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目的观察不同信号转导调控剂对严重烫伤小鼠脾脏T淋巴细胞分泌白细胞介素(IL)2、IL-10功能的影响,并探讨其机制。方法分离正常小鼠及高压热蒸气烫伤小鼠(烧伤面积18％TBSA,Ⅲ度)伤后12、96 h的脾脏T淋巴细胞。分为正常对照组(不加T淋巴细胞活化相关信号转导分子调控剂)、调控剂组[正常细胞分别加入以下调控剂:膜蛋白激酶C(PKC)抑制剂H-7(50μmol／L,1 ml)、PKC激活剂佛波酯(TPA,30μmol／L,1 ml)、非受体型蛋白酪氨酸激酶(PTK)抑制剂Herbimycin(10μmol／L,1 ml)、丝裂原活化蛋白激酶激酶(MAPKK)抑制剂PD098059(25μg／ml,1 ml)和Ca2+导入剂A23187(100 nmol/L,1 ml)]、烫伤组(不加调控剂)、烫伤+调控剂组(加入上述调控剂)。检测各组细胞IL-2、IL-10的分泌水平。结果伤后各组细胞中IL-2、IL-10含量均低于正常对照组(P<0．05或0．01);烫伤12 h+H-7组、烫伤96 h+H-7组分别低于烫伤12 h组和烫伤96 h组(P<0．01)。TPA能使IL-2、IL-10分泌明显升高,但对IL-2升高效能更显著。加入PD098059的各组IL-2、IL-10值均低于正常对照组(P<0．05或0．01)。Herbimycin能显著降低烫伤12 h+Herbi- mycin组及烫伤96 h+Herbimycin组IL-2的分泌,对IL-10亦有明显抑制。烫伤12 h+A21387组中IL-2、IL-10含量为(2417±39)、(2793±25)pg／ml,烫伤96 h+A21387组为(921±50)、(2633±35) pg／ml均明显高于烫伤12 h组[(1542±40)、(2390±15)pg／ml]和烫伤96 h组[(328±19)、(1618±21)pg／ml(P<0．05或0．01)]。结论PKC、Ca2+、MAPKK和PTK在严重烫伤小鼠脾脏T淋巴细胞分泌IL-2、IL-10的功能紊乱中起重要作用,其中PKC和PTK主要作用于IL-2的分泌而MAPKK影响IL-10的分泌。TPA和A21387均能显著提高烫伤后IL-2的分泌,并明显纠正伤后IL-2／IL-10分泌比值的失调。
Objective To observe the effects of different signal transduction regulators on the secretion of interleukin (IL) 2 and IL-10 by splenic T lymphocytes in severely scalded mice and to explore its mechanism. Methods Splenic T lymphocytes from normal mice and scalded mice (burn area 18% TBSA, grade Ⅲ) 12 and 96 hours after injury were isolated. Divided into normal control group (without T-lymphocyte activation related signal transduction molecular regulators), the regulator group [normal cells were added the following modulators: membrane protein kinase C (PKC) inhibitor H-7 (50μmol / L, 1 ml), PKC activator phorbol ester (TPA, 30 μmol / L, 1 ml), non-receptor tyrosine kinase (PTK) inhibitor Herbimycin (10 μmol / L, (25 nmol / L, 1 ml) and Ca2 + (100 nmol / L, 1 ml), scald (no regulators) and scald + regulators Agent). The secretion of IL-2 and IL-10 in each group was detected. Results The levels of IL-2 and IL-10 in the injured cells were lower than those in the normal control group (P <0.05 or 0.01), scalded for 12 h + H-7 and scalded for 96 h + H-7 Respectively, lower than scalded 12 h group and scalded 96 h group (P <0.01). TPA can make IL-2, IL-10 secretion was significantly increased, but more pronounced on the IL-2 increased. The levels of IL-2 and IL-10 in PD098059 group were lower than those in normal control group (P <0.05 or 0.01). Herbimycin can significantly reduce IL-2 secretion at 12 h + Herbi-mycin group and 96 h + Herbimycin group, and also inhibit IL-10 significantly. The contents of IL-2 and IL-10 in the burn group at 12 h and A21387 were (2417 ± 39) and (2793 ± 25) pg / ml, / Ml were significantly higher than those in scald at 12 h [(1542 ± 40), (2390 ± 15) pg / ml] and scald at 96 h (328 ± 19) and (1618 ± 21) pg / .05 or 0.01)]. Conclusions PKC, Ca2 +, MAPKK and PTK play an important role in the dysfunction of IL-2 and IL-10 secreted by splenic T lymphocytes in severely scalded mice. PKC and PTK mainly play roles in the secretion of IL-2 and MAPKK affects IL- 10 secretion. Both TPA and A21387 can significantly increase the secretion of IL-2 after scald and significantly correct the imbalance of IL-2 / IL-10 secretion ratio after injury.