發(fā)布時間：2018-06-07 02:06

  本文選題：一氧化氮 + 紅細(xì)胞變形性��； 參考：《瀘州醫(yī)學(xué)院》2012年碩士論文

【摘要】：一氧化氮(NO)在生物體內(nèi)作為一種反應(yīng)性極強(qiáng)的自由基，具有廣泛的生理作用。NO通過結(jié)合紅細(xì)胞血紅蛋白(Hb)中保守的β~(93)Cys殘基(Hbβ~(93)Cys)形成S-亞硝基血紅蛋白（SNO-Hb）。而Hb分子內(nèi)NO能從血紅素轉(zhuǎn)移至Hbβ~(93)Cys巰基，調(diào)節(jié)NO的貯存與釋放，介導(dǎo)血管舒張。庫血紅細(xì)胞隨著保存時間的延長，NO會發(fā)生大量流失，輸入體內(nèi)后紅細(xì)胞的擴(kuò)血管活性、變形能力及血紅蛋白攜氧能力等生理活性下降。因此，了解紅細(xì)胞存儲過程中NO的流失問題，認(rèn)識NO在循環(huán)系統(tǒng)中的作用機(jī)理，對保障紅細(xì)胞輸注的安全性和有效性具有極其重要理論意義和臨床價值。本研究擬通過體外補(bǔ)充NO來改善庫存懸浮紅細(xì)胞的形態(tài)和功能，，探討NO對紅細(xì)胞形態(tài)學(xué)及儲存損傷的影響機(jī)制，為紅細(xì)胞的有效保存，提供可參考的理論依據(jù)。目的：本次實驗對保存期間的懸浮紅細(xì)胞補(bǔ)充不同濃度梯度的NO試劑：L-Arginine(L-精氨酸/NO前體），分析檢測懸浮紅細(xì)胞不同保存時相的紅細(xì)胞內(nèi)NO量的改變及紅細(xì)胞變形性等相關(guān)指標(biāo)，以期探討NO對庫血懸浮紅細(xì)胞變形性的影響機(jī)制，從而進(jìn)一步評價儲存懸浮紅細(xì)胞的儲存質(zhì)量。 方法：取10例各1u新鮮懸浮紅細(xì)胞，4℃保存，試驗前分別取出4mL于無菌試管，總計500管，進(jìn)行編組編號，保存于專用儲存冰箱內(nèi)。在儲存的不同時相（3h、1d、3d、5d、7d、14d、21d、28d），每例樣本各取5管，其中1管不做處理，為對照組（A組），另外4管分別和不同濃度L-精氨酸（NO前體）混合培養(yǎng)，濃度分別為1μM（B）、5Μm(C)、10Μm(D)、50Μm(E)。培養(yǎng)1小時后測定觀察指標(biāo)。主要有：（1）紅細(xì)胞變形性相關(guān)指標(biāo)：血液高切粘度、低切粘度、卡松粘度、血漿粘度、紅細(xì)胞變形指數(shù)、聚集指數(shù)、剛性指數(shù)：（2）NO熒光探針檢測：通過測定平均熒光強(qiáng)度（PKPosX）反應(yīng)紅細(xì)胞內(nèi)的一氧化氮水平。 結(jié)果：保存期內(nèi)對照組和實驗組紅細(xì)胞變形指數(shù)、聚集指數(shù)和剛性指數(shù)均隨著時間延長而逐漸增高，且變化顯著。對照組高切相對粘度在存放1d后出現(xiàn)升高，隨后維持相同數(shù)值，遞增趨勢不明顯；低切粘度在3h—5d期間無顯著差異，14d-21d期間持續(xù)升高；卡松粘度在（5d、7d、14d、21d）時間點呈逐漸升高的趨勢。當(dāng)NO前體濃度為5μM時能夠明顯改善保存14d內(nèi)的庫血紅細(xì)胞變形性指數(shù)和3d-7d內(nèi)的剛性指數(shù)；NO前體濃度為10μM時，能明顯改善保存3d-21d內(nèi)紅細(xì)胞變形性指數(shù)，14d-21d內(nèi)的剛性指數(shù)；不同濃度的NO對紅細(xì)胞聚集性指數(shù)均沒有影響。高于相應(yīng)的最佳NO濃度范圍的時，紅細(xì)胞變形指數(shù)受損。 結(jié)論：懸浮紅細(xì)胞隨著保存時間的延長，紅細(xì)胞變形能力會逐漸降低。NO對庫血紅細(xì)胞變形功能具有調(diào)控作用，，通過補(bǔ)充最佳濃度的NO（如5μM）時，會使紅細(xì)胞變形性增加；儲存期間紅細(xì)胞的損傷與NO流失有關(guān)，適當(dāng)補(bǔ)充L-精氨酸（NO前體）可以恢復(fù)紅細(xì)胞的變形性。
[Abstract]:Nitric oxide (no), as a highly reactive free radical, has a wide range of physiological functions in vivo. No forms SNO-HbN by binding to the conserved Hb93Cys residue in erythrocyte hemoglobin (Hb). However, no in HB molecule can transfer from heme to thiol group, regulate the storage and release of no, and mediate vasodilation. With the prolongation of storage time, the red blood cells will lose a lot of no, and the physiological activities such as vasodilation, deformability and oxygen-carrying capacity of hemoglobin will decrease after being injected into the body. Therefore, it is of great theoretical significance and clinical value to understand the loss of no in the course of erythrocyte storage and to understand the mechanism of no in circulatory system in order to ensure the safety and effectiveness of erythrocyte transfusion. The aim of this study was to improve the morphology and function of suspended red blood cells by supplementing no in vitro, and to explore the mechanism of no on the morphology and storage damage of red blood cells, so as to provide a theoretical basis for the effective preservation of red blood cells. Objective: to add different concentrations of no reagent: L-Arginine / L arginine / no precursor to suspended red blood cells during preservation, and to analyze the changes of no content and erythrocyte deformability in different phases of suspension erythrocyte preservation. The aim of this study was to investigate the effect of no on the deformability of erythrocytes and evaluate the storage quality of erythrocytes. Methods: 10 cases of fresh suspended red blood cells were stored at 4 鈩

本文編號：1989212