血小板活化因子受体拮抗剂对幼年大鼠肠黏膜上皮细胞间紧密连接蛋白的影响  

Effect of platelet activating factor receptor antagonist on tight junction associated protein between epithelial cells of intestinal mucosa in young rats

作　　者：王丽杰[1] 刘春英 许玲芬[1] 高红[3] 姜卫国[4] 孙梅[1] 

机构地区：[1]中国医科大学附属第二临床学院儿科,辽宁省沈阳市110004 [2]解放军463医院儿科,辽宁省沈阳市110042 [3]中国医科大学附属第二临床学院中心实验室,辽宁省沈阳市110004 [4]中国医科大学附属第二临床学院病理科,辽宁省沈阳市110004

出　　处：《世界华人消化杂志》2006年第4期392-397,共6页World Chinese Journal of Digestology

摘　　要：目的:探讨血小板活化因子(platelet activating factor,PAF)受体拮抗剂对肠黏膜上皮细胞间紧密连接蛋白ZO-1的影响．方法:18日龄Wistar大鼠,随机分为对照组,内毒素组(LPS组)和PAF受体拮抗剂组(预防组和治疗组)．LPS组和对照组分别腹腔注射内毒素 (5 mg/kg)和生理盐水(1 mL/kg)．预防组和治疗组分别于每一时相点注射LPS前、后30 min 腹腔注射PAF受体拮抗剂BN52021(5 mg/kg)．按时间点分别处死动物,取回肠用于电镜观察,免疫组化及RT-PCR检测ZO-1．结果:电镜下对照组肠微绒毛及细胞间紧密连接未见异常．实验组上皮细胞连接增宽;微绒毛变细、稀疏,部分断裂、脱落;细胞器受损．拮抗剂组改变较实验组减轻．紧密连接蛋白ZO-1正常时均匀一致地分布于小肠上皮细胞连接处的尖端,呈蜂巢状,实验组ZO-1 分布不均,染色变淡．免疫组化平均光密度值及RT-PCR结果可见LPS组ZO-1表达明显低于对照组,6 h表达最低,光密度从对照组 0．224 7降至LPS组0．198 5,ZO-1 mRNA从1．18 降至0．16(P<0．01)．预防组及治疗组变化趋势同LPS组,6 h预防组及治疗组光密度分别为 0．199 2和0．203 8,ZO-1 mRNA分别为0．47和 0．53,与对照组相比均有显著差异(P<0．01)．各时相点ZO-1较LPS组高,预防组较治疗组 ZO-1略低,无统计学差异．结论:PAF可降低肠道紧密连接蛋白ZO-1,从而损害肠道的屏障功能,而PAF受体拮抗剂可减轻肠道屏障功能损伤的程度．AIM： To investigate the effect of platelet activating factor （PAF） receptor antagonist on the tight junction associated protein ZO-1 （zonula occludens-1） between epithelial cells of intestinal mucosa in young rats. METHODS： Eighteen day-old Wistar rats were randomly assigned into lipopolysaccharide （LPS） group, PAF receptor antagonist （prevention and treatment） group and control group. The rats in LPS and control group were intraperitoneally injected with LPS （5 mg/kg） and normal saline （1 mL/kg）, while the rats in prevention and treatment group were intraperitoneally injected with PAF receptor antagonist BN52021 （5 mg/kg） 30 min before and after LPS injection. Terminal ileum of each rat was collected for transmission electron microscopy. The expression of ZO-1 was determined by immunohistochemistry and reverse transcription polymerase chain reaction （RT-PCR） at both protein and mRNA level. RESULTS： Microvilli and tight junctions were intact in control group. Enlargement of tight junctions were observed in LPS group and microvilli were thin, rare or disrupt, and shed. The rough endoplasmic reticulum, mitochondria, and glycogen particles were damaged. The above changes were alleviated in PAF receptor antagonist group. The staining of ZO-1 in the control rats was similar to a honeycomb, which reflected a continuous and uniform distribution localized at the apical portion of cell-to-cell contact of the enterocytes. In LPS group, the signals of ZO-1 were disrupted and irregularly distributed at the outer enterocyte periphery. The content of ZO-1 was obviously lower in LPS group than that in the control group at 6 h （optical density： 0.198 5 ± 0.015 9 vs 0.224 7 ± 0.021 0, P 〈 0.01; mRNA： 0.16 ± 0.02 vs 1.18 ± 0.09, P 〈 0.01）. The content of ZO-1 was also significantly decreased in prevention and treatment group in comparison with that in control group at 6 h （optical density： 0.199 2 ± 0.008 7, 0.203 8 ± 0.006 7 vs 0.224 7 ± 0.021 0, P 〈 0.01; mRNA： 0.4

关 键 词：血小板活化因子 受体拮抗剂 屏障 紧密连接 ZO-1 

分 类 号：R725.7[医药卫生—儿科]