脂多糖诱发的同基因妊娠BALB/c和NOD/SCID小鼠早产模型

目的：研究脂多糖（LPS）诱发同基因妊娠BALB/c小鼠和非肥胖性糖尿病/重度联合免疫缺陷（NOD/SCID）小鼠早产的机制。方法：在预先阻断或未阻断Toll样受体4（TLR4）的条件下采用LPS刺激,并比较各组BALB/c和NOD/SCID小鼠的早产率和胚胎死亡率。由于预实验显示预期的早产均发生于孕16d,因此,实验中在早产发生之前处死小鼠,收集每只孕鼠的胎盘。采用流式细胞术检测胎盘CD45^＋细胞表面TLR4、CD80和细胞内TNF-α的表达率。结果：采用LPS可诱发BALB/c小鼠早产,而NOD/SCID小鼠则对LPS的诱导有抵抗。经LPS刺激后,TLR4的表达在BALB/c和NOD/SCID小鼠均无显著改变,但是两组小鼠CD45^＋CD80^＋细胞的百分率均升高。相反,LPS刺激后仅BALB/c小鼠CD45＋TNF-α＋细胞的百分率升高,而NOD/SCID小鼠则否。通过预先阻断TLR4的表达可消除LPS对BALB/c小鼠CD80和TNF-α表达的影响,并显著降低LPS诱发的早产率。结论：虽然LPS未能改变TLR4的表达,但是二者相互作用,可激发CD45^＋CD80^＋细胞的动员,导致炎性细胞因子产生增多,并最终导致早产。BALB/c和NOD/SCID小鼠对LPS刺激的敏感性存在差异,提示NOD/SCID小鼠缺乏功能正常的T细胞和NK细胞,可能是这种小鼠对LPS诱发的早产有抵抗的原因之一。

Premature delivery induced by LPS in syngenetically impregnated BALB/c and NOD/SCID mice

AIM: To extend understanding of the mechanism of lipopolysaccharide (LPS)-induced preterm delivery in syngenetically impregnated BALB/c and NOD/SCID (nonobese diabetic/severe combined immunodeficiency) mice. METHODS: Strategies of LPS stimulation were pursued with or without previous Toll-like receptor 4 (TLR4) blocking. The incidence of LPS-induced preterm delivery and fetal death were compared between the BALB/c and NOD/SCID groups. Guided by the time when all expected preterm deliveries have occurred in the first experiment (i.e., day 16 of gestation), the LPS-stimulated mice, with or without previous TLR4 blocking, were killed at the beginning of preterm labor and pooled placentas were collected in each mouse in the second experiment. The expression of cell surface TLR4, CD80, and intracellular TNF-alpha in placenta CD45(+) cell population was determined by flow cytometry(FCM). RESULTS: It displayed that preterm delivery could be induced by LPS in BALB/c mice, while the NOD/SCID mice seemed to be resistant to LPS induction. Upon LPS stimulation, TLR4 expression was not changed either in BALB/c or in NOD/SCID mice, but the CD45(+) CD80(+) cell percentage was elevated in both groups. However, the CD45(+) TNF-alpha(+) cell percentage was increased merely in BALB/c mice after stimulation, while no such trend was observed in NOD/SCID mice. In BALB/c mice, the effect of LPS on CD80 and TNF-alpha expression could be abrogated by previous TLR4 blocking, which subsequently prevented LPS-induced preterm delivery. CONCLUSION: Although LPS do not alter TLR4 expression, it interacts with this receptor, triggers the mobilization of CD45(+) CD80(+) cells, results in elevated production of inflammatory cytokines, and finally results in preterm delivery. The diversity of sensitivity to LPS induction observed in BALB/c and NOD/SCID mice implies that the lack of functional T and NK cells in the NOD/SCID may be the reason why the NOD/SCID appeared to be resistant to LPS-induced premature labor.