束缚应激对Pg-LPS诱导的大鼠腹腔巨噬细胞释放细胞因子的影响

Effects of restain stress and Pg-LPS on the change of cytokines secretion from peritoneal macrophages in rats

目的:检测束缚应激对牙龈卟啉菌内毒素(Pg-LPS)诱导的大鼠腹腔巨噬细胞释放IL-Iβ、IL-2、IL-6、IL-10等细胞因子的影响,初步探讨应激影响牙周疾病的机制。方法:采用束缚应激方式,将大鼠随机分为正常对照组、应激组,于应激结束时处死,常规收集腹腔液。巨噬细胞于贴壁纯化后用RPMI-1640培养液,RPMI-1640培养液+1μg/mL Pg-LPS继续培养。于24 h后分别收集上清,用Luminex100检测仪测定上清液中IL-Iβ、IL-2、IL-6、IL-10等的水平。结果:①12 d应激组大鼠腹腔巨噬细胞数量较无应激组及1 d应激组显著下降。②1天应激组各细胞因子水平与无应激组比较均无显著差异。12 d应激组的IL-1β水平比无应激组显著增加(P<0.05)、IL-6水平比无应激组显著降低(P<0.05);IL-2水平比1天应激组显著降低(P<0.05)。IL-10在各组间差异无统计学意义(P>0.05)。结论:慢性束缚应激可引起大鼠腹腔巨噬细胞总量减少;慢性束缚应激可改变应激宿主对牙龈卟啉单胞菌感染的反应,这可能是慢性应激引起牙周疾病的机制之一。

Objective： To explore the effects of restrain Pg-LPS and stress on the change of cytokines secretion from peritoneal macrophages in rats. Method： Rats （SD, male） were tested by using restrain stress. Rats were divided into 3 ex- perimental groups randomly （no stress, one day stress, 12 days stress）.After stress conditions,the rats were sacrificed, and their peritoneal macrophages were isolated, cultured and purify. All the cells were then cultured with RPMI-1640＋1 Ixg/ mL Pg-LPS. After 24 hours, cell culture supernatants were collected and assayed for IL-I[~,IL-2,IL-6,IL-10. Result： The number of cells in the peritoneal cavity of 12 days stressed rats was significantly reduced in comparison to those from non- stressed animals （P 〈0.05）. The level of cytokines was no significant difference between the one day stress and no stress group （P 〉0.05）. The level of IL-113,IL-6 is suggested significant difference between the 12 days stress and no stress group （19 〈0.05）. The level of IL-2 is suggested significant difference between the 12 days stress and 1 day stress group （P 〈 0.05）. The level of IL-10 is suggested no significant difference within groups （P 〉0.05）. Conclusion： The results suggest chronic restrain stress can reduce the number of macrophages in the peritoneal cavity in rats. Chronic restrain stress modu- lates cytokine secretion from Pg- LPS stimulated macrophages.