摘要
目的探讨胆道闭锁(biliary atresia,BA)小鼠生存晚期的脑损伤情况及N-乙酰半胱氨酸(N-acetylcysteine,NAC)对BA小鼠脑损伤的治疗作用。方法将160只新生BALB/c小鼠按随机数字表法平均分为疾病组、空白对照组、治疗组及药物对照组,每组各40只。疾病组出生后第3天腹腔注射30μl恒河猴轮状病毒(rhesus rotavirus,RRV)诱导BA小鼠模型;治疗组出生后第3天注射等量RRV后每日腹腔注射10 mg/kg的NAC治疗至第12天;空白对照组出生后第3天予腹腔注射30μl未扩增RRV的MA104细胞培养上清;药物对照组出生后第3天起每日腹腔注射10 mg/kg的NAC至第12天。取第19天小鼠进行神经功能评分,检测血浆脑损伤标志物神经元特异性烯醇化酶(neuron specific enolase,NSE)和中枢神经特异蛋白S100β水平。评估小鼠海马锥体细胞层病理切片苏木精-伊红(hematoxylin and eosin,HE)染色、尼氏染色及Iba1免疫组织化学染色情况。评估小鼠肝门管区HE染色、细胞角蛋白19(cytokeratin 19,CK19)免疫组织化学染色及天狼猩红染色情况。组间样本神经功能评分、血浆脑损伤标志物水平比较采用t检验,组间样本海马锥体细胞层HE染色评分、尼氏染色评分、Iba1免疫组织化学染色评分比较采用Wilcoxon秩和检验。结果生后第19天疾病组小鼠神经功能评分中一般评分及局灶评分分别为(10.24±1.27)分、(6.84±1.05)分,与空白对照组小鼠均为0分比较,一般评分(t=-39.90,P<0.001)、局灶评分(t=-32.03,P<0.001)差异均有统计学意义。出生后第19天治疗组小鼠神经功能评分中一般评分及局灶评分分别为(4.84±0.92)分、(1.88±0.59)分,与疾病组小鼠比较,一般评分(t=16.81,P<0.001)、局灶评分(t=20.25,P<0.001)差异均有统计学意义。出生后第19天药物对照组小鼠神经功能评分一般评分及局灶评分均为0分。疾病组与空白对照组小鼠血浆NSE水平分别为(27.79±2.50)ng/ml、(18.93±2.78)ng/ml,差异有�
Objective To explore the status of brain injury in late survival mice with biliary atresia(BA)and to examine the therapeutic efficacy of N-acetylcysteine(NAC)for brain injury in BA mice.Methods A total of 160 neonatal Balb/c mice were randomized into four groups of blank control,disease,treatment and drug control(n=40 each).Treatment protocols of each group:Disease group adopted the BA murine model of chronic liver injury induced by an intraperitoneal injection of 30μl rhesus rotavirus(RRV)at Day 3 post-birth;Treatment group received a daily intraperitoneal injection of 10 mg/kg of NAC after an injection of 30μl RRV until Day 12 post-birth.Blank control group received 30μl MA104 cell culture supernatant without RRV amplification at Day 3 post-birth.Drug control group received a daily intraperitoneal injection of 10 mg/kg NAC from Day 3-12 post-birth.Twenty-five mice at Day 19 in each group were selected for neurological function score.Plasma levels of neuron-specific enolase(NSE)and central nervous system specific protein(S100β),markers of brain injury,were assayed in 21 subjects from each group.Hematoxylin and eosin(HE)and Nissl stains of pathological sections were evaluated with 5 mice in each group and 640×visual fields were randomly selected for each mouse.Three mice in each group were selected and 540×visual fields in each mouse were randomized for evaluating ionized calcium binding adapter molecule 1(Iba1)in pathological sections.HE,cytokeratin 19(CK19)immunohistochemical and Sirius scarlet stains were evaluated in 3 mice from each group.Student t-test was employed for comparing neurological function scores and plasma markers of brain injury between two groups.Wilcoxon rank-sum test was utilized for comparing HE scores,Nissl and Iba1 immunohistochemical stain scores in hippocampal pyramidal cell layer between two groups.Results At Day 19 post-birth,general and focal scores of disease group were(10.24±1.27)and(6.84±1.05)points respectively.As compared with blank control group with 0 points,the differ
作者
赵子阳
肖依依
马思格
童燕陆
陈虹交
王贺珍
陈严
张锐忠
夏慧敏
Zhao Ziyang;Xiao Yiyi;Ma Sige;Tong Yanlu;Chen Hongjiao;Wang Hezhen;Chen Yan;Zhang Ruizhong;Xia Huimin(First School of Clinical Medicine,Southern Medical University,Guangzhou 510515,China;Guangdong Provincial Key Laboratory of Research in Structural Birth Defect Disease,Guangzhou Women&Children's Medical Center,Guangzhou 510620,China)
出处
《中华小儿外科杂志》
CSCD
北大核心
2023年第7期648-655,共8页
Chinese Journal of Pediatric Surgery
基金
国家自然科学基金(81974056)
科技部重大项目(2021ZD0200522)
广州市科技计划项目(202206080002)。
