本文選題：宮內感染　切入點：脂多糖　出處：《青島大學》2017年碩士論文

【摘要】：目的:采用腹腔注射脂多糖(lipopolysaccharide,LPS)的方法構建穩(wěn)定的宮內感染的孕鼠模型,通過肺組織病理切片了解宮內感染對肺部組織造成的損傷程度,監(jiān)測新生鼠血中IL-6、IL-8、MMP9的濃度,探究前述炎癥因子對預測肺部疾病的價值。方法:宮內感染組:孕18天的孕鼠18只,分別于宮腔內注射合適濃度的LPS(0.6、0.7、0.8、0.9、1.0mg/kg)。對照組:孕18天的孕鼠3只,于宮腔內注射和感染組LPS等量的滅菌生理鹽水。查看自然分娩的新生大鼠存活數(shù)量,每只均擦干稱重,再與分娩母鼠共同飼養(yǎng),記錄每天不同時刻新生大鼠的呼吸頻率、動度以及有無發(fā)紺等異常情況;首先收集自然分娩后胎盤(自然分娩下收集胎盤容易失敗,可選擇擇期剖宮產收集新鮮胎盤);按照1、3、7日齡分別隨機選取兩組中自然分娩所得的每窩新生大鼠中的8只新生大鼠,每組每個時間點共48份肺組織標本。將隨機選取的新生大鼠斷頭取血(心臟取血或眼球內眥取血),留取血標本用于ELISA實驗;處死后迅速取出肺組織,將所取的胎盤與肺均進行HE染色制成切片備用。比較實驗組與對照組中隨著藥物劑量遞減孕鼠死亡率、流產率以及新生大鼠死亡率的變化,比較不同LPS組新生鼠胎盤及肺病理評分、新生鼠體重/肺重、ELISA法監(jiān)測胎鼠血中的IL-6、IL-8、MMP-9表達差異有無統(tǒng)計學意義。結果:(1)注射LPS濃度大于0.7mg/kg時,各組胎盤病理與對照組相比較差異有統(tǒng)計學意義(P0.05),胎盤病理可見存在炎癥細胞開始浸潤絨毛膜羊膜,且注射孕鼠LPS濃度越高,炎癥細胞愈多,浸潤組織部位越多。(2)通過注射不同濃度LPS造成肺部炎癥浸潤的程度也不相同,不同組炎癥評分與對照組比較差異均有統(tǒng)計學意義(P0.05),提示宮內感染可造成肺部炎癥,當注射LPS濃度越高,肺部組織炎癥細胞浸潤越多,肺部組織破壞越多,新生大鼠肺部損傷越重,并且這種損傷會持續(xù)至生后7天左右。(3)注射不同濃度LPS的新生大鼠每天呼吸情況及生長指標相比較,實驗組與對照組新生大鼠呼吸頻率與體重/肺重相比較,差異無統(tǒng)計學意義(P0.05)。(4)與對照組比較,實驗組新生鼠血中IL-6、IL-8、MMP-9的濃度差異有統(tǒng)計學意義(P0.05),并且隨著注射LPS濃度的增高,新生鼠血IL-6、IL-8、MMP-9的濃度也越來越高。結論:SD大鼠孕18 d腹腔注射LPS可導致孕鼠宮內感染,且在注射LPS濃度為0.7 mg/kg時,可制作穩(wěn)定的宮內感染的模型,同時當注射LPS濃度大于或等于0.7 mg/kg時可造成新生鼠肺病理損害、肺水腫,且這種損害會持續(xù)至生后7天,血中的炎癥因子IL-6、IL-8、MMP-9也會增高,這對研究預測宮內感染肺部疾病有著重要價值。
[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.
【學位授予單位】：青島大學
【學位級別】：碩士
【學位授予年份】：2017
【分類號】：R722.1

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