不同吸氧方式对新生鼠视网膜血管发育的影响

Effects of different oxygen inhalation modes on retinal vessels development in neonatal mice

目的吸氧是早产儿视网膜病的高危因素之一,目前吸入氧浓度、吸氧方式等与早产儿视网膜病的关系仍有争议。该实验通过研究不同吸氧方式对新生鼠视网膜血管发育的影响,为早产儿临床合理用氧提供实验依据。方法选择7日龄C57BL/6 J新生小鼠144只,随机分为6组,每组24只。实验组1:吸入氧浓度(F iO2)由30%逐渐升高至75%后突然中断吸氧;实验组2:F iO2由75%逐渐降低直至停氧;实验组3:吸75%高氧5天后骤然停氧;实验组4:吸75%高氧5天后逐渐降低F iO2直至停氧;实验组5:第1天吸75%高氧,第2天吸空气氧,交替进行,连续6 d;对照组:置于同实验条件下的空气中。实验组1,2,3,5和对照组分别于P12(生后第12天),P14,P17视网膜铺片(ADP酶染色法),实验组4于P14,P17,P22视网膜铺片,各组均在P17行视网膜切片(苏木素-伊红染色法)观察视网膜血管的增生和发育情况。结果①视网膜铺片显示:对照组:P12见少量无灌注区,周边血管结构清晰,P14无灌注区消失,血管形态基本正常,P17血管形态全部正常。实验组1,3,5:P12视网膜中央部有大片无灌注区,血管收缩、闭塞,P14新生血管开始形成,P17大量新生血管形成。实验组4:P14视网膜中央部大片无灌注区,有少量新生血管形成,P17无灌注区缩小,血管走行基本正常,深层血管有阻塞,P22血管发育基本正常。实验组2:铺片显示视网膜血管形态与对照组相似。②视网膜切片显示:实验组1,2,3,4,5小鼠突破视网膜内界膜的新生血管内皮细胞核的数目分别为49.50±1.36,5.17±0.67,47.68±4.70,5.74±2.37,29.15±2.48个,对照组为1.22±0.20个。实验组1,3,5与对照组相比差异均有显著性(P<0.05),实验组2,4与对照组相比差异无显著性(P>0.05)。结论不同吸氧方式对视网膜血管发育产生不同的影响,吸入氧浓度波动较大、吸入高浓度氧后突然停氧可严重影响新生鼠视网膜血管的发育,产生类似早产儿视网膜病的病变。因此临床上应尽早稳定早产儿的生命体征,减少氧疗,严密监测用氧情况,采取逐渐降低氧浓度的吸氧方式,并尽量避免氧浓度波动过大。

Objective This study was designed to investigate the effects of different oxygen inhalation modes on retinal vessels development in neonatal mice in order to provide experimental data for proper oxygen therapy for premature infants. Methods A total of 144 postnatal day （P） 7 C57BL/6J mice were randomly assigned into 6 groups according to different oxygen inhalation modes （n = 24）. Experimental group 1 was exposed to 30%, 40% , 50%, 60% and 75% oxygen in turn for one day respectively, followed by room air exposure for 5 days. Experimental group 2 was exposed to 75% , 60%, 50%, 40% and 30% oxygen in turn for one day respectively, followed by room air exposure for 5 days. Experimental group 3 was exposed to 75% oxygen for 5 days, followed by room air exposure for 5 days. Experimental group 4 was exposed to 75% oxygen for 5 days, 50% oxygen for 2 days and 30% oxygen for 2 days, then room air exposure for 6 days. The supplemental 75% oxygen and room air recovering was performed alternately for the mice in Experimental group 5 for 3 times and then room air exposure for 5 days. The Control group was exposed to room air for consecutive 10 days. The retinal vascular development and proliferation were evaluated by the retinal flat-mounts （ADPase stained retina） and cross-section. Results The peripheral vascular pattern was clear, and a few avascular areas were seen in the Control group at P12. At P14 the avascular area disappeared. At P17, the entire vascular pattern became completely normal. In the Experimental groups 1, 3 and 5, the central vessels became tortuous and constricted and the central avascular area increased at P12. At P14, neovascularization was seen, peaking at P17 in the Experimental groups 1, 3 and 5. In the Experimental group 4, the central avascular area increased and neovascularization was seen at P14, but the central avascular area was reduced mad abnormal neovascularization disappeared, with slight constriction of the deep vessels, at P17. Five days later the vascular pattern became almost normal in the Experimental group 4. The retinal vascular form of the Experimental group 2 was similar to that of the Control group. The average number of neovascular nuclei extending into the vitreous per cross-section in the Experimental groups 1,2, 3, 4, and 5 and the Control group was 49.50±1.36, 5.17±0.67, 47.68±4.70, 5.74±2.37, 29.15±2.48, and 1.22±0.20 respectively. There were significant differences between the Experimental groups 1, 3, 5 and the Control group （ P〈0.05 ）. Conclusions The effects of different oxygen inhalation modes on the retinal vessels development in neonatal mice were different. The obvious fluctuation of inhaled oxygen concentration and abrupt stop of supplemental oxygen after high levels of supplemental oxygen may severely affect the development of retina vascular, leading to the pathologic changes similar to retinopathy of prematurity.