本文關(guān)鍵詞：去甲澤拉木醛在大鼠腎移植模型中免疫抑制作用的研究　出處：《復(fù)旦大學(xué)》2013年碩士論文　論文類型：學(xué)位論文

  更多相關(guān)文章： 大鼠腎移植 急性排斥 去甲澤拉木醛 環(huán)孢素 雷公藤紅素

【摘要】：目的：為了更好地進(jìn)行移植免疫機(jī)理的探討、腎移植治療的研究和篩選新型的免疫抑制劑,探索建立更理想的大鼠原位腎移植模型。 材料與方法：用重250g、雄性SD和Wistar大鼠各10只作為腎移植的供、受體。暴露供體大鼠的左腎,在手術(shù)顯微鏡下解剖游離左腎動靜脈、輸尿管,阻斷左腎血管的上下腹主動脈,經(jīng)腹主動脈對左腎灌注0～4℃的腎保存液15～20ml,切取左腎、全長輸尿管帶部分膀胱瓣,修腎備用。切除受體左腎,用11—0顯微縫合線間斷端端吻合腎動脈,連續(xù)端端吻合腎靜脈,8—0線縫合帶膀胱瓣的輸尿管于受體膀胱。切除右腎。術(shù)后每日環(huán)孢素(Cyclosporin A, CSA)10mg/kg.d灌胃,大鼠移植術(shù)后7天,經(jīng)右眼內(nèi)眥靜脈抽血檢查腎功能并取存活鼠的移植腎作病理檢查。 結(jié)果：術(shù)后七天有8只大鼠存活,成功率80%,在術(shù)后第七天,大鼠的血肌酐、BUN正常范圍,與術(shù)前的血肌酐、BUN比較,P0.05,差別無顯著性。術(shù)后七天移植腎病理顯示腎組織少量淋巴細(xì)胞浸潤,腎小球、腎小管及微血管完好。 結(jié)論：在手術(shù)顯微鏡下建立大鼠左腎原位腎移植模型,能達(dá)到腎功能正常的要求,手術(shù)成功率較高,是可靠的大鼠移植模型。 目的：CSA是目前器官移植后最常用的免疫抑制劑之一,而雷公藤紅素是第一個(gè)從雷公藤中提取的單體。本試驗(yàn)以CSA和雷公藤紅素作為對照,觀察腎移植大鼠的生存期,觀察去甲澤拉木醛(T-96)對免疫抑制的實(shí)用價(jià)值。 材料與方法：以雄性SD、Wistar大鼠為供、受體,建立大鼠腎移植模型,依次進(jìn)入對照組和各實(shí)驗(yàn)組,每組8只,另外實(shí)驗(yàn)組每組多準(zhǔn)備2只,以備7天和14天時(shí)取移植腎送病理,不記入試驗(yàn)組。各組分別給予生理鹽水,CSA5mg/kg.d, CSA10mg/kg.d, T-9610mg/kg.d, T-9620mg/kg.d,雷公藤紅素0.3mg/kg.d和0.6mg/kg.d,每天灌胃一次。觀察記錄各組大鼠的生存天數(shù),取死亡鼠腎臟送病理檢查。 結(jié)果：去甲澤拉木醛能顯著延長大鼠的存活時(shí)間,且存在劑量效應(yīng)關(guān)系,低劑量和高劑量組生存時(shí)間分別為15.1±1.04d和21.4±2.61d,與對照組相比有顯著性差異(P0.01)。其存活時(shí)間與CSA組相當(dāng),差別無顯著性(P0.05)。雷公藤紅素低劑量組與T-96和CSA組相當(dāng),但高劑量生存時(shí)間較短(P0.05)。在7天腎臟病理檢查時(shí),對照組見顯著的排斥征象,而各給藥組不明顯。在14天移植腎病理檢查時(shí),對照組已無存活鼠,各低劑量組可見較為明顯的排斥,如大量淋巴細(xì)胞浸潤,腎小管內(nèi)大量蛋白尿,但腎小球仍較為完整。而T-9610mg/kg.d和CSA10mg/kg.d組僅可見少量淋巴細(xì)胞浸潤,腎小球和小管、微血管完好。 結(jié)論：本試驗(yàn)初步驗(yàn)證去甲澤拉木醛具有較強(qiáng)的免疫抑制作用,10mg/kg.d和20mg/kg.d均能顯著的延長大鼠移植腎的存活時(shí)間,且有劑量效應(yīng)關(guān)系,效果與CSA相仿。在高劑量,優(yōu)于雷公藤紅素。
[Abstract]:Objective: in order to better discuss the mechanism of transplantation immunization, we should research and screen new immunosuppressants for renal transplantation, and establish a more ideal orthotopic kidney transplantation model in rats.
Materials and methods: 250g male SD Wistar rats and 10 rats as renal transplantation for receptor. Exposure of left kidney donor rats, dissected the left renal artery and vein under surgical microscope ureteral occlusion of the left renal vessels, the abdominal aorta, abdominal aorta of left renal perfusion 0 to 4 DEG C kidney preservation solution 15 ~ 20ml, cut the left kidney and ureter with bladder flap, repair the renal reserve. The recipient's left kidney, with 11 - 0 suture line continuous end-to-end anastomosis of renal artery, continuous end-to-end anastomosis of renal vein, 8 - 0 line suture zone of the ureter to the bladder flap the recipient bladder. Resection of the right kidney. Postoperative daily cyclosporine (Cyclosporin A, CSA 10mg/kg.d) by gavage for 7 days, rats after transplantation, renal transplantation by right angular vein blood tests of renal function and survival of rats for pathological examination.
Results: seven days after the operation, 8 rats survived, the success rate of 80%, on the seventh day after operation, serum creatinine of rats, BUN in the normal range, and serum creatinine, preoperative BUN, P0.05, there was no significant difference. After seven days of transplant renal pathology of renal tissue showed that a small amount of lymphocyte infiltration. Glomerular, tubular and microvascular integrity.
Conclusion: a rat orthotopic renal transplantation model of left kidney can be established under operation microscope to achieve normal renal function and high success rate. It is a reliable rat transplantation model.
Objective: CSA is the most commonly used after organ transplantation and immunosuppression of tripterine is the first monomer extracted from Tripterygium wilfordii. In this experiment, CSA and tripterine as control rats, the survival of renal transplantation, to observe demethylzeylasteral (T-96) on immune suppression of practical value.
Materials and methods: male SD and Wistar rats as donors, receptor, rat kidney transplant model, in order to enter the control group and the experimental group, 8 rats in each group, the experiment group had to prepare more than 2 only, for 7 days and 14 days after transplantation of kidney pathology in experimental group. Each group respectively treated with normal saline, CSA5mg/kg.d, CSA10mg/kg.d, T-9610mg/kg.d, T-9620mg/kg.d, 0.3mg/kg.d and 0.6mg/kg.d of tripterine, orally once a day. The rats were recorded. The survival days of death from renal pathologic examination.
Results: demethylzeylasteral can significantly prolong the survival time of rats, and there was a dose effect relationship, low dose and high dose group survival time were 15.1 + 1.04d and 21.4 + 2.61d, compared with the control group there were significant differences (P0.01). The survival time and CSA group, no significant difference (P0.05). The celastrol and T-96 low dose group and CSA group, but high dose short survival time (P0.05). On the 7 day of renal biopsy, the control group saw significant signs of rejection, and each group is not obvious. On the 14 day of pathological examination, the control group has no survival in the low dose group showed obvious rejection, such as a large number of lymphocytic infiltration, renal tubular proteinuria, but still relatively complete. Glomerular T-9610mg/kg.d and CSA10mg/kg.d groups only showed a small lymphocytic infiltration, glomerular and tubular, microvascular intact.
Conclusion: this experiment demethylzeylasteral can inhibit the immune, 10mg/kg.d and 20mg/kg.d can significantly prolong the allograft survival time of rats, and the dose effect relationship, and the effect is similar to CSA. In high doses, better than tripterine.

【學(xué)位授予單位】：復(fù)旦大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2013
【分類號】：R699.2;R-332

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