新生7日龄高胆红素血症大鼠海马区神经元Tau、p-Tau蛋白表达及其细胞凋亡的实验研究  

Experimental Study on Expressions of Tau,p-Tau and the Apoptosis of Neuron Cells in Hippocampal Region of 7-Day Neonatal Rats with Hyperbilirubinemia

作　　者：史瑞玲 阴怀清[2] SHI Ruiling;YIN Huaiqing(First Clinical Medical College of Shanxi Medical University,Taiyuan 030001,Shanxi,China)

机构地区：[1]山西医科大学第一临床医学院,太原030001 [2]山西医科大学第一医院,太原030001

出　　处：《中西医结合心脑血管病杂志》2020年第9期1386-1391,共6页Chinese Journal of Integrative Medicine on Cardio-Cerebrovascular Disease

基　　金：山西省回国留学人员科研资助项目(No.2017-113)。

摘　　要：目的观察新生7日龄高胆红素血症大鼠海马区神经元Tau、p-Tau蛋白表达与细胞凋亡情况,探讨Tau、p-Tau蛋白在新生儿高胆红素血症所致脑损伤中的早期诊断价值。方法采用腹腔注射胆红素溶液造模。选取7日龄SD大鼠144只,随机分为3组,对照组(C组)腹腔注射等体积生理盐水0.025 mL/g;T1组腹腔注射胆红素溶液,每只100μg/g(胆红素浓度为4 mg/mL);T2组腹腔注射胆红素溶液,每只150μg/g(胆红素浓度为6 mg/mL),根据造模后处死时间不同(2 h、6 h、12 h、24 h、48 h和72 h)每组分为6个亚组,各8只。观察不同时间点新生大鼠异常行为,断头取脑切片,苏木素-伊红(HE)染色法观察海马区的病理改变,脱氧核糖核酸末端转移酶介导的缺口末端标记法(TUNEL)检测细胞凋亡,免疫组化法检测Tau、p-Tau蛋白表达。结果时间对海马区神经元Tau、p-Tau蛋白表达及神经元细胞凋亡率总变异的单效应影响依次为36.79%(F=198.7,P<0.05),33.99%(F=183.2,P<0.05),45.97%(F=616.8,P<0.05);胆红素剂量对Tau、p-Tau蛋白表达及神经元细胞凋亡率总变异的单效应影响分别为41.81%(F=564.5,P<0.05),45.23%(F=609.5,P<0.05),32.48%(F=1090.0,P<0.05);且时间与胆红素剂量对上述观察指标存在交互作用。C组各时间点神经元Tau、p-Tau蛋白表达及神经元细胞凋亡率总变异表达无明显变化。造模后6 h、12 h、24 h,T1组、T2组神经元Tau、p-Tau蛋白与细胞凋亡率存在线性正相关(r=0.9677,P<0.05;r=0.5928,P<0.05;r=0.5242,P<0.05)。结论过表达Tau、p-Tau蛋白参与早期新生儿高胆红素血症所致脑损伤的发生发展,Tau、p-Tau蛋白可作为早期诊断和治疗新生儿胆红素脑病的生化参考指标之一。Objective To explore the possible mechanism of neonatal hyperbilirubinemia brain injury through the relationship between Tau,p-Tau protein expression,and hippocampal neuron cells apoptosis in seven-day-old rats of hyperbilirubinemia model,so as to provide experimental basis for its early clinical diagnosis and treatment.Methods The model was made by intraperitoneal injection of bilirubin solution.According to different intervention dose and killing time,a total of 144 SD rats(7 days)were selected and randomly divided into three groups:control group(Group C,0.025 mL/g saline through intraperitoneal injection),and test group(Group T1,100μg/g bilirubin solution through intraperitoneal injection,bilirubin concentration 4 mg/mL;Group T2,150μg/g bilirubin solution through intraperitoneal injection,bilirubin concentration 6 mg/mL),each group with six subgroups,with eight rats in each subgroups.Abnormal behaviors at different time points(2 h、6 h、12 h、24 h、48 h、72 h after molding)were observed.Brain tissue sections were made.Then,pathological changes in hippocampus were observed by HE staining,apoptosis was detected by TUNEL assay,Tau and p-Tau expressions were measured by immunohistochemistry.Finally,the Grahphad Prism 8.0 software was used for multivariate variance analysis and correlation analysis.Results Time as simple effect on total variations of Tau protein,p-Tau protein expression,and neuronal apoptosis in hippocampal neurons was 36.79%(F=198.7,P<0.05),33.99%(F=183.2,P<0.05),and 45.97%(F=616.8,P<0.05)respectively.Bilirubin dose as simple effects on above three indexes’total variations was 41.81%(F=564.5,P<0.05),45.23%(F=609.5,P<0.05),and 32.48%(F=1090.0,P<0.05),respectively.In addition,time and bilirubin dose had an interactive effect on the above indexes.There were no significant changes in expressions of above three indicators at different times in group C.At 6 h,12 h,and 24 h,Tau,p-Tau protein,and neuronal apoptosis rate showed a positive linear correlation(r=0.9677,P<0.05;r=0.5928,P<0.05;r=0.5242

关 键 词：高胆红素血症 海马 TAU蛋白 p-Tau蛋白 神经元细胞凋亡 实验研究 

分 类 号：R722.1[医药卫生—儿科] R259[医药卫生—临床医学]