卵泡抑素样蛋白1对先天性膈疝大鼠模型中肺泡上皮细胞分化的影响  

Effects and significance of follistatin-like 1 on alveolar epithelial cell differentiation in a congenital diaphragmatic hernia rat model

作　　者：张杰 黄璜[1] 覃智杰 王培然 周洲[1] 范霞[1] 罗淼[1] 唐应明[1] 何国庆[1] 李飞[1] Zhang Jie;Huang Huang;Qin Zhijie;Wang Peiran;Zhou Zhou;Fan Xia;Luo Miao;Tang Yingming;He Guoqing;Li Fei(Department of Pediatric Surgery,Guizhou Provincial People's Hospital,Guiyang 550002,China)

机构地区：[1]贵州省人民医院小儿外科,贵阳550002

出　　处：《中华小儿外科杂志》2023年第7期641-647,共7页Chinese Journal of Pediatric Surgery

基　　金：贵州省卫生健康委科学技术基金(gzwjkj2019-1-001)。

摘　　要：目的探讨卵泡抑素样蛋白1(follistatin-like 1,FSTL1)对先天性膈疝(congenital diaphragmatic hernia,CDH)大鼠模型中肺泡上皮细胞分化的影响及意义。方法选取成年SPF级SD大鼠15只,10只受孕后按随机数字表法分为模型组(6只)及空白组(4只)。应用除草醚建立CDH大鼠模型,模型组中解剖发现膈疝的胎鼠定为CDH组,空白组中解剖未发现膈疝的胎鼠定为对照组。取CDH组及对照组左肺进行研究。通过HE染色比较两组肺组织形态学差异;应用免疫组织化学、蛋白质印迹法及qRT-PCR检测FSTL1、甲状腺转录因子1(thyroid transcription factor 1,TTF1)及水通道蛋白5(aquaporin 5,AQP5)在两组肺组织中的表达变化;应用蛋白质印迹法检测两组肺组织中表面活性蛋白B(surfactant protein B,SP-B)和表面活性蛋白C(surfactant protein C,SP-C)及其前体蛋白的表达水平。两组均数的比较采用独立样本t检验。结果成功建立CDH大鼠模型,获得CDH胎鼠21只。CDH组辐射状肺泡计数低于对照组[(2.88±0.19)比(5.68±0.24),P<0.001];免疫组织化学检测显示FSTL1、TTF1、AQP5在两组肺组织中均有表达,且在不同组别肺组织中表达强度不同。CDH组中FSTL1的mRNA及蛋白表达[(0.5550±0.0462)和(0.6949±0.0250)]低于对照组[(1.0092±0.1636)和(1.0062±0.1067)],差异有统计学意义(P<0.05)。CDH组中AQP5的mRNA及蛋白表达[(0.5524±0.1474)和(0.4892±0.1407)]低于对照组[(1.0103±0.1822)和(0.8352±0.1149)],差异有统计学意义(P<0.05)。相反,CDH组中TTF1的mRNA及蛋白表达[(2.8073±0.5710)和(0.7389±0.0660)]高于对照组[(1.0172±0.2225)和(0.3278±0.0353)],差异有统计学意义(P<0.05)。此外,CDH组中表面活性物质前体蛋白Pro-SP-B及Pro-SP-C的表达[(1.1570±0.1354)和(0.9588±0.0249)]较对照组[(0.9113±0.0349)和(0.4999±0.0672)]增高,而SP-B及SP-C的表达[(0.4587±0.0264)和(0.1344±0.0658)]较对照组[(0.7112±0.0668)和(0.3296±0.0296)]降低,差异均有统计学意义(P<0.05)。结论Objective To explore the effects and significance of follistatin-like 1(FSTL1)on alveolar epithelial cell differentiation in a congenital diaphragmatic hernia(CDH)rat model.Methods Adult specific pathogen-free(SPF)grade Sprague-Dawley(SD)postnatal rats were selected and randomized into two groups of model and blank.Nitrofen was employed for establishing a CDH rat model.Fetal rats were dissected.CDH group was defined as fetal rats with CDH in model group and control group normal fetal rats in blank group.Left lung of fetal rats was harvested for further researches.Lung morphological difference between two groups were compared after hematoxylin-eosin(HE)stain.The expression levels of FSTL1,thyroid transcription factor 1(TTF1)and aquaporin 5(AQP5)were examined by immunohistochemistry,Western blot and qRT-PCR.Western blot was utilized for detecting the expression levels of surfactant protein B,surfactant protein C and their protein precursors.All comparisons were conducted with independent t test.Results CDH rat model was successfully established(n=21).Radial alveolar count of fetal rat lungs in CDH group were lower than that in control group(2.88±0.19 vs 5.68±0.24)(P<0.001).Immunohistochemistry indicated that FSTL1,TTF1 and AQP5 were expressed in lung tissue and their expression intensities differed between two groups.The mRNA and protein expression of FSTL1 in CDH group(0.5550±0.0462 vs 0.6949±0.0250)were less than those of control group(1.0092±0.1636 vs 1.0062±0.1067)and the differences were statistically significant(P<0.05).The mRNA and protein expression of AQP5 in CDH group(0.5524±0.1474 vs 0.4892±0.1407)was lower than those of control group(1.0103±0.1822 vs 0.8352±0.1149)and there were statistically significant inter-group differences(P<0.05).Whereas,the mRNA and protein expression of TTF1 in CDH group(2.8073±0.5710 vs 0.7389±0.0660)was higher than that of control group(1.0172±0.2225 vs 0.3278±0.0353).These differences were statistically significant.Furthermore,the protein expressions of protein

关 键 词：疝 横膈 先天性 卵泡抑素样蛋白1 肺发育不良 肺泡上皮细胞 

分 类 号：R726.5[医药卫生—儿科] R-332[医药卫生—临床医学]