Razi Journal of Medical Sciences

fa تاثیر کورکومین پوشش شده با نانوذرات کیتوزان بر فراموشی ناشی از اتانول در موش سوری نژاد NMRI Effect of chitosan nanoparticle enclosed curcumin on ethanol induced amnesia in NMRI mice فیزیولوژی Physiology پژوهشي Research زمینه و هدف: تأثیر استفاده از داروهای گیاهی با خواص آنتی‌اکسیدانی بالا از جمله کورکومین و همچنین روش‌های نانوتکنولوژی جهت بهبود فراموشی در مدل آلزایمر مورد مطالعه قرار گرفته‌اند. در این مطالعه پتانسیل نانوذرات کیتوزان حاوی کورکومین در بهبود فراموشی ناشی از اتانول مورد بررسی قرار گرفته است. روش کار: در این مطالعه تجربی، 128 سر موش سوری نژاد NMRI به وزن 40-30 گرم در سه آزمایش استفاده شد. تزریق بصورت درون صفاقی صورت گرفت. آزمایش اول جهت تعیین دوز مناسب اتانول به منظور تخریب حافظه، آزمایش دوم به منظور اثبات عدم ایجاد فراموشی توسط کورکومین، آزمایش سوم، جهت بررسی تأثیر دوزهای مختلف کورکومین پوشش شده با نانوذرات کیتوزان بر فراموشی ایجاد شده توسط اتانول انجام شد. داده‌ها در نرم‌افزار SPSS وارد شدند و توسط آزمون ANOVA و تست تعقیبی Tukey تجزیه و تحلیل شدند. 05/0 >p سطح معنی‌داری اول در نظر گرفته شد. یافته‌ها: آزمایش اول نشان داد که دوزهای kg/mg 1، 75/0 5/0 اتانول به طور معنی‌داری (001/0 >p و 01/0 >p) باعث فراموشی می‌شوند. آزمایش دوم نشان داد که کورکومین پوشش شده با نانوذرات کیتوزان باعث ایجاد فراموشی نمی‌شوند. آزمایش سوم نشان داد که همه دوزهای کورکومین با اختلاف معنی داری (001/0 >p) باعث بازگشت حافظه تخریب شده می‌شوند. ولی تزریق نانوذرات کیتوزان و سالین باعث بهبود حافظه تخریب شده نمی‌شوند. نتیجه‌گیری: نانوذرات کیتوزان استفاده شده در این مطالعه می‌تواند باعث اثرگذاری دوزهای پایین کورکومین در بهبود حافظه گردد. با این حال، تحقیقات بیشتری در استفاده از این نانوذرات جهت انتقال خوراکی داروها مورد نیاز می‌باشد. <div dir="ltr"></div> Background & Aims: Amnesia is known as central feature and the most prominent symptoms of Alzheimer’s disease (AD). Extracellular deposition of beta amyloid peptide and intracellular accumulations of tau protein are contributed in histopathological features of AD. The effect of medicinal herbs with high antioxidant property such as curcumin and nanotechnology approaches has been studied in amnesia improvement of Alzheimer animal model. Recently, positive effects of curcumin on neurogenesis as well as neuronal differentiation by activating brain drived neurotrophic factors have been shown, so that curcumin is being tested in phase II clinical trials. However, some curcumin features including insolubility in water, short half-life, low bioavailability and chemical instability at physiological pH have caused auxiliary use of nanotechnology to improve its biological function. Blood Brain Barrier (BBB) is considered as an obstacle for brain drug delivery. To overcome, different nanocarrier drug delivery systems are being studied. Amongst, chitosan nanoparticle has unique properties that can meet expectations. Chitosan, a cationic natural polysaccharide, is extensively used in central nervous system scaffold preparations systems in the last decade. It has been applied in for loaded drugs transportation to targeted organs in several cases of CNS and other disorders. Several advantages are considered for chitosan including biodegradability, biocompatibility, flexibility in surface modification and ease of multiple preparation methods. At the molecular level, chitosan exhibits its anti-Alzheimer effect by preventing kinase phosphorylation, inhibiting proinflammatory cytokines and blocking nitric oxide sythase which make it an attractive for nanocarrier forming material. In this study, potential of curcumin included Chitosan nanoparticles has been evaluated in improvement of amnesia induced by ethanol.  Methods: Curcumin was extracted from 10 gr Curcuma longa rhizome dried powder. The extracted components was dried, dissolved in the mixture of water and alcohol to purification of curcuminoid using electrochemically technique. The purified curcuminoid was neutralized using diluted phosphoric acid and dried in the shadow side. The purified powder of curcuminoids was characterized by FTIR. The yield was quantified and qualified using UV-Vis and HPLC. In this experimental study, 128 NMRI mice weighing 30-40mg were used in three phases. Animals were acclimated for two weeks, kept at 23 ± 2°C on a 12:12-h light-dark cycle (lights on 08:00-20:00) with free access to food and water. The mice were randomly divided into 16 groups (8 mice at each group). All experiments were carried out during the light phase between 9:00 and 14:00.  Injections were done intraperitoneally. The inhibitory passive avoidance tests were done using shuttle box apparatus which is consisted of two compartments of the same size (20×20×30 cm2) connected to each other and separated by a guillotine door. Floor and walls of one compartment was white while the other compartment's walls were dark with stainless steel bars in the floor. A  stimulator provided intermittent electric shocks (50 Hz, 3s, 1.5 mA intensity) to the grid floor of the dark compartment. Passive avoidance behavior included two consecutive days, training in the first day and behavioral test in the day after. Training included two steps: habituation and acquisition.  Habituation: Each animal was gently placed in the white compartment; after 5s, the animal was allowed to enter the dark compartment. The latency in which the animal entered the dark chamber was recorded. Latency more than 100s excluded animal from the experiments. Acquisition: thirty minutes after habituation step, animal put in the white chamber. Following complete enter to the dark chamber, the guillotine door was closed and a foot shock (50 Hz, 1 mA and 3s) was immediately delivered to the grid floor of the dark room. Twenty second later, the animal was returned to its cage. Two minutes later, animal was put again in the white chamber, the guillotine door was opened and the latency time was recorded. No enter to the dark chamber during 120s represents successful acquisition of inhibitory avoidance response. Behavioral test step: On the test day (the day after training) each animal gently placed in the white chamber and the guillotine door was opened 10s later. The latency time was recorded as indicator of inhibitory avoidance. Entering to the dark chamber wasn't followed by electric foot shocks. This study was conducted in three phases.The first phase was performed to get optimal ethanol dose can induce amnesia, the second phase to prove non impairing effect of nanoparticle enclosed curcumin, and the third phase to survey the effect of different doses of nanoparticle enclosed curcumin on amnesia induced by ethanol. Data was recorded and analyzed in GraphPad Prism software using ANOVA and Tukey post hoc test. P<0.05 was considered as first significant level.     Results: The first phase result showed that doses 0.5, 0.75, 1 mg/kgBW of ethanol can significantly induce amnesia. The second phase result showed non impairing effect of nanoparticle enclosed curcumin on memory function. The third phase result showed that all doses of nanoparticle enclosed curcumin can significantly (p<0.001) improve amnesia, but chitosan nanoparticle alone and saline couldn't. Conclusion: Recently has been shown that cognitive functions, learning and memory are strongly affected by ethanol consumption. According to the results, ethanol can induce memory impairment dose-dependently. Besides, chitosan used in this study didn't impair memory function. The result that has been shown by such previous studies. Hence, chitosan has widely used as an appropriate nanocarrier in drug delivery  system. Chitosan nanoparticles used in this study made even lower doses of curcumin to be effective in memory improvement while it has been reported that higher but not lower doses of curcumin can improve impaired memory. Therefore, a range of curcumin doses from high to low was investigated in this study. Drug delivery to the brain tissue requires passing blood brain barrier without any damage to its structure which is one of the most challenges in brain drug delivery. One approach is using anoparticle such as chitosan. It can be suggestad that chitosan not only promote curcumin solubility and bioavailability but also help drugs to pass blood brain barrier. Since, curcumin loaded in chitosan was deliverd to in vivo via intraperitoneal injection, However, more studies are needed for the application of this nanoparticle in oral drug delivery. کورکومین, کیتوزان, نانوذرات, اتانول, فراموشی Curcumin, Chitosan, Nanoparticle, Ethanol, Amnesia 26 36 http://rjms.iums.ac.ir/browse.php?a_code=A-10-5087-1&slc_lang=fa&sid=1 Goljahan Iri گل جهان ایری goli_iri@yahoo.com 3900319475328460059015 3900319475328460059015 No Islamic Azad University, Gorgan Branch, Gorgan, Iran گروه زیست شناسی، گرگان، ایران Nasrinsadat Azami نسرین سادات اعظمی azaminasrin@gmail.com 3900319475328460059016 3900319475328460059016 Yes Islamic Azad University, Gorgan Branch, Gorgan, Iran گروه زیست شناسی، گرگان، ایران Mahmoud Heidari محمود حیدری mahmodh78@gmail.com 3900319475328460059018 3900319475328460059018 No Islamic Azad University, Gorgan Branch, Gorgan, Iran گروه زیست شناسی، گرگان، ایران mehdi Eabdi مهدی عبادی Mehdi_2222002@yahoo.com 3900319475328460059017 3900319475328460059017 No Islamic Azad University, Gorgan Branch, Gorgan, Iran واحد گرگان، گروه شیمی، گرگان، ایران