فعالیت کمپلمان‌‌ها در کشت اولیه سلول‌‌های طحال و راس کلیه ماهی شانک عربی (acanthopagrus arabicus) در مواجهه با دگزامتازون

داروها آلاینده‌‌های نوظهوری هستند که به دلیل تخلیه‌‌ آنها به اکوسیستم‌‌های آبی از طریق فاضلاب در سراسر جهان، نگرانی‌‌های زیادی را به‌خود جلب کرده‌‌اند. با این‌حال، مطالعات محدودی در مورد اثرات این آلاینده‌‌ها بر حیوانات آبزی وجود دارد. دگزامتازون یک گلوکوکورتیکوئید مصنوعی قوی با اثرات ضد التهابی و سرکوب‌کننده سیستم ایمنی است. مطالعه حاضر به بررسی تاثیر دگزامتازون بر فعالیت اجزاء کمپلمان سلول‌های راس کلیه و طحال ماهی شانک عربی (acanthopagrus arabicus) در شرایط آزمایشگاهی پرداخت. برای این منظور، سلول‌‌های طحال و راس کلیه کشت شده در مواجهه با غلظت‌‌های مختلف دگزامتازون (غلظت‌های آزمایشی 3، 1-10×3، 2-10×3 و 3-10×3 میکروگرم بر میلی‌‌لیتر برای طحال و غلظت‌‌های 10 × 3، 3، 1-10×3 و 2-10×3 میکروگرم بر میلی‌‌لیتر برای راس کلیه) قرار گرفتند. پس از 12، 24 و 48 ساعت، اثرات سمی دگزامتازون بر اجزاء سیستم کمپلمان بررسی شد. نتایج نشان داد که حساسیت سلول‌‌های طحال و راس کلیه کشت شده به دگزامتازون به طور وابسته به دوز افزایش می‌‌یابد. دگزامتازون به طور معنی‌‌داری محتوای c3 سلول‌‌های کشت شده را کاهش داد درحالی‌که تغییر قابل‌توجهی در محتوای c4 و ach50 سلول‌‌های کشت شده ایجاد نکرد. همچنین تاثیر دگزامتازون بر سلول‌‌های راس کلیه بیش از طحال بود. بر اساس نتایج، جنبه سرکوب‌کننده سیستم ایمنی دگزامتازون عمدتاً به دلیل اثرات سرکوب‌کننده آن بر سطح جزء c3 کمپلمان‌‌هاست. توصیه می‌‌شود در آینده، تاثیرات آلاینده دارویی دگزامتازون بر آبزیان بیشتری از جمله ماهیان مدل نیز انجام شود.

activity of complements in the primary culture of spleen and head kidney cells of arabian yellow fin sea bream (acanthopagrus arabicus) exposed to dexamethasone

introduction emerging anthropogenic pollutants in wastewater treatment plant (wwtp) effluents have been shown to pose a particular risk to aquatics and potentially affect the aquatic environment from the molecular to the ecosystem level (santos et al., 2010). micropollutants are emerging pollutants that include pharmaceuticals and personal care products (ppcps). due to the widespread consumption of a wide range of micropollutants and their discharge into wastewater worldwide, their occurrence and fate in aquatic environments has become a research topic in recent decades (ryu et al., 2014). the common wastewater treatment process is not able to completely remove these compounds from the wastewater, and as a result, a significant amount of these compounds enters the aquatic water environment (rehman et al., 2015). dexamethasone is a potent synthetic glucocorticoid with anti-inflammatory and immunosuppressive effects that is used to treat many diseases including allergies, asthma, covid-19, rheumatic problems, and skin diseases (sanders et al., 2020). today, the increasing use of dexamethasone from sources such as hospitals is a global concern. high levels of this drug, which is a cortisone derivative, have been detected in wastewater (herrero et al., 2013). as one of the important components of the innate immune system in ectothermic vertebrates, including fishes, the complement system has been less studied. evaluation of serum complement activity is considered as a valuable tool to diagnose the health status of fish. complement in the non-specific immune response to a compound can have a direct effect such as killing the pathogen by lysing it (ellis, 1999).methodology fifteen mature arabian sea bream, a. arabicus were collected from the bahrakan port in abadan. fish were transported to the 300 l tanks. after seven days of adaptation, the fish were anesthetized using 2-phenoxy ethanol (0.35 ml/l). then, the fish were dissected under aseptic conditions and their spleens and kidney heads were separated (wen et al., 2008). the separated tissues were immediately washed three times with 100 ml antibiotic medium (leibovitz’s l-15 (l-15) with 400 iu/ml penicillin, 400 µg/ml streptomycin and 200 µg/ml amphotericin b) each time for 30 minutes. the spleen cell culture was performed according to huang et al. (2009). the head kidney cell culture followed the protocols described by ribera et al. (2020). dexamethasone cytotoxicity was assessed by mtt method according to momeni et al. (2010). cultivated spleen and head kidney cells were plated in 24-well microplates (105 cells / ml l-15 medium / well) and incubated at 28°c for 24h. the cell viability was assessed using trypan blue exclusion test. then, 200 μl of fresh l-15 culture medium with different experimental concentrations of dexamethasone was added to each well and microplates were then incubated at 28°c for 48 h. each treatment runs in five replicates. for this purpose, the cell suspensions were collected at 0, 12, 24 and 48 hours of experiment and transferred to microtubes for further analysis. the level of c3 and c4 was measured using immunoturbidimetry method by a double–antibody sandwich elisa kits. ach50 was measured by hemolytic method according to sunyer and tort (1995).