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目的探讨胎膜早破(PROM)相关性胎儿感染胎肺组织中人正五聚蛋白3(PTX3)mRNA的表达及临床意义。方法收集2010年10月-2011年10月在首都医科大学附属北京妇产医院因死胎、死产、引产或流产的胎肺组织标本12份。将PROM并肺部感染的胎儿肺组织6份为观察组,无PROM、无肺部感染的胎儿肺组织6份为对照组。对留取肺组织行40 g.L-1多聚甲醛固定,2h内石蜡包埋,4μm连续切片。行HE染色,以普通光学显微镜观察其肺组织的病理学变化。提取储存于液氮中新鲜肺组织的总RNA,反转录为cDNA,琼脂糖电泳明确扩增的DNA与目标DNA分子大小是否相同,提取成功后,采用SYBR GreenⅠ实时荧光定量(qRT)-PCR和相对定量分析方法,以GAPDH基因为内参基因,检测2组胎儿肺组织中PTX3 mRNA的表达,采用相对定量△Ct法计算mRNA相对表达量。结果 1.肉眼所见观察组胎儿肺体积较小、颜色暗红,而对照组胎儿肺组织颜色粉红,体积饱满,肺组织切面出血量较少。2.光学显微镜下,观察组胎肺组织内可见大量中性粒细胞浸润、黏性液体和炎性细胞渗出,大量红细胞漏入;对照组上述病理改变则较轻。3.qRT-PCR检测结果显示,PTX3 mRNA在2组胎儿肺组织中均有表达,观察组△Ct值为5.81±0.63,对照组△Ct值为4.20±0.70,2组比较差异有统计学意义(t=4.17,P=0.002)。结论 PTX3 mRNA在PROM相关性胎儿感染的胎肺组织中的表达明显增加,可作为判断胎儿肺感染的早期敏感指标之一。
Objective To investigate the expression of PTX3 mRNA in fetal lung tissue associated with premature rupture of membranes (PROM) and its clinical significance. Methods Twelve fetal lung tissue samples were collected from stillbirth, stillbirth, induced abortion or abortion at Beijing Maternity Hospital affiliated to Capital Medical University from October 2010 to October 2011. 6 cases of PROM and lung infection of fetal lung tissue as the observation group, no PROM, no pulmonary infection in 6 cases of fetal lung tissue as control group. The lung tissue was collected fixed 40 g.L-1 paraformaldehyde, paraffin embedded within 2h, 4μm serial sections. HE staining, the optical microscope to observe the pathological changes of lung tissue. Total RNA was extracted from fresh lungs in liquid nitrogen and reverse transcribed into cDNA. Whether the DNA amplified by agarose gel electrophoresis was the same as the size of target DNA was determined by SYBR Green Ⅰ real-time fluorescence quantitative PCR (qRT) -PCR And relative quantitative analysis method, GAPDH gene was used as an internal reference gene to detect the expression of PTX3 mRNA in two groups of fetal lung tissues, and the relative quantitative △ Ct method was used to calculate the mRNA relative expression. The results of 1 eye observation group fetal lung volume smaller, dark color, while the control group of fetal lungs pink color, full size, less blood loss in lung tissue section. Under optical microscope, a large number of neutrophil infiltration, viscous fluid and inflammatory cells exudation were observed in the fetal lung tissue in the observation group, and a large amount of erythrocyte leakage was found. In the control group, the pathological changes were lighter. The results of qRT-PCR showed that PTX3 mRNA was expressed in both groups of fetal lung tissue, the △ Ct value of the observation group was 5.81 ± 0.63, the △ Ct value of the control group was 4.20 ± 0.70, the difference was statistically significant (t = 4.17, P = 0.002). Conclusions The expression of PTX3 mRNA in fetal lung tissues infected with PROM-related fetuses is significantly increased, which may be used as one of the early sensitive indicators for determining fetal lung infection.