束缚应激大鼠血清淋巴细胞转化抑制因子的研究

A STUDY ON SERUM SUPPRESSIVE FACTOR(S) ON LYMPHOCYTE PROLIFERATION IN RATS UNDER RESTRAINT STRESS

为研究应激对淋巴细胞转化的影响,将 SD 大鼠四肢束缚于支架上,仰卧位,室温(20℃)下维持20h,对照组留置原饲养笼中,不予惊动。然后在乙醚轻麻下穿刺心脏取血,肝素化后密度梯度离心分离淋巴细胞,或待凝后分离血清。结果表明,应激大鼠外周血淋巴细胞对刀豆素(Con A)诱导的转化反应明显下降(p<0.01,n=8,ANOVA),而且应激大鼠血清可明显抑制正常小鼠淋巴细胞转化,这提示应激大鼠血清中可能存在某种具有抑制淋巴细胞转化的活性物质。进一步的分析实验表明,这种血清经加热56℃(30min),30%甲醇或透析(透析袋孔径阻滞分子量为6000)处理,抑制活性均不受影响;但经加热100℃(3min),80%甲醇或胰蛋白酶(64/μg/ml)处理,抑制活性丧失。提示这种抑制活性物质很可能是一类蛋白质。

In order to study the effect of stress on lymphocyte proliferation,SD rats wererestrained with four limbs tied on a frame in supine position at room temperature(20℃)for 20h,and control animals were not disturbed in home cage.The bloodwas then collected from the heart under light ether anesthesia.The peripheralblood lymphocytes were separated from heparinized whole blood by density gra-dient(d 1.077)centrifugation,or the serum was obtained after the blood coagulatedat 4℃ for about 6h.It was found that the blood Iymphocyte proliferation indu-ced by Con A was significantly inhibited in the stressed group as compared withthe control(P<0.01,n=8,ANOVA).The result was in accordance with ourearlier study in which the animals were stressed with electric shock.In the pre-sent study,it was also found that the serum of the stressed animals was capable ofsuppressing Con A-induced lymphocyte proIiferation of normal mice(P<0.01,n=8,ANOVA)to a significant extent.Thus the present experiment suggests thatthere is some substance with suppressive activity on lymphocyte proliferation in theserum of the stressed rats.The serum lost its suppressive activity when it washeated to 100℃(3 min),treated with 60% methanol or incubated with trypsin(64 μg/ml),thus suggesting that the suppressive factor(s)most likely is a kind ofprotein.