extract of aerial part of Sclerorhachis platyrachis had the strongest effect against breast cancer carcinoma (MCF-7). Whereas, the extracts of this plant had no cytotoxicity on the normal cells. Keywords: Sclerorhachis platyrachis, Extract, HEK293 cell line, 1,1-DPPH Downloaded from sjku.muk.ac.ir at 19:29 +0330 on Wednesday September 29th 2021 Received: July 5, 2018 Accepted: Oct 21, 2019 How to cite the article: Hasan Rezaei Seresht, Hamid Cheshomi, Leila Sadat Aldaghi, Arezou Kaskani . Antioxidant activity and Cytotoxicity Effect of Various Extracts of Sclerorhachis platyrachis on the Human breast adenocarcinoma Cells. SJKU 2020; 25(1): 33-42 Copyright © 2018 the Author (s). Published by Kurdistan University of Medical Sciences. This is an open access article distributed under the terms of the Creative Commons Attribution-Non Commercial License 4.0 (CCBYNC), where it is permissible to download, share, remix, transform, and buildup the work provided it is properly cited. The work cannot be used commercially without permission from the journal مجله علمی دانشگاه علوم پزشکی کردستان / دوره بیست و پنجم / فروردین و اردیبهشت 99-24/9911 خاصیت آنتیاکسیدانی و اثر سایتوتوکسیک عصارههای مختلف گیاه مینای نیشابوری )Sclerorhachis platyrachis( بر روی سلولهای سرطان پستان حسن رضایی سرشت1، حمید چشمی2، لیﻻ سادات الداغی3 ، آرزو کسکنی4 1. کارشناسی ارشد فیتوشیمی، مرکز تحقیقات طب سنتی و جامع نگر، دانشگاه علوم پزشکی سبزوار، سبزوار، ایران )نویسنده مسئول(، تلفن: 99311044-41، پست الکترونیک: [email protected]، ، کد ارکید: 3333-3330-1030-4304 2. کارشناسی ارشد، مرکز تحقیقات سلولی مولکولی، دانشگاه علوم پزشکی سبزوار، سبزوار، ایران. کد ارکید: 3333-3331-7424-7747 0. کارشناسی ارشد زیست شناسی سلولی تکوینی، مرکز تحقیقات سلولی مولکولی، دانشگاه علوم پزشکی سبزوار، سبزوار، ایران. 9. کارشناسی ارشد شیمی دارویی، گروه شیمی، دانشکده علوم، دانشگاه آزاد اسﻻمی سبزوار، سبزوار، ایران. چکیده زمینه و هدف: گیاه مینای نیشابوری یکی از گونههای تیره کاسنی )Compositae( است که گونه بومی ایران است. این گیاه معطر به عنوان مهارکننده رشد باکتری معرفی شده است. در این مطالعه سعی شدهاست تا اثر آنتیاکسیدانی و سمیت سلولی عصاره این گیاه بر روی ردهسلولی MCF-7 و HEK-293 مورد ارزیابی شود. مواد و روشها: عصارههای هگزانی، کلروفرمی، اتیل استاتی و متانولی اندام هوایی گیاه مینای نیشابوری به روش خیساندن تهیه شد. بهمنظور تعیین خاصیت آنتی اکسیدانی از آزمون DPPH استفاده و همچنین محتوی فنولی کل اندازهگیری شد. سمیت سلولی عصارهها بر روی ردههای سلولی MCF-7 و HEK-293 در دو زمان 91 ساعت و 72 ساعت با استفاده از آزمون MTT مورد ارزیابی قرار گرفت. یافتهها: ارزیابی خاصیت آنتیاکسیدانی نشان داد که توانایی مهار به صورت اسکوربیک اسید<متانولی<اتیل استاتی<کلروفرمی<هگزانی بوده است. در حالی که محتوی فنولی کل بهصورت: کلروفرم<متانول<اتیل استات<هگزان بود و باﻻترین و کمترین سمیت سلولی بهترتیب مربوط به عصاره کلروفرمی و متانولی در مقابل ردهسلولی MCF-7 بود. تمامی عصاره- ها به جز عصاره اتیل استاتی برروی سلولی HEK-293 فاقد سمیت بودند. نتیجهگیری: - یافتههای ما حاکی از آن است که عصاره کلروفرمی اندام هوایی گیاه مینای نیشابوری، بهترین اثر را در برابر رده Downloaded from sjku.muk.ac.ir at 19:29 +0330 on Wednesday September 29th 2021 سلولی سرطان پستان )MCF-7( را دارا است. این در حالی است که عصارههای گیاه مورد نظر در برابر سلولهای نرمال-HEK 293 سمیتی ندارد. کلمات کلیدی: Sclerorhachis platyrachis، عصاره، رده سلولی HEK293 و 1,1-DPPH وصول مقاله: 19/11/47 اصﻻحیه نهایی: 27/7/41 پذیرش: 41/7/24 حسن رضایی سرشت 93 مقدمه به انحصاریبودن و همچنین غنیبودن این گیاه از متابولیت- آنتی اکسیدانتها با فرآیندهای فیزیولوژیکی و پاتولوژیکی های ثانویه، در این مطالعه بنیادی اثر عصارههای هگزانی، که مخصوصاً در آن رادیکالهای آزاد و گونههای فعال کلروفرمی، اتیل استاتی و متانولی برگ و ساقه گیاه مینای تولید میشوند مرتبطاند. تولید بیش از حد این گونهها در نیشابوری Sclerorhachis platyrachis بر روی رده- شرایط بیماری باعث ایجاد استرس اکسیداتیو در سلول سلولی MCF-7 و HEK293 مورد بررسی قرار گرفت. میشود و این در حالی است که سیستم آنتیاکسیدانی درون در تحقیقی که بر روی مواد معطر گیاه صورت گرفته، ROS سلول قادر به مقابله با اثرات مخرب همانند آنیون مشخص شده است که حاوی ترکیبات آلفا پینن، کامفر و بتا H O O - سوپر اکسید ) 2 (، پراکسید هیدروژن )2 2 (، رادیکال پینن است)13(. در تحقیقی دیگر که با هدف ارزیابی . 1O OH هیدروکسیل ) ( و اکسیژن یکتایی )2 ( نیست)1(. فیتوشیمایی گونههای خراسان صورت گرفت پس از آنالیز استرس اکسیداتیو شامل آسیب به مولکولهای زیستی نظیر وجود آلکالوئید و فﻻونوئید و نیز ساپونین وتانن تائید شده DNA ، لیپیدهای غشایی، آنزیمها است که در نهایت منجر است)17(. مشخص شده است که گیاه Sclerorhachis به بروز بیماریهایی نظیر سرطان و بیماریهای عصبی وابسته platyrachis دارای ترکیبات سسکوئی ترپنی و به سن میگردد)0-2(. فﻻونوئیدی میباشد)11(. همچنین در پژوهشی دیگر تعیین اثرات سودمند ترکیبات پلیفنولی برروی سﻻمت انسان در شده است که Sclerorhachis platyrachis فاقد درجه اول از طریق کاهش استرس اکسیداتیو بیان میشود. سمیت حاد برروی موش رت است)14(. باتوجه به وجود برخی از پلیفنولهای قادر به اعمال اثر ضدآپوپتوتیک، ضد مواد مؤثره جدید و نیز عدم سمیت این گیاه، این پژوهش با پیری، عﻻوه بر آن اثر ضد سرطان زایی و یا به طور کلی هدف ردیابی مواد مؤثره و همچنین تعیین میزان اثر مهار فرآیندهای تکثیر سلولی میباشند)4-9(. سیتوتوکسیسیتی و آنتی اکسیدانی هر عصاره صورت در سال 2312 سرطان پستان بهعنوان یکی از علل اصلی گرفت. مرگومیر ناشی از سرطان در زنان در سرتاسر جهان، با 1/7 میلیون مورد و نیز در حدود نیممیلیون مرگومیر شناخته مواد و روشها شده است)3(. این نوع از سرطان با تغییر تدریجی سلولهای گیاه مینای نیشابوری از کوه طبس شهر سبزوار جمعآوری Downloaded from sjku.muk.ac.ir at 19:29 +0330 on Wednesday September 29th 2021 نرمال به سلولهای سرطانی رخ میدهد. استروژن نقش گردید و عصارهگیری شدند. بهمنظور انجام آزمون سمیت اصلی را در رشد سلولهای اپی تلیوم پستان و در نتیجه سلولی، ردههای سلولیMCF-7 و HEK293 از بانک سرطان پستان بازی میکند)7(. سلولی انستیتو پاستور ایران خریداری شد. حﻻلهای مورد جنس Sclerorhachis متعلق به خانواده کاسنی چهار استفاده در این مطالعه از شرکت مرک خریداری گردید. گونه بوده که پراکندگی آنها محدود به ایران و افغانستان معرفهای DPPH و نیز استاندارد های نظیر گالیک اسید و است)1(. در فلور ایران دو گونه از آن آورده شده است که اسکوربیک اسید از شرکت سیگما خریداری شد. عمدتاً در شمال شرق کشور یافت میشوند)13-4(. جمعآوری گیاه Sclerorhachis platyrachis یکی از دو گونه گیاه مینای نیشابوری Sclerorhachis platyrachis در انحصاری ایران است که در خراسان رضوی گسترده شده اردیبهشتماه از کوه طبس در شمال شهر سبزوار جمعآوری است)19-11(. این گونه که روستاهای نزدیک سبزوار یافت شد. گیاه بدون آسیب خوردگی )به منظور حفظ اسانس میشود خواصی نظیر تسکین درد را دارا است)14(. با توجه گیاه( خشک گردید. سپس عصارهگیری توسط روش مجله علمی دانشگاه علوم پزشکی کردستان / دوره بیست و پنجم / فروردین و اردیبهشت 9911 93 خاصیت آنتی اکسیدانی... خیساندن انجام پذیرفت. به منظور عصارهگیری 03 گرم از گردید. در ادامه سلولها پس از شمارش در هر چاهک برگ گیاه مورد نظر، پودر شده و سپس به مدت 91 ساعت پلیت 43 خانه با غلظت 13333 سلول در هر سانتیمتر مربع سه مرتبه در مجاورت هر بار 43 میلیلیتر از حﻻلهای جای گرفتند )233 میکرولیتر در هر چاهک(. بهمنظور تعیین هگزان، کلروفرم، اتیل استات و متانول به ترتیب برای تهیه سمیت عصارهها، سلولها با غلظتهای بین 4 تا 133 عصارههای هگزانی، کلروفرمی، اتیل استاتی و متانولی )در µg/ml از عصارهها تیمار شدند. سپس 91 و 72 ساعت پس دمای اتاق( قرار گرفت. عصارههای بهدست آمده تحت از تیمار، میزان مرگومیر سلولها با استفاده از دستگاه الیزا شرایط خﻻء در دستگاه روتاری تغلیظ شد. در نهایت، ریدر از طریق سنجش مقدار بلور فورمازان تشکیل شده عصاره خشک بهدست آمده تا زمان آغاز آزمایشهای مورد ارزیابی قرار گرفت.
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