中华儿科杂志
中華兒科雜誌
중화인과잡지
Chinese Journal of Pediatrics
2012年
5期
361-365
,共5页
唐小曼%覃怡%廖淳杰%谢玉波%蓝雨雁
唐小曼%覃怡%廖淳傑%謝玉波%藍雨雁
당소만%담이%료순걸%사옥파%람우안
异丙酚%海马%半胱氨酸天冬氨酸蛋白酶3%生存素
異丙酚%海馬%半胱氨痠天鼕氨痠蛋白酶3%生存素
이병분%해마%반광안산천동안산단백매3%생존소
Propofol%Hippocampus%Caspases 3%Survivin
目的 观察异丙酚麻醉对新生大鼠海马神经元形态结构、生存素( Survivin)和半胱氨酸天冬氨酸蛋白酶3( Caspase-3)基因表达的影响.方法 将体重10～15 g的7日龄雄性SID大鼠100只,以数字随机法随机分为4组,每组25只.对照组(A组)不注射任何药物,其他3组分别腹腔注射异丙酚50 mg/kg(R组)、异丙酚100 mg/kg(C组)或异丙酚200 mg/kg(D组).血气分析仪检测大鼠动脉血呼吸和代谢指标的变化,透射电镜观察海马CA1区神经元细胞形态学变化,Fluoro-Jade B (FJB)荧光染色法检测海马CA1区变性、坏死、凋亡的神经元,半定量逆转录-聚合酶链反应(RTPCR)和Western blot技术检测海马组织中Survivin和Caspase-3基因mRNA和蛋白的表达.结果 麻醉维持时间:D组＞C组＞B组；各组犬鼠动脉血pH、氧分压(PaO2)、二氧化碳分压(PaCO2)、HCO3-、碱剩余(BE)、氧饱和度( SaO2)之间比较差异无统计学意义(P＞0.05).A组、B组海马神经元结构基本正常,C组神经元细胞核肿胀,D组核碎裂、染色质边集甚至出现凋亡小体；A组、B组、C组、D组大鼠海马 CA1区FJB阳性细胞数分别为(0.6±0.3)、(2.5±1.3)、(7.1±2.3)、(9.4±2.6),B组、C组、D组与A组比较差异有统计学意义(p＜0.05),而且FJB阳性细胞数:D组＞C组＞B组.A组、B组、C组海马组织Caspase-3 mRNA的表达量分别为(0.78±0.12)、(0.84±0.17)、(0.89±0.19),Caspase-3蛋白的表达量分别为(0.22±0.05) 、(0.26±0.07)、(0.21±0.06)；A组、B组、C组Survivin mRNA的表达量分别为(0.56±0.12)、(0.58±0.15)、(0.53±0.16),Survivin蛋白的表达量分别为(0.24±0.07)、(0.21±0.05)、(0.23±0.06)；A组、B组、C组Caspase-3和Survivin mRNA和蛋白的表达差异无统计学意义(P＞0.05)；然而,D组海马组织Caspase-3 mRNA和蛋白的表达量分别为(1.21±0.14)、(0.42±0.12),高于A组、B组、C组(P＜0.05)；D组海马组织Survivin mRNA和蛋白的表达量分别为(0.41±0.11)、(0.12±0.03),低于A组、B组、C组(p＜0.05).结论 较大剂量异内酚可以通过非麻醉缺氧作用促进海马神经元细胞的变性、坏死和凋亡,使海马组织Caspase-3的活性增强而Survivin的表达减少.
目的 觀察異丙酚痳醉對新生大鼠海馬神經元形態結構、生存素( Survivin)和半胱氨痠天鼕氨痠蛋白酶3( Caspase-3)基因錶達的影響.方法 將體重10～15 g的7日齡雄性SID大鼠100隻,以數字隨機法隨機分為4組,每組25隻.對照組(A組)不註射任何藥物,其他3組分彆腹腔註射異丙酚50 mg/kg(R組)、異丙酚100 mg/kg(C組)或異丙酚200 mg/kg(D組).血氣分析儀檢測大鼠動脈血呼吸和代謝指標的變化,透射電鏡觀察海馬CA1區神經元細胞形態學變化,Fluoro-Jade B (FJB)熒光染色法檢測海馬CA1區變性、壞死、凋亡的神經元,半定量逆轉錄-聚閤酶鏈反應(RTPCR)和Western blot技術檢測海馬組織中Survivin和Caspase-3基因mRNA和蛋白的錶達.結果 痳醉維持時間:D組＞C組＞B組；各組犬鼠動脈血pH、氧分壓(PaO2)、二氧化碳分壓(PaCO2)、HCO3-、堿剩餘(BE)、氧飽和度( SaO2)之間比較差異無統計學意義(P＞0.05).A組、B組海馬神經元結構基本正常,C組神經元細胞覈腫脹,D組覈碎裂、染色質邊集甚至齣現凋亡小體；A組、B組、C組、D組大鼠海馬 CA1區FJB暘性細胞數分彆為(0.6±0.3)、(2.5±1.3)、(7.1±2.3)、(9.4±2.6),B組、C組、D組與A組比較差異有統計學意義(p＜0.05),而且FJB暘性細胞數:D組＞C組＞B組.A組、B組、C組海馬組織Caspase-3 mRNA的錶達量分彆為(0.78±0.12)、(0.84±0.17)、(0.89±0.19),Caspase-3蛋白的錶達量分彆為(0.22±0.05) 、(0.26±0.07)、(0.21±0.06)；A組、B組、C組Survivin mRNA的錶達量分彆為(0.56±0.12)、(0.58±0.15)、(0.53±0.16),Survivin蛋白的錶達量分彆為(0.24±0.07)、(0.21±0.05)、(0.23±0.06)；A組、B組、C組Caspase-3和Survivin mRNA和蛋白的錶達差異無統計學意義(P＞0.05)；然而,D組海馬組織Caspase-3 mRNA和蛋白的錶達量分彆為(1.21±0.14)、(0.42±0.12),高于A組、B組、C組(P＜0.05)；D組海馬組織Survivin mRNA和蛋白的錶達量分彆為(0.41±0.11)、(0.12±0.03),低于A組、B組、C組(p＜0.05).結論 較大劑量異內酚可以通過非痳醉缺氧作用促進海馬神經元細胞的變性、壞死和凋亡,使海馬組織Caspase-3的活性增彊而Survivin的錶達減少.
목적 관찰이병분마취대신생대서해마신경원형태결구、생존소( Survivin)화반광안산천동안산단백매3( Caspase-3)기인표체적영향.방법 장체중10～15 g적7일령웅성SID대서100지,이수자수궤법수궤분위4조,매조25지.대조조(A조)불주사임하약물,기타3조분별복강주사이병분50 mg/kg(R조)、이병분100 mg/kg(C조)혹이병분200 mg/kg(D조).혈기분석의검측대서동맥혈호흡화대사지표적변화,투사전경관찰해마CA1구신경원세포형태학변화,Fluoro-Jade B (FJB)형광염색법검측해마CA1구변성、배사、조망적신경원,반정량역전록-취합매련반응(RTPCR)화Western blot기술검측해마조직중Survivin화Caspase-3기인mRNA화단백적표체.결과 마취유지시간:D조＞C조＞B조；각조견서동맥혈pH、양분압(PaO2)、이양화탄분압(PaCO2)、HCO3-、감잉여(BE)、양포화도( SaO2)지간비교차이무통계학의의(P＞0.05).A조、B조해마신경원결구기본정상,C조신경원세포핵종창,D조핵쇄렬、염색질변집심지출현조망소체；A조、B조、C조、D조대서해마 CA1구FJB양성세포수분별위(0.6±0.3)、(2.5±1.3)、(7.1±2.3)、(9.4±2.6),B조、C조、D조여A조비교차이유통계학의의(p＜0.05),이차FJB양성세포수:D조＞C조＞B조.A조、B조、C조해마조직Caspase-3 mRNA적표체량분별위(0.78±0.12)、(0.84±0.17)、(0.89±0.19),Caspase-3단백적표체량분별위(0.22±0.05) 、(0.26±0.07)、(0.21±0.06)；A조、B조、C조Survivin mRNA적표체량분별위(0.56±0.12)、(0.58±0.15)、(0.53±0.16),Survivin단백적표체량분별위(0.24±0.07)、(0.21±0.05)、(0.23±0.06)；A조、B조、C조Caspase-3화Survivin mRNA화단백적표체차이무통계학의의(P＞0.05)；연이,D조해마조직Caspase-3 mRNA화단백적표체량분별위(1.21±0.14)、(0.42±0.12),고우A조、B조、C조(P＜0.05)；D조해마조직Survivin mRNA화단백적표체량분별위(0.41±0.11)、(0.12±0.03),저우A조、B조、C조(p＜0.05).결론 교대제량이내분가이통과비마취결양작용촉진해마신경원세포적변성、배사화조망,사해마조직Caspase-3적활성증강이Survivin적표체감소.
Objective Intravenous anesthetics,such as propofol,are widely used in general anesthesia.Neurodegeneration and neurocognitive impairment after exposure to propofol in neonatal rats have raised concerns regarding the safety of pediatric anesthesia.We examined the effects of neonatal propofol exposure on brain cell viability,as well as expression of hippocampal survivin and Caspase-3 mRNA and protein.Methods One hundred male Sprague-Dawley rats aged 7 d that were weighed 10-15 g were randomly divided into 4 groups (n =25 each group).Group A:the rats were injected with no drugs.Group B:the rats were intraperitoneally injected with 50 mg/kg propofol. Group C: the rats were first intraperitoneally injected with 50 mg/kg propofol and another 50 mg/kg propofol was used when the dynamic response of rats appeared again. Group D:the rats were first intraperitoneally injected with 50 mg/kg propofol and another 50 mg/kg propofol was used three times once the dynamic response of rats appeared.To study the effects of propofol exposure on respiratory and metabolic function,arterial blood was aspirated from the left ventricle of neonatal rats 2 h after discontinuation of propofol.pH,PaO2,PaCO2,HCO3-,BE and SaO2 were detected by blood gas analyzer.Moreover,to examine the effects of propofol exposure on shortterm cellular viability,the ultrastructure of neurons was observed by transmission electron microscope and Fluoro-Jade B (FJB) staining was performed to examine neuronal degeneration in hippocampal CA1 region of neonatal rats.Survivin and Caspase-3 mRNA and protein expression in hippocampus were detected by semiquantitative reverse transcription polymerase chain reaction ( RT-PCR ) and Western blotting 2 h after discontinuation of propofol.Results The time of anesthesia maintaince in newborn rats was the longest in Group D and the time of anesthesia maintaince in Group C was longer than that in Group B.Two hours after discontinuation of propofol,pH,PaO2,PaCO2,HCO3-,BE and SaO2 of arterial blood in rats were not significantly different among groups A,B,C and D ( P ＞ 0.05 ).The structure of hippocampal neurons was normal in Group A and Group B while 100 mg/kg propofol resulted in nuclear blebbing and 200 mg/kg propofol led to nuclear fragmentation,chromatin condensation and apoptotic bodies.Cellular degeneration,as measured by Fluoro-Jade B staining,siguificantly increased in hippocampal CA1 region in the anesthesia groups compared with littermates in the no anesthesia group.FJB-positive stained degenerative neurons in groups B,C and D were ( 2.5 ± 1.3 ),( 7.1 ± 2.3 ) and ( 9.4 ± 2.6 ),which were different from that in Group A (0.6 ± 0.3 ) ( P ＜ 0.05 ).Moreover,the number of FJB-positive neurons was the highest in Group D,that in Group C was more than that in Group B.At the same time point,apoptosis was measured by expression of Caspase-3 and Survivin mRNA and protein in hippocampus of rats.Caspase-3 mRNA in groups A,B and C was (0.78 ±0.12),(0.84 ±0.17) and (0.89 ±0.19),while Caspase-3 protein in groups A,B and C was (0.22 ±0.05),(0.26±0.07) and (0.21 ±0.06).Survivin mRNA in groups A,B and C was (0.56 ± 0.12 ),(0.58 ± 0.15 ) and (0.53 ± 0.16 ),while Survivin protein in these 3 groups was (0.24±0.07),(0.21 ±0.05) and (0.23 ±0.06).Compared with that in Group A,Caspase-3 and Survivin mRNA and protein were not significantly different among Group B and Group C ( P ＞ 0.05 ).However,Caspase-3 mRNA and protein in Group D were (1.21 ±0.14) and (0.42 ±0.12),which were higher than that in the other 3 groups( P ＜0.05 ).Survivin mRNA and protein in Group D were lower than that in the other 3 groups ( P ＜ 0.05 ).Conclusions A high dose of propofol exposure may destroy the structure of neurons,induce neurodegeneration,increase Caspase-3 activity and inhibit survivin expression in hippocampus of newborn rats in vivo.
