本文選題：反應性氮代謝物 + 反應性氧代謝物　； 參考：《福建醫(yī)科大學》2008年碩士論文

【摘要】： [目的]:探討反應性氮代謝物(RNM)對NK細胞抗K562細胞活性的影響及硫普羅寧和還原型谷胱苷肽逆轉(zhuǎn)ROM、RNM抑制NK細胞抗K562細胞活性的免疫佐劑作用。 [方法]:1.在NK細胞、K562細胞培養(yǎng)體系中,加入外源性ONOO-,觀察RNM和ROM產(chǎn)量、NK細胞活性變化;2.采用IL-2及PHA為激活劑,使RNM、ROM產(chǎn)量增多,觀察KIR和NK細胞活性的相應變化,然后在MO+NK+K562(E/T=10/1,E/MO=10/2)細胞培養(yǎng)體系中加入不同濃度的硫普羅寧、還原型谷胱苷肽和二氫氯組胺,定期檢測RNM、ROM產(chǎn)量、KIR及IL-6、TNF-β、IFN-γ的產(chǎn)量。 [結(jié)果]:1.在NK細胞、K562細胞培養(yǎng)體系中,加入外源性ONOO-后,K562細胞數(shù)量和NK細胞活性顯著降低,而RNM、ROM產(chǎn)量增加。2.①在MO細胞培養(yǎng)體系中,隨著MO細胞數(shù)量的增加,RNM、ROM產(chǎn)量有所上升;而經(jīng)IL-2/PHA激活后,在MO=10組, RNM從81.8898±0.7874umom/l升至91.0308±0.8956umom/l , ROM從193.1685±9.6505U/ml升至356.4275±5.9642U/ml (P0.05)。②在NK+K562混合培養(yǎng)體系中,加入IL-2/PHA后,當E/T =10/1時,RNM略有增加,而ROM從58.6326±2.5141U/ml增加到141.915±8.2593U/ml(P0.05),KIR從79.98%升至93.29%(P0.05);當按E/MO=10/2、10/5、10/10三種比例加入MO細胞后,RNM、ROM產(chǎn)量隨著MO細胞數(shù)量的增加而增加,而IL-6、TNF-β、IFN-γ的產(chǎn)量和KIR則相反。對K562細胞數(shù)量作偏相關(guān)分析,RNM與KIR的負相關(guān)系數(shù)大于ROM(分別為r= -0.8511,-0.7141)。③在IL-2/PHA+NK+MO+K562混合培養(yǎng)體系中,加入TIP、GSH、DHT后,RNM產(chǎn)量從73.7390±3.7908 umom/l分別降至64.5516±2.1570umom/l、63.0222±2.3090 umom/l、73.1052±1.4528 umom/l,ROM產(chǎn)量從321.5488±10.5030 U/ml分別降至55.1665±7.0950 U/ml、54.6136±5.9515 U/ml、108.6608±6.2110U/ml,而KIR則從65.78%分別上升至84.47%、84.77%、79.56%。隨著各藥物濃度的增加,RNM、ROM產(chǎn)量逐漸減少,而IL-6、TNF-β、IFN-γ產(chǎn)量和KIR則逐漸升高,KIR與RNM、ROM產(chǎn)量呈明顯負相關(guān)(P0.01)。在清除RNM、ROM,提高NK細胞抗K562細胞活性方面,硫普羅寧與還原型谷胱苷肽作用相似(P0.05),但均優(yōu)于二氫氯組胺(P0.05)。 [結(jié)論]:1.外源性ONOO-對NK、K562細胞有毒性,并可轉(zhuǎn)化及誘導細胞產(chǎn)生ROM。2.MO細胞是ROM、RNM的最主要來源,而K562細胞、NK細胞也產(chǎn)生少量的ROM、RNM.。3.RNM、ROM均可使NK細胞的抗瘤(抗K562)活性下降。4.硫普羅寧、還原型谷胱苷肽均可通過清除ROM、RNM保護NK細胞,提高NK細胞抗瘤活性,且對ROM、RNM的清除表現(xiàn)出一定的量效關(guān)系,但二氫氯組胺不能清除RNM。5.在清除ROM、RNM及提高NK細胞對K562細胞的抑制率方面,硫普羅寧、還原型谷胱苷肽效應相當,但均優(yōu)于二氫氯組胺,且毒副作用較低,有望成為新的免疫佐劑用于白血病的過繼性免疫治療中。
[Abstract]:Objective: to investigate the effects of reactive nitrogen metabolites (RNM) on the anti-K562 cell activity of NK cells and the effect of tipronine and reduced glutathione on reversing the inhibitory effect of RNM on NK cell anti-K562 cell activity. [method]: 1. In K562 cell culture system, exogenous ONOO was added to observe the change of NK cell activity in RNM and ROM production. Using IL-2 and PHA as activators, the production of RNMN ROM was increased, the corresponding changes of KIR and NK cell activity were observed, and then different concentrations of tiopronin, reduced glutathione and dihydrochlorohistamine were added to MO NK K562 cell culture system. The yield of RNMN ROM and the yield of IL 6 TNF- 尾 and IFN- 緯 were detected regularly. [result]: 1. In K562 cell culture system, the number of K562 cells and the activity of NK cells decreased significantly after the addition of exogenous ONOO-, while RNMN ROM production increased .2.1 in MO cell culture system, and the ROM production increased with the increase of MO cell number. After activation of IL-2 / PHA, in MO10 group, RNM increased from 81.8898 鹵0.7874umom/l to 91.0308 鹵0.8956umom/l, ROM from 193.1685 鹵9.6505U / ml to 356.4275 鹵5.9642U / ml (P0.05). In NK K562 mixed culture system, after adding IL-2 / PHA, RNM increased slightly when E / T was 10 / 1. However, the ROM increased from 58.6326 鹵2.5141U / ml to 141.915 鹵8.2593U / ml (P0.05) from 79.98% to 93.29% (P0.05). When added to MO cells according to the ratio of E / MO10 / 10 / 210 / 5 / 10 / 10, the ROM output increased with the increase of MO cell number, whereas the output of IL-6TNF- 尾 -IFN- 緯 increased with the increase of MO cell number, whereas the yield of IL-6TNF- 尾 -IFN- 緯 increased with the increase of MO cell number. The negative correlation coefficient between RNM and KIR in K562 cells was higher than that in ROM (r = -0.8511-0.7141) in IL-2 / PHA NK MO K562 co-culture system. 鍔犲叆TIP,GSH,DHT鍚，

本文編號：2081489