中华围产医学杂志
中華圍產醫學雜誌
중화위산의학잡지
CHINESE JOURNAL OF PERINATAL MEDICINE
2013年
3期
165-169
,共5页
王晓凤%滕慧云%刘敬%杨娜%任晓暾
王曉鳳%滕慧雲%劉敬%楊娜%任曉暾
왕효봉%등혜운%류경%양나%임효돈
胎儿生长迟缓%牛磺酸%细胞凋亡%胶质细胞源性神经营养因子%半胱氨酸天冬氨酸蛋白酶3
胎兒生長遲緩%牛磺痠%細胞凋亡%膠質細胞源性神經營養因子%半胱氨痠天鼕氨痠蛋白酶3
태인생장지완%우광산%세포조망%효질세포원성신경영양인자%반광안산천동안산단백매3
Fetal growth retardation%Taurine%Apoptosis%Glial cell line-derived neurotrophic factor%Caspase 3
目的 通过建立胎儿生长受限(fetal growth restriction,FGR)大鼠模型,探讨孕鼠补充牛磺酸对FGR胎鼠脑细胞凋亡以及胶质细胞源性神经营养因子(glial cell line-derived neurotrophic factor,GDNF)和半胱氨酸蛋白酶-3(caspase 3)表达的影响. 方法 将15只Sprague-Dawley (SD)孕鼠随机分为对照组、FGR模型组(模型组)、FGR+孕鼠补充牛磺酸组(牛磺酸组),每组5只.通过低蛋白饮食法建立FGR模型,牛磺酸组自妊娠第12天开始于饲料中添加牛磺酸300 mg/(kg·d)直至自然分娩.对照组每窝随机取2只适于胎龄胎鼠,牛磺酸组与模型组每窝随机取2只FGR胎鼠,应用原位末端脱氧核苷酸转移酶标记(terminal deoxynucleotidyl transferase mediated nick end labeling,TUNEL)法检测各组胎鼠脑神经细胞凋亡情况.用免疫组织化学方法检测GDNF和caspase-3表达情况.统计学方法采用单因素方差分析、Kruskal-Wallis秩和检验、SNK检验和Tamhane's检验. 结果 (1)对照组、模型组及牛磺酸组胎鼠总数分别为65、60和59只,胎鼠平均体重为(6.36±0.44)、(4.55±0.45)和(5.11±0.67)g.模型组胎鼠均发生FGR,牛磺酸组胎鼠FGR发生率为76.3％ (45/59),成功建立FGR模型.(2)对照组胎鼠脑组织内偶见TUNEL阳性细胞；模型组明显增多,在大脑皮质、海马、白质区均有分布；牛磺酸组较模型组则明显减少.3组TUNEL阳性细胞数分别为(0.46±0.11)、(14.76±3.42)和(6.78±1.93)个(H＝429.80,P＝0.000).(3)对照组胎鼠大脑皮质内仅有少量GDNF阳性细胞分布,模型组明显增多,牛磺酸组则进一步增多.3组GDNF阳性细胞数分别为(93.56±6.73)、(120.36±6.23)和(139.56±5.28)个(H=715.17,P＝0.000).(4)对照组胎鼠大脑皮质内仅见少数caspase 3阳性细胞分布,模型组明显增多,牛磺酸组较模型组显著减少,但仍多于对照组.3组caspase 3阳性细胞数分别为(7.50±2.31)、(151.32±24.43)和(37.28±11.21)个(F=132.54,P＝0.000). 结论 FGR胎鼠脑细胞凋亡显著增多.孕鼠补充牛磺酸可以显著减少FGR胎鼠脑细胞凋亡,其机制可能与牛磺酸能够促进GDNF表达和减少caspase-3表达有关.
目的 通過建立胎兒生長受限(fetal growth restriction,FGR)大鼠模型,探討孕鼠補充牛磺痠對FGR胎鼠腦細胞凋亡以及膠質細胞源性神經營養因子(glial cell line-derived neurotrophic factor,GDNF)和半胱氨痠蛋白酶-3(caspase 3)錶達的影響. 方法 將15隻Sprague-Dawley (SD)孕鼠隨機分為對照組、FGR模型組(模型組)、FGR+孕鼠補充牛磺痠組(牛磺痠組),每組5隻.通過低蛋白飲食法建立FGR模型,牛磺痠組自妊娠第12天開始于飼料中添加牛磺痠300 mg/(kg·d)直至自然分娩.對照組每窩隨機取2隻適于胎齡胎鼠,牛磺痠組與模型組每窩隨機取2隻FGR胎鼠,應用原位末耑脫氧覈苷痠轉移酶標記(terminal deoxynucleotidyl transferase mediated nick end labeling,TUNEL)法檢測各組胎鼠腦神經細胞凋亡情況.用免疫組織化學方法檢測GDNF和caspase-3錶達情況.統計學方法採用單因素方差分析、Kruskal-Wallis秩和檢驗、SNK檢驗和Tamhane's檢驗. 結果 (1)對照組、模型組及牛磺痠組胎鼠總數分彆為65、60和59隻,胎鼠平均體重為(6.36±0.44)、(4.55±0.45)和(5.11±0.67)g.模型組胎鼠均髮生FGR,牛磺痠組胎鼠FGR髮生率為76.3％ (45/59),成功建立FGR模型.(2)對照組胎鼠腦組織內偶見TUNEL暘性細胞；模型組明顯增多,在大腦皮質、海馬、白質區均有分佈；牛磺痠組較模型組則明顯減少.3組TUNEL暘性細胞數分彆為(0.46±0.11)、(14.76±3.42)和(6.78±1.93)箇(H＝429.80,P＝0.000).(3)對照組胎鼠大腦皮質內僅有少量GDNF暘性細胞分佈,模型組明顯增多,牛磺痠組則進一步增多.3組GDNF暘性細胞數分彆為(93.56±6.73)、(120.36±6.23)和(139.56±5.28)箇(H=715.17,P＝0.000).(4)對照組胎鼠大腦皮質內僅見少數caspase 3暘性細胞分佈,模型組明顯增多,牛磺痠組較模型組顯著減少,但仍多于對照組.3組caspase 3暘性細胞數分彆為(7.50±2.31)、(151.32±24.43)和(37.28±11.21)箇(F=132.54,P＝0.000). 結論 FGR胎鼠腦細胞凋亡顯著增多.孕鼠補充牛磺痠可以顯著減少FGR胎鼠腦細胞凋亡,其機製可能與牛磺痠能夠促進GDNF錶達和減少caspase-3錶達有關.
목적 통과건립태인생장수한(fetal growth restriction,FGR)대서모형,탐토잉서보충우광산대FGR태서뇌세포조망이급효질세포원성신경영양인자(glial cell line-derived neurotrophic factor,GDNF)화반광안산단백매-3(caspase 3)표체적영향. 방법 장15지Sprague-Dawley (SD)잉서수궤분위대조조、FGR모형조(모형조)、FGR+잉서보충우광산조(우광산조),매조5지.통과저단백음식법건립FGR모형,우광산조자임신제12천개시우사료중첨가우광산300 mg/(kg·d)직지자연분면.대조조매와수궤취2지괄우태령태서,우광산조여모형조매와수궤취2지FGR태서,응용원위말단탈양핵감산전이매표기(terminal deoxynucleotidyl transferase mediated nick end labeling,TUNEL)법검측각조태서뇌신경세포조망정황.용면역조직화학방법검측GDNF화caspase-3표체정황.통계학방법채용단인소방차분석、Kruskal-Wallis질화검험、SNK검험화Tamhane's검험. 결과 (1)대조조、모형조급우광산조태서총수분별위65、60화59지,태서평균체중위(6.36±0.44)、(4.55±0.45)화(5.11±0.67)g.모형조태서균발생FGR,우광산조태서FGR발생솔위76.3％ (45/59),성공건립FGR모형.(2)대조조태서뇌조직내우견TUNEL양성세포；모형조명현증다,재대뇌피질、해마、백질구균유분포；우광산조교모형조칙명현감소.3조TUNEL양성세포수분별위(0.46±0.11)、(14.76±3.42)화(6.78±1.93)개(H＝429.80,P＝0.000).(3)대조조태서대뇌피질내부유소량GDNF양성세포분포,모형조명현증다,우광산조칙진일보증다.3조GDNF양성세포수분별위(93.56±6.73)、(120.36±6.23)화(139.56±5.28)개(H=715.17,P＝0.000).(4)대조조태서대뇌피질내부견소수caspase 3양성세포분포,모형조명현증다,우광산조교모형조현저감소,단잉다우대조조.3조caspase 3양성세포수분별위(7.50±2.31)、(151.32±24.43)화(37.28±11.21)개(F=132.54,P＝0.000). 결론 FGR태서뇌세포조망현저증다.잉서보충우광산가이현저감소FGR태서뇌세포조망,기궤제가능여우광산능구촉진GDNF표체화감소caspase-3표체유관.
Objective To explore the effect of antenatal taurine supplementation on cerebral apoptosis and the expression of glial cell line-derived neurotrophic factor (GDNF) and caspase-3 in fetal rats with fetal growth restriction (FGR).Methods Fifteen pregnant Sprague-Dawley rats were randomly divided into three groups:control group,FGR model group (model group) and FGR with antenatal taurine supplementation group (taurine group).Taurine was added into the diet of taurine group at a dose of 300 mg/(kg · d) from the 12th day of gestation until natural delivery.Two appropriate for gestational age (AGA) newborn rats were randomly selected from each mother in control group and two FGR fetal rats were randomly selected from each mother both in model and taurine groups.Apoptosis of neural cells in the brain was detected by terminal deoxynucleotidyl transferase mediated nick end labeling (TUNEL).Changes in protein expression of GDNF and caspase-3 were detected by immunohistochemistry.Levene method,one-way ANOVA and SNK-test,or Kruskal Wallis rank sum test and Tamhane’ s test were applied for statistical analysis.Results (1)The total amounts of fetal rats in control group,model group and taurine group were 65,60 and 59.The mean body weight of fetal rats were (6.36±0.44) g,(4.55 ± 0.45) g and (5.11±0.67) g,respectively.All fetal rats developed FGR in model group,while 76.3％(45/59) of fetal rats were FGR in taurine group.Therefore,FGR model was successfully established.(2) In control group,there were few expression of TUNEL positive cells in cerebral cortex.A large amount of TUNEL positive cells were found in the cortex,hippocampal and white matter area in model group,but less positive cells were identified in taurine group than in model group.The amount of apoptotic brain cells in the three groups were (0.46 ± 0.11),(14.76 ± 3.42) and (6.78 ± 1.93),respectively(H=429.80,P=0.000).(3)There were only small amount of GDNF positive cells in cerebral cortex in control group and more in model group.The amount of GDNF positive cells was further increased in taurine group.The amount of GDNF positive cells in cerebral cortex in the three groups were (93.56± 6.73),(120.36± 6.23)and(139.56± 5.28),respectively (H=715.17,P=0.000).(4) Few caspase-3 positive cells were found in cerebral cortex in control group.A large amount of positive cells were found in model group and less positive cells were found in taurine group,which was still more than those in control group.The amounts of caspase-3 positive cells in the three groups were (7.50±2.31),(151.32±24.43)and(37.28±11.21),respectively (F=132.54,P=0.000).Conclusions The number of apoptotic neural cells in brain tissue of baby rats with FGR were significantly increased,which can be significantly alleviated by maternal antenatal taurine supplementation through upregulation of GDNF and downregulation of caspase-3 expression.
