اثر اپیوم بر مارکرهای استرس اکسیداتیو در رده سلولی hepg2

هدف: بررسی‌ها نشان می‌دهد که تریاک پس از تنباکو پرمصرف‌ترین ماده مخدر در بسیاری از کشورها، به‌ویژه ایران می باشد. قسمت اعظم مواد مخدر توسط کبد متابولیزه می شود. لذا، مصرف تریاک می تواند به‫طور مستقیم سبب آسیب سلول‫های کبدی شود. لذا در این مطالعه اثر تریاک بر سیستم آنتی اکسیدانتی بافت کبد بررسی شد.  مواد و روش‌ها: در این تحقیق از رده‌ سلولی سرطانی کبد انسان (hepg2) استفاده شد. در ابتدا زنده بودن سلول‫ها با استفاده از روش mtt مورد ارزیابی قرار گرفت. سپس ic50  تعیین شد و همان غلطت و یک غلظت پایین‫تر و یک غلظت بالاتر (در مجموع سه غلظت) برای مطالعات بعدی استفاده شد. سپس پراکسیداسیون لیپیدی، میزان اکسیدانتی تام، و آنتی اکسیدانت تام اندازه گرفته شد. فعالیت آنزیم‫های کاتالاز، گلوتاتیون پراکسیداز و سوپراکسید دیسموتاز با استفاده از کیت انجام شد. نتایج:  نتایج آنالیز آماری حاکی از اختلاف معنی‫داری در میزان مارکر های استرس اکسیداتیو بود (p< 0.001).  در گروه دریافت کننده اپیوم با دوز 60 و 70 میکرو گرم در میلی لیتر در مقایسه با گروه کنترل میزان tos و mda  افزایش معنی‫داری داشت (p< 0.001)، در حالی‫که میزان tac  کاهش یافت(p< 0.001).  همچنین فعالیت آنزیم های کاتالاز، گلوتاتیون پراکسیداز و سوپراکسید دیسموتاز در گروه‫های درمان شده با تریاک کاهش نشان داد (p< 0.001).  نتیجه‌گیری: نتایج این مطالعه نشان داد که تریاک می‫تواند سبب افزایش تولید رادیکال‫های آزاد در کبد شده و فعالیت آنزیم‫های آنتی اکسیدانتی را کاهش دهد.

effect of opium on oxidative stress markers in hepg2 cell line

aim: addiction is an important social and health problem in many middle eastern countries. studies show that opium is the most commonly used substance after tobacco in many countries, especially in iran. opium is obtained from the seeds of the papaver somniferum plant and contains more than 40 different alkaloids, the most important of which are morphine, codeine, papaverine, noscapine, and thebaine. various studies show that morphine, which is one of the most important substances in opium, increases the production of free radicals in the body and reduces the antioxidant capacity. opium consumption has various adverse health effects on the body. therefore, long-term use of this combination can be related to some pathological consequences, including neurological disorders, liver toxicity, kidney dysfunction, oxidative stress, and apoptosis. most drugs are metabolized by liver hepatocytes and excreted by kidney cells. therefore, opium consumption can directly cause damage to liver cells. free radicals play an important role in liver diseases. the increase of free radicals and on the other hand the decrease of antioxidants in the body caused liver toxicity. oxidative stress is caused by an imbalance between the production of free radicals and their neutralization inside the body by antioxidant defense mechanisms. oxidative stress is produced in various diseases, consumption of toxic substances, and old age, since it is not possible to investigate the effect of opium on human liver cells, and no study has been conducted in this field. the liver is the most important organ that is directly responsible for the metabolism and excretion of opium. therefore, in this study, the effects of opium on oxidative stress markers in hepg2 cell line were determined.material and methods: human liver cancer cell lines (hepg2) were used in this experimental study. at first, the cell line was placed in a 25 ml flask in the culture medium. dmem (dulbecco’s modified eagle’s medium) containing 10% fbs (fetal bovine serum), and 1% penicillin and streptomycin antibiotics were incubated in a 37°c incubator with 5% co2 . the cells were treated with different concentrations of opium (0-100ug/ml) for 24 hours. then ic50 was determined and then a lower concentration, ic50, and a higher concentration (three concentrations in total) were used for further studies.the lipid peroxidation was measured by thiobarbituric acid method. the total oxidant was measured using xylenol orange. the amount of total antioxidants was determined by the frap (ferric reducing / antioxidant power) method. the catalase, glutathione peroxidase, and superoxide dismutase enzyme activities were measured by colorimetry methods. the data was entered into spss software (version 20)  and analyzed using anova and tukey statistical tests. p less than 0.05 was considered a significant level. results: statistical analysis results indicated a significant difference in the amount of oxidative stress factors compared with control (p<0.001). in the group receiving opium with a dose of 60 and 70 mg/ml compared to the control group, the amount of tos and mda increased significantly (p<0.001), while the amount of tac decreased (p<0.001). also, the activity of catalase, glutathione peroxidase, and superoxide dismutase enzymes decreased in opium-treated groups (p<0.001). conclusion: the results of this research showed that opium had a lethal effect on hepg2 cells. also, opium caused an increase in oxidative stress in this cell line, which indicates the vulnerability of the liver against this compound. the results of this study show that although opium has analgesic effects, it can seriously damage the liver tissue.