摘要
目的研究核黄素光化学方法对淋巴细胞的增殖活性及细胞因子分泌活性的抑制程度,观察该方法对淋巴细胞增殖周期的影响。方法实验分为实验组:随机选择捐献的全血将从中收集的淋巴细胞悬浮于核黄素终浓度为50μmol/L的1640缓冲溶液中,注入PVC透光转移袋内(440nm处透光率为80%),用400—500nm的可见光照射,照射量为8.8J/ml;对照组:为相同来源、悬浮于不含核黄素的1640缓冲溶液中、未接受光照处理的淋巴细胞。以植物血凝素同步刺激2个组的淋巴细胞,用MTS非放射性细胞增殖试验检测淋巴细胞的增殖活性,计算核黄素光化学处理对淋巴细胞的增殖抑制率;用酶标方法检测细胞因子;用流式细胞术观察经植物血凝素(PHA)刺激后的淋巴细胞周期变化(RPT-lymphocytes)。结果与对照组比较,实验组经过核黄素光化学法处理后的淋巴细胞(RPT-lymphocytes)对PHA刺激的增殖抑制率为(99.21±1.06)%,淋巴细胞IL-1β、-2、-6、-8及TNF-α、IFN-γ细胞因子的分泌量比对照组细胞分别降低了(95.09±2.60)%、(98.20±1.64)%、(98.77±0.97)%、(92.30±11.04)%及(98.82±1.42)%、100%;实验组S期细胞占0.73%,对照组S期细胞则占32.5%。结论可见光激发的核黄素光化学方法可有效抑制淋巴细胞的增殖活性和细胞因子分泌活性,阻止淋巴细胞进入细胞增殖周期,提示该方法可能是预防TA-GVHD的一种有效并且可行的方法。
Objective To study the effects of riboflavin photochemical treatment （RPT） on the inhibition of lymphocyte proliferation and cytokine secretion, and also the effects on the cell cycles. Methods Lymphocytes collected from random blood donors and riboflavin with the final concentration of 50μM were injected into transparent PVC bags and then exposed to visible light ranging from 400nm to 500nm at a final dose of 8.8J/ml. The control lymphocytes didn＇ t receive the riboflavinphotochemical treatment. Both lymphocytes aforementioned were tested for the proliferative ability, the level of several cytokines and the change of cell cycle after the stimulation of phytohemagglutinin （ PHA ）. Results The proliferation inhibition rate of riboflavin photochemically treated lymphocytes （ RPT-lymphocytes ） after the stimulation was （99.21 ± 1.06）%, and the production of IL-1β, IL-2, IL-6, IL-8, TNF-α and IFN-γ was decreased by （95.09±2.60）%, （98.20±1.64）%, （98.77±0.97）%, （92.30±11.04）%, （98.82±1.42）% and 100%, respectively, compared to the controls. After 3-day PHA stimulation, 32.5% of the control lymphocytes were in S-phase, while 0.73% of the RPT-lymphocytes were in S-phase. Conclusions Riboflavin photochemical treatment with visible light could inhibit the lymphocyte proliferation, cytokine production, and also prevent the lymphocyte enter into cell proliferation cycle, suggesting that this method would be a promising method to prevent TA-GVHD.
出处
《中国输血杂志》
CAS
CSCD
2008年第2期100-103,共4页
Chinese Journal of Blood Transfusion
基金
上海市科学技术委自然基金(编号:06ZR141491)
上海市医学领先专业重点学科建设项目资助课题(编号:105Ⅲ003)
