摘要
目的探究孕期血管紧张素Ⅱ1型受体自身抗体(angiotensinⅡtype 1 receptor autoantibody,AT1-AA)暴露对胎鼠肝脏发育的影响。方法将36只10周龄健康Sprague-Dawley孕鼠随机分为对照组和AT1-AA组,每组各18只,分别于孕13 d和15 d鼠尾静脉注射生理盐水0.5 ml或AT1-AA 20μg/g。孕12 d、14 d、16 d、18 d采用酶联免疫吸附法(enzyme linked immunosorbent assay,ELISA)检测各组孕鼠血清AT1-AA浓度、尾套法测量血压。于孕14 d、16 d、18 d分别处死孕鼠,从每只孕鼠取出2~3只胎鼠进行称量,计算胚胎重量、胎鼠肝脏重量和胎鼠肝脏重量/胚胎体重比值,免疫组织化学染色法检测胎鼠肝脏血浆白蛋白(Albumin)、细胞生长因子受体(c-Kit)蛋白表达,以及RNA转录组测序(RNA sequencing,RNA-seq)。体外培养HepG2细胞,分为溶剂对照组、AT1-AA处理细胞组和AT1-AA+氯沙坦处理细胞组,CCK8试剂盒检测AT1-AA对HepG2细胞活力的影响。采用独立样本t检验、单因素方差分析对数据进行统计学分析。结果(1)孕16 d、18 d,AT1-AA组孕鼠血压显著高于对照组[(116.17±9.87)与(98.50±8.75)mmHg(1 mmHg=0.133 kPa)、(125.00±9.80)与(99.43±11.76)mmHg,t值分别为3.28和4.09,P值均<0.05]。孕14 d、16 d、18 d,AT1-AA组血清AT1-AA高于对照组(1.44±0.09与0.45±0.09、2.03±0.09与0.47±0.06、2.28±0.08与0.42±0.08,t值分别为19.20、21.64和38.68,P值均<0.05)。提示AT1-AA组孕鼠造模成功。(2)孕14 d、16 d、18 d,AT1-AA组胚胎重量与同期对照组相比均明显降低[(248.27±19.63)与(263.12±16.22)mg、(626.83±167.49)与(788.96±32.25)mg、(841.43±57.57)与(966.73±33.68)mg,t值分别为2.18、3.43和6.20,P值均<0.05]。孕16 d、18 d,AT1-AA组胎鼠肝脏重量较同期对照组显著降低[(46.97±14.00)与(73.75±6.55)mg、(60.56±7.11)与(91.75±6.19)mg,t值分别6.28和10.94,P值均<0.05];AT1-AA组胎鼠肝脏重量/胚胎重量的比值较同期对照组显著降低[(7.45±0.70)%与(9.34±0.64)%、(7.14±0.84)%与(9.45±0.76)%
Objective To investigate the effects of prenatal exposure to angiotensinⅡtype 1 receptor autoantibody(AT1-AA)on fetal liver development.Methods Thirty-six healthy Sprague-Dawley pregnant rats aged 10 weeks were randomly divided into the AT1-AA group(n=18)and the control group(n=18),which were injected with AT1-AA 20μg/g or 0.5 ml of normal saline through tail vein at 13 and 15 days of gestation,respectively.At 12,14,16,and 18 days of gestation,serum AT1-AA concentrations of the two groups were detected by enzyme linked immunosorbent assay(ELISA)and blood pressure was measured by tail-cuff methods.On the 14,16,and 18 days of gestation,the pregnant mice were killed separately and two or three fetal mice of each pregnant mouse were randomly selected for weighing.The fetal weight,fetal liver weight,and fetal liver weight/fetal weight ratio were calculated and the expression of Albumin and c-Kit protein in the fetal liver was measured by immunohistochemical staining.Fetal liver tissue was collected for RNA sequencing(RNA-seq).HepG2 cells were cultured in vitro,which were further divided into the solvent control group,AT1-AA treated cell group,and AT1-AA+losartan treated cell group.The effect of AT1-AA on the viability of HepG2 cells was detected by CCK8 kit.Independent sample t-test and one-way analysis of variance were used for the statistical analysis.Results(1)On the day 16 and 18 after conceiving,the blood pressure of pregnant mice in the AT1-AA group was significantly higher than that of the control group[(116.17±9.87)vs.(98.50±8.75)mmHg(1 mmHg=0.133 kPa),(125.00±9.80)vs.(99.43±11.76)mmHg;t=3.28 and 4.09;both P<0.05].On day 14,16,and 18,the serum AT1-AA levels in the AT1-AA group were higher than those in the control group(1.44±0.09 vs.0.45±0.09,2.03±0.09 vs.0.47±0.06,2.28±0.08 vs.0.42±0.08;t=19.20,21.64,and 38.68;all P<0.05).These findings suggest that the AT1-AA group was successfully modeled.(2)On day 14,16,and 18,the fetal weights in the AT1-AA group were significantly less than those in the cont
作者
孙婧
杨自宇
王鹏丽
王彤彤
刘慧荣
张苏丽
Sun Jing;Yang Ziyu;Wang Pengli;Wang Tongtong;Liu Huirong;Zhang Suli(Department of Physiology,Faculty of Basic Medicine,Changzhi Medical College,Changzhi 046000,China;Cultivation Base of Shanxi Key Laboratory of Research and Translational Application of Aging Mechanism,Changzhi 046000,China;Beijing Key Laboratory of Metabolic Disorder Related Cardiovascular Disease,Beijing 100069,China;Department of Physiology and Pathophysiology,College of Basic Medicine,Capital Medical University,Beijing 100069,China)
出处
《中华围产医学杂志》
CAS
CSCD
北大核心
2024年第4期301-311,共11页
Chinese Journal of Perinatal Medicine
基金
细胞生理学教育部重点实验室(山西医科大学)开放基金(KLMEC/SXMU-202203)
商丘医学高等专科学校开放课题(KFKT23001)。
