血IL-6、8及MMP-9表达水平在宫内感染所致新生鼠肺损伤中的意义

发布时间：2018-04-01 22:10

本文选题：宫内感染　切入点：脂多糖　出处：《青岛大学》2017年硕士论文

【摘要】：目的:采用腹腔注射脂多糖(lipopolysaccharide,LPS)的方法构建稳定的宫内感染的孕鼠模型,通过肺组织病理切片了解宫内感染对肺部组织造成的损伤程度,监测新生鼠血中IL-6、IL-8、MMP9的浓度,探究前述炎症因子对预测肺部疾病的价值。方法:宫内感染组:孕18天的孕鼠18只,分别于宫腔内注射合适浓度的LPS(0.6、0.7、0.8、0.9、1.0mg/kg)。对照组:孕18天的孕鼠3只,于宫腔内注射和感染组LPS等量的灭菌生理盐水。查看自然分娩的新生大鼠存活数量,每只均擦干称重,再与分娩母鼠共同饲养,记录每天不同时刻新生大鼠的呼吸频率、动度以及有无发绀等异常情况;首先收集自然分娩后胎盘(自然分娩下收集胎盘容易失败,可选择择期剖宫产收集新鲜胎盘);按照1、3、7日龄分别随机选取两组中自然分娩所得的每窝新生大鼠中的8只新生大鼠,每组每个时间点共48份肺组织标本。将随机选取的新生大鼠断头取血(心脏取血或眼球内眦取血),留取血标本用于ELISA实验;处死后迅速取出肺组织,将所取的胎盘与肺均进行HE染色制成切片备用。比较实验组与对照组中随着药物剂量递减孕鼠死亡率、流产率以及新生大鼠死亡率的变化,比较不同LPS组新生鼠胎盘及肺病理评分、新生鼠体重/肺重、ELISA法监测胎鼠血中的IL-6、IL-8、MMP-9表达差异有无统计学意义。结果:(1)注射LPS浓度大于0.7mg/kg时,各组胎盘病理与对照组相比较差异有统计学意义(P0.05),胎盘病理可见存在炎症细胞开始浸润绒毛膜羊膜,且注射孕鼠LPS浓度越高,炎症细胞愈多,浸润组织部位越多。(2)通过注射不同浓度LPS造成肺部炎症浸润的程度也不相同,不同组炎症评分与对照组比较差异均有统计学意义(P0.05),提示宫内感染可造成肺部炎症,当注射LPS浓度越高,肺部组织炎症细胞浸润越多,肺部组织破坏越多,新生大鼠肺部损伤越重,并且这种损伤会持续至生后7天左右。(3)注射不同浓度LPS的新生大鼠每天呼吸情况及生长指标相比较,实验组与对照组新生大鼠呼吸频率与体重/肺重相比较,差异无统计学意义(P0.05)。(4)与对照组比较,实验组新生鼠血中IL-6、IL-8、MMP-9的浓度差异有统计学意义(P0.05),并且随着注射LPS浓度的增高,新生鼠血IL-6、IL-8、MMP-9的浓度也越来越高。结论:SD大鼠孕18 d腹腔注射LPS可导致孕鼠宫内感染,且在注射LPS浓度为0.7 mg/kg时,可制作稳定的宫内感染的模型,同时当注射LPS浓度大于或等于0.7 mg/kg时可造成新生鼠肺病理损害、肺水肿,且这种损害会持续至生后7天,血中的炎症因子IL-6、IL-8、MMP-9也会增高,这对研究预测宫内感染肺部疾病有着重要价值。
[Abstract]:Objective: to establish a stable pregnant rat model of intrauterine infection by intraperitoneal injection of lipopolysaccharide (LPS), and to investigate the degree of lung injury caused by intrauterine infection by pathological sections of lung tissue, and to monitor the concentration of IL-6 and IL-8 MMP9 in blood of neonatal rats. To explore the value of the aforementioned inflammatory factors in predicting pulmonary diseases. Methods: in the intrauterine infection group, 18 pregnant rats of 18 days of gestation were injected intrauterine injection of appropriate concentration of LPSV 0.7g / kg, respectively. The control group: 3 pregnant rats on the 18th day of gestation. Intrauterine injection of sterilized physiological saline (LPS) was used to observe the number of newborn rats born naturally, each of which was dried and weighed, and then was fed with the delivery mother. The respiratory rate of the newborn rats at different times of the day was recorded. Abnormal conditions, such as mobility and cyanosis. First, the placenta after natural delivery was collected. Fresh placenta was collected by elective cesarean section, and 8 newborn rats from each litter of newborn rats were randomly selected according to the age of 1, 3 and 7 days. A total of 48 lung tissue specimens were collected at each time point in each group. Blood samples were taken from randomly selected newborn rats (blood taken from the heart or blood from the inner canthus of the eyeball. Blood samples were retained for the ELISA test; lung tissue was quickly removed after death. The placenta and lung were stained with HE to prepare the sections. The mortality rate, abortion rate and death rate of newborn rats in the experimental group and control group were compared with those in the control group. Compared with the placental and lung pathological scores of neonatal rats in different LPS groups, there was significant difference in the expression of IL-6 and IL-8 MMP-9 in fetal blood by weight / lung weight Elisa. Results the concentration of LPS was higher than that of 0.7mg/kg. Compared with the control group, the placental pathology showed that inflammatory cells began to infiltrate the chorionic amniotic membrane, and the higher the concentration of LPS was, the more the inflammatory cells were. The degree of pulmonary inflammatory infiltration caused by injection of different concentrations of LPS was also different. The inflammatory scores of different groups were significantly different from those of the control group (P 0.05), suggesting that intrauterine infection could cause pulmonary inflammation. The higher the concentration of LPS was, the more inflammatory cells were infiltrated, the more damaged the lung tissue was, and the more serious the lung injury was in neonatal rats. And the injury lasted until about 7 days after birth.) the respiratory rate and body weight / lung weight of neonatal rats with different concentrations of LPS were compared between the experimental group and the control group. Compared with the control group, the concentration of IL-6 and IL-8 MMP-9 in the blood of the experimental group was significantly different from that of the control group, and the concentration of IL-6 and IL-8 MMP-9 in the blood of the experimental group increased with the increase of the concentration of LPS. Conclusion Intraperitoneal injection of LPS on the 18th day of gestation can lead to intrauterine infection in pregnant rats, and stable intrauterine infection model can be made when the concentration of LPS is 0.7 mg/kg. At the same time, when the concentration of LPS was greater than or equal to 0.7 mg/kg, lung pathological damage and pulmonary edema could be caused in neonatal rats, and the damage would last until 7 days after birth, and the inflammatory factor IL-6, IL-8, MMP-9 in blood would also increase. This is of great value for the prediction of intrauterine pulmonary diseases.
【学位授予单位】：青岛大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：R722.1

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