پایان نامه های دامپزشکی(1)

 

رابطه بین شاخص های استرس اکسیداتیو و پارامترهای خون شناسی و پروفایل آهن در گوساله های شیری نوزاد

نوع مادهپایان نامه

نویسنده/استاد راهنما: رجبیان، فاطمه;;حیدرپور، محمد;مهری، مهرداد

سال دفاع:‎۱۳۹۳

مطالعه ی حاضر با هدف بررسی روند تغییرات شاخص های استرس اکسیداتیو همزمان با تغییرات خون شناسی و پروفایل آهن و همچنین ارتباط بین آن ها در گوساله های شیری تازه متولد شده انجام شده است. به این منظور‎30 رأس گوساله نژاد هولشتاین به ظاهر سالم به تعداد مساوی از نظر جنسیت (نر و ماده بودن) انتخاب شدند.تمامی گوساله های مورد مطالعه طی زایمان طبیعی متولد شده و گوساله‌ها به گونه ای انتخاب شدند که از نظر شکم زایشی مادرانشان در سه گروه ده تایی یک شکم زایش، دو شکم زایش و سه شکم زایش قرار گرفتند. نمونه‌گیری از هر گوساله در ‎5 نوبت انجام شد. خون‌گیری در زمان 48-‎24 ساعت اول پس از تولد و پس از آن در روزهای ‎7 و ‎14 و ‎21 و ‎28 بعد از تولد صورت گرفت. نمونهصگیری از ورید وداج با استفاده از ونوجکت با سرسوزن شماره ‎18 انجام گرفت. از هر گوساله cc10خون گرفته شد که حدود cc2 از آن به لوله حاوی ماده‌ی ضد انعقاد EDTAجهت انجام آزمایشات خون شناسی و تهیه ی همولیزه ی گلبولهای قرمز خون و الباقی آن به لوله بدون هیچ گونه افزودنی، جهت لخته شدن و جدا شدن سرم، منتقل گشت. بر روی هر نمونه، آزمایشات خون‌شناسی و بیوشیمیایی شامل اندازه‌گیریMCHC،MCH،MCV، PCV،Hb،RBC،WBC،Plt، پروتئین تام، فیبرینوژن، آهن،TIBC و درصد اشباع ترانسفرین و همچنین بررسی شاخص‌های استرس اکسیداتیو شامل اندازه گیری مالون دی آلدهید همولیزه گلبول های قرمز و مالون دی آلدهید، گروه تیول وظرفیت تام آنتی‌اکسیدانی(FRAP) سرم خون انجام گرفت. پس از اندازه‌گیری پارامترها و انجام تحلیل‌های آماری،مقادیر هماتوکریت، هموگلوبین،MCV، MCHو گروه تیول (به عنوان شاخص آنتی اکسیدانی) یک روند نزولی را در طی ماه اول زندگی نشان دادند و کمترین مقدار آن ها در انتهای دوره دیده شد. برعکس، مالون دی آلدهیدهمولیزه و سرم خون روند صعودی را نشان دادند. پروفایل آهن شامل آهن سرم خون،TIBC و درصد اشباع ترانسفرین نیز از یک روند صعودی در طی مطالعه برخوردار بودند. از لحاظ FRAP تغییر معنی داری در طی زمان مشاهده نگردید. به دنبال ارزیابی ارتباط بین شاخص های مختلف با استفاده از آزمون پیرسون، رابطه مثبت معنی داری بین مقدار FRAP و غلظت آهن سرم خون و TIBC در ‎7 و ‎21 روزگی، رابطه منفی معنی داری بین مقدار مالون دی آلدهید گلبول قرمز و تعداد گلبول های قرمز در 48-‎24 ساعت ابتدایی پس از تولد و ارتباط مثبت معنی داری بین FRAP و مقدار PCV غلظت هموگلوبین و تعداد گلبول های قرمز در ‎21 روزگی نشان داده شد. همچنین به دنبال مقایسه استرس اکسیداتیو و پروفایل آهن در دو گروه کم خون و غیر کم خون، مقدار مالون دی آلدهید گلبول های قرمز در گروه کم خون افزایش معنی داری در مقایسه با گروه غیر کم خون در انتهای دوره نشان داد (0.05>p). آهن و TIBC نیز در گروه کم خون کاهش معنی داری در مقایسه با گروه غیر کم خون در زمان های مختلف نشان دادند (0.05>p). روند صعودی مالون دی آلدهید در گوساله های مورد مطالعه، همراه با افزایش مقدار مالون دی آلدهید گلبول های قرمز در گوساله‌های کم خون در مقایسه با گوساله های غیر کم خون و رابطه منفی معنی دار بین مقدار مالون دی آلدهید گلبول قرمز و تعداد گلبول های قرمز می توانند شواهدی از نقش استرس اکسیداتیو در آسیب گلبول های قرمز و بروز کم خونی در هفته های ابتدایی زندگی گوساله باشند. عدم تغییر معنی دار FRAP (به عنوان شاخصی از ظرفیت آنتی اکسیدان های غیر پروتئینی خون) و حتی روند کاهشی گروه تیول می توانند به عنوان شواهدی از عدم کفایت سیستم آنتی اکسیدانی بدندر بدو تولد در دفاع علیه تولید بیش از حد رادیکال های آزاد و فراهم شدن زمینه رخداد استرس اکسیداتیو در ابتدای زندگی گوساله در نظر گرفته شوند.

 

The relationship betweenoxidative stress markers and hematology parameters and iron profilesin neonatal dairy calves BY Fateme Rajabian

 

 

The aim of this study was to investigate the relationships between oxidative stress markers, hematology and iron profile in neonatal dairy calves. Thirty Holstein calves were used for the experiment. Blood samples were collected from all of the calves within 24–48 hours after birth and at 7, 14, 21 and 28 days of age. Anti-coagulated blood was analyzed shortly after collection for preparation of erythrocyte hemolysate and measurement of hematological parameters including number of red blood cell (RBC), hemoglobin (Hb), heamatocrit (HCT), total leukocyte count (WBC), Platelet (Plt), MCH, MCV, MCHC, total protein and fibrinogen concentration. The amounts of iron profile including serum iron, TIBC, transferrin saturation, and oxidative stress markers including malondialdehyde (MDA), total antioxidant capacity (FRAP) and thiol groups were measured in the serum samples. In addition, the concentration of MDA was measured in the erythrocyte hemolysate prepared from the anti-coagulated blood. 94 1 / 4 During the experiment, HCT, Hb, MCH, MCV and thiol groups showed a quite similar pattern of changes and decreased during the first weeks of life. The lowest values were observed at the end of the study. In contrast, MDA concentrations in serum and hemolysate samples and iron profile showed a increasing trend during the study. The evaluation of relationship of oxidative stress markers with hematological parameters and iron profile showed the following results: a significant positive correlation between FRAP and serum iron and TIBC at days 7 and 21 of age (P<0.05), a significant negative correlation between erythrocyte MDA and RBC at 24-48 hours after birth (P><0.05), and a significant positive correlation between FRAP and HCT, Hb and RBC at day 21 of age (P><0.05). In addition, the comparison of oxidative stress markers and iron profile in anemic and non-anemic calves presented increased erythrocyte MDA in the anemic calves at the end of study and decreased serum iron and TIBC in the anemic calves in different sampling times. Increasing trend of MDA concentration and significant negative correlation between erythrocyte MDA and RBC along with increased erythrocyte MDA in the anemic calves may link erythrocyte damage in neonatal dairy calves to oxidative stress. Decreasing concentrations of thiol groups and unchanged concentrations of FRAP (as markers of antioxidant capacity) could be indicatives of inability of body antioxidant system against free radical generation during the first weeks of life in dairy calves. Keywords: Malondialdehyde, Oxidative stress, Red blood cells, Iron profiles, Calves>In addition, the comparison of oxidative stress markers and iron profile in anemic and non-anemic calves presented increased erythrocyte MDA in the anemic calves at the end of study and decreased serum iron and TIBC in the anemic calves in different sampling times. Increasing trend of MDA concentration and significant negative correlation between erythrocyte MDA and RBC along with increased erythrocyte MDA in the anemic calves may link erythrocyte damage in neonatal dairy calves to oxidative stress. Decreasing concentrations of thiol groups and unchanged concentrations of FRAP (as markers of antioxidant capacity) could be indicatives of inability of body antioxidant system against free radical generation during the first weeks of life in dairy calves.

 

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بررسی تاثیر افزایش طول عمر سلول های عضلانی ترشح کننده آنتی ژن هماگلوتینین از ویروس آنفلوانزای پرندگان (H5N1) بر القای پاسخ های ایمنی همورال و سلولی

نوع مادهپایان نامه

نویسنده/استاد راهنما: طباطبائی‌زاده، الیاس;;دهقانی، حسام

سال دفاع:‎۱۳۹۳

A واکسن را می توان به عنوان روشی موثر جهت پیشگیری از عفونت آنفلوانزای پرندگان H5N1 مورد توجه قرار داد هر چند که سطوح پایین بیان آنتی ژن می تواند محدود کننده پاسخ های ایمنی ایجاد شده بوسیله DNA واکسن باشد. در این مطالعه، تجویز همزمان DNA کد کننده پروتئین آنتی آپوپتوتیک XIAP به عنوان فاکتور تنظیم کننده آپوپتوز و تحریک کننده مسیرهای سیگنالینگ التهابی به عنوان روشی جهت ارتقای پاسخ های ایمنی ایجاد شده بوسیله DNA واکسن بیان کننده پروتئین H5 آنفلوانزای فوق حاد پرندگان مورد بررسی قرار گرفته است. نتایج کشت سلول نشان داد که DNA واکسن کد کننده آنتی ژن H5 قادر است باعث ایجاد آپوپتوز شود و این فعالیت پرو آپوپتوتیک H5 به میزان قابل توجهی با بیان همزمان پروتئین کامل XIAP یا موتانت XIAP( RING) در مدت ‎24 ساعت کاهش پیدا کرد. با این حال، پروتئین کامل XIAP قدرت بیشتری را در مقایسه با XIAP( RING) در جلوگیری از آپوپتوز القا شده با H5 نشان داد. همچنین توانایی ایمن سازی فرم های داخل غشایی و ترشحی H5 فیوز به چهار کپی از پپتید P28 مورد بررسی قرار گرفت. موش هایی که علاوه بر DNA واکسن، پلاسمید کد کننده XIAP را نیز دریافت کردند، تیتر آنتی بادی در آن ها افزایش پیدا کرد. موش های واکسینه شده با فرم ترشحی H5، تیتر آنتی بادی HI بالاتری را در مقایسه با موش های واکسینه شده با فرم داخل غشایی H5 نشان دادند. علاوه بر این، تجویز همزمان XIAP همراه با فرم ترشحی H5 منجر به پاسخ های قویتر آنتی بادی در مقایسه با فرم وحشی داخل غشایی H5 گردید. نتایج بدست آمده از این مطالعه نشان می دهد که برای طراحی DNA واکسن ها بخصوص مواردی که آنتی ژن مورد نظر فعالیت پرو آپوپتوتیک دارد، استفاده از ادجوانت مولکولی آنتی آپوپتوتیک و فرم ترشحی آنتی ژن می تواند بر روی پاسخ های ایمنی تاثیرگزار باشد.

 

 

DNA vaccine represents a powerful approach for prevention of avian H5N1 influenza infection, although DNA vaccine induced immune responses might be limited by the low levels of antigen expression. As a strategy to enhance adaptive immune responses elicited by a highly pathogenic avian influenza hemagglutinin 5 (H5) DNA vaccine, we explored the effect of co-administration of a DNA encoding X-linked inhibitor of apoptosis protein (XIAP) as a modulator of apoptosis and a stimulator of inflammatory signaling. We found that the DNA vaccine encoded H5 antigen was a potent stimulator of apoptosis and the H5 proapoptotic activity was significantly suppressed by co-expression of full length XIAP or mutant XIAP, deleted really interesting new gene (∆RING) zinc finger domain, in cultured cells as early as 24 hours (h). However, full length XIAP showed a higher potency than XIAP(∆RING) in the inhibition of H5-induced apoptosis. We also compared the immunizing ability of transmembrane and secretory forms of H5 and found that mice co-administered twice (with 3 weeks interval) with a plasmid encoding XIAP showed increased antibody titers. Mice vaccinated with the secretory form of H5 showed higher hemagglutination inhibition (HI) antibody titers than mice vaccinated with the transmembrane form of H5. Furthermore, co-administration of XIAP with the secretory form of H5 resulted into a stronger antibody response than the transmembrane form of H5. Our findings suggest that in the design of DNA vaccines for a given pro-apoptotic antigen, using an anti-apoptotic molecular adjuvant and the secretory form of antigen may have effects on the immune responses. Key words: apoptosis; avian H5N1 influenza; DNA vaccine; XIAP

 

 

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