大鼠中耳上皮细胞形态学变化与杀菌-通透性增强蛋白的关系(英文)

Relation between the morphologic change of rat middle ear epithelium and bactericidal/permeability increasing protein

背景:中耳形态学的恢复是慢性分泌性中耳炎好转的重要指标。寻找有效的中和脂多糖,减轻脂多糖对中耳黏膜的毒副作用的药物已被引起关注。目的:研究杀菌-通透性增强蛋白(bactericidal/permeability-increasingpro-tein,BPI)中和脂多糖从而抑制脂多糖导致的中耳上皮细胞形态的改变。设计:非随机、对照的实验研究。地点和对象:实验地点为吉林大学。对象:清洁级纯种Wistar大鼠,雌雄不限,体质量200～250g,购自吉林大学实验动物中心,饲养环境温度20～25℃,湿度60%～70%。干预:将10只正常Wistar大鼠的中耳上皮细胞放置于24孔多聚赖氨酸培养板中培养7d后,分为3组:未治疗组:在没有脂多糖和BPI的培养板中继续培养;脂多糖组:将1μg/L脂多糖加入培养板中培养;BPI治疗组:在1μg/L脂多糖加入培养板培养7d后再加入1mg/LBPI继续培养。主要观察指标:生长4周后,光镜下观察细胞形态学变化。结果:与未治疗组比较,脂多糖组在1周后表现为细胞肿胀、变大,同时数量增多,极少数区域出现细胞破裂现象;2周时细胞明显增大伴有细胞破裂现象,部分细胞死亡,伴有纤毛细胞减少,分泌细胞增多。4周时细胞破裂增多,细胞死亡加剧。而BPI治疗组2周时肿胀的细胞数目及细胞破裂现象远不及脂多糖组2周时严重,4周后表现为细胞形态正常。

BACKGROUND:The recovery of middle ear morphology is an important index of improvement of chronic secretory otitis media.Looking for effective medicine to neutralize lipopolysaccharide and relieving the virulence of lipopolysaccharide to middle ear mucosa have drawn more attention. OBJECTIVE:To study the effect that bactericidal/permeability increasing protein(BPI) neutralizing lipopolysaccharide to inhibit morphologic changes of the middle ear epithelium due to lipopolysaccharide. DESIGN:A nonrandomized controlled experimental study. SETTING and PARTICIPANTS:The setting is in Jilin University.Participants were 10 purebred Wistar rats in clean grade,weighing 200-250 g,purchased from Center for Experimental Animals,Jilin University,and feeding condition was at 20-25 ℃,humidity of 60%-70%. INTERVENTIONS:The middle ear epithelial cells were obtained from 10 Wistar rats and distributed into 24 well tissue culture plate coated with poly lysine.These epithelial cells were cultured for 7 days,and then divided into 3 groups:Untreated group was cultured without lipopolysaccharide and BPI in the culture medium;Lipopolysaccharide group was cultured with 1 μg/L lipopolysaccharide in the culture medium;BPI group was cultured with 1 μg/L lipopolysaccharide for 7 days,and then with added 1 mg/L BPI in the culture medium. MAIN OUTCOME MEASURES:The morphologic changes of the middle ear epithelial cells were observed under light microscope after 4 weeks. RESULTS:Compared to untreated group,lipopolysaccharide group showed that many cells became greater and swollen,and the number of them was increasing,the phenomena of cell breakage were observed in few areas 1 week later.At 2 weeks,cells became much greater and swollen,and the number of broken and dead cells was obviously increased.Moreover,an increase in secretory cells and a decrease in ciliated cells were observed.After 4 weeks,the broken and dead cells distinctly were increased.However,at 2 weeks of BPI group,the increasing number of swollen and broken cells was much less significant than that of lipopolysaccharide group at 2 weeks.After 4 weeks in culture medium with BPI,cells maintained relatively normal gross morphology,with few swollen cells and broken cells,and ciliated cells increased,secretory cells decreased. CONCLUSION:BPI can bind effectively to lipopolysaccharide,inhibit dysfunction caused by lipopolysaccharide,and recover normal morphology of the middle ear epithelium.It also can reduce the occurrence of complications and sequelae of secretory otitis media.