中华实验外科杂志
中華實驗外科雜誌
중화실험외과잡지
CHINESE JOURNAL OF EXPERIMENTAL SURGERY
2011年
2期
264-266
,共3页
潘长福%汪泱%沈晓黎%娄远蕾%阮琼芳%邓志锋
潘長福%汪泱%瀋曉黎%婁遠蕾%阮瓊芳%鄧誌鋒
반장복%왕앙%침효려%루원뢰%원경방%산지봉
颅脑损伤%海马%神经干细胞%增殖
顱腦損傷%海馬%神經榦細胞%增殖
로뇌손상%해마%신경간세포%증식
Traumatic brain injury%Hippocampus%Neural stem cells%Proliferation
目的 观察颅脑损伤后海马区notch1相关微小RNA(miRNA)、notch1及nestin的表达变化,探讨颅脑损伤后神经干细胞增殖机制.方法 采用自由落体法复制闭合性颅脑损伤小鼠模型,于伤后4 h、1 d和3 d处死动物.(1)实时聚合酶链反应(Real-time PCR)检测伤后4 h、1 d和3d伤侧海马区mir-326、mir-34a和notch1 mRNA的表达变化.(2)免疫荧光染色检测伤后3 d伤侧海马齿状回区notch1和nestin蛋白表达变化.结果 (1)颅脑损伤后4 h、1 d和3 d组海马区mir-326和mir-34a表达水平分别为假手术组的(0.36±0.16)、(0.16±0.04)、(0.36±0.17)倍和(0.48±0.15)、(0.50±0.04)、(0.25±0.14)倍,均低于假手术组(P＜0.01);(2)颅脑损伤后4 h、1 d和3 d组海马区notch1 mRNA表达水平分别为假手术组的(1.77±0.17)、(2.55±0.48)和(2.44±0.58)倍,均高于假手术组(P＜0.05);颅脑损伤后3 d组海马齿状回区notch1阳性细胞明显多于假手术组[(18.20±3.56)个比(0.40±0.55)个,P＜0.01].(3)颅脑损伤后3 d组海马齿状回区nestin阳性细胞数明显高于假手术组[(21.80±5.07)个比(0.80±0.45)个,P＜0.01].结论 在颅脑损伤后海马区神经干细胞增殖过程中,mir-326和mir-34a表达明显降低,其靶基因notch1 mRNA和蛋白表达明显升高,提示mir-326和mir-34a可靶向抑制notch信号分子,调控伤后神经干细胞增殖过程.
目的 觀察顱腦損傷後海馬區notch1相關微小RNA(miRNA)、notch1及nestin的錶達變化,探討顱腦損傷後神經榦細胞增殖機製.方法 採用自由落體法複製閉閤性顱腦損傷小鼠模型,于傷後4 h、1 d和3 d處死動物.(1)實時聚閤酶鏈反應(Real-time PCR)檢測傷後4 h、1 d和3d傷側海馬區mir-326、mir-34a和notch1 mRNA的錶達變化.(2)免疫熒光染色檢測傷後3 d傷側海馬齒狀迴區notch1和nestin蛋白錶達變化.結果 (1)顱腦損傷後4 h、1 d和3 d組海馬區mir-326和mir-34a錶達水平分彆為假手術組的(0.36±0.16)、(0.16±0.04)、(0.36±0.17)倍和(0.48±0.15)、(0.50±0.04)、(0.25±0.14)倍,均低于假手術組(P＜0.01);(2)顱腦損傷後4 h、1 d和3 d組海馬區notch1 mRNA錶達水平分彆為假手術組的(1.77±0.17)、(2.55±0.48)和(2.44±0.58)倍,均高于假手術組(P＜0.05);顱腦損傷後3 d組海馬齒狀迴區notch1暘性細胞明顯多于假手術組[(18.20±3.56)箇比(0.40±0.55)箇,P＜0.01].(3)顱腦損傷後3 d組海馬齒狀迴區nestin暘性細胞數明顯高于假手術組[(21.80±5.07)箇比(0.80±0.45)箇,P＜0.01].結論 在顱腦損傷後海馬區神經榦細胞增殖過程中,mir-326和mir-34a錶達明顯降低,其靶基因notch1 mRNA和蛋白錶達明顯升高,提示mir-326和mir-34a可靶嚮抑製notch信號分子,調控傷後神經榦細胞增殖過程.
목적 관찰로뇌손상후해마구notch1상관미소RNA(miRNA)、notch1급nestin적표체변화,탐토로뇌손상후신경간세포증식궤제.방법 채용자유락체법복제폐합성로뇌손상소서모형,우상후4 h、1 d화3 d처사동물.(1)실시취합매련반응(Real-time PCR)검측상후4 h、1 d화3d상측해마구mir-326、mir-34a화notch1 mRNA적표체변화.(2)면역형광염색검측상후3 d상측해마치상회구notch1화nestin단백표체변화.결과 (1)로뇌손상후4 h、1 d화3 d조해마구mir-326화mir-34a표체수평분별위가수술조적(0.36±0.16)、(0.16±0.04)、(0.36±0.17)배화(0.48±0.15)、(0.50±0.04)、(0.25±0.14)배,균저우가수술조(P＜0.01);(2)로뇌손상후4 h、1 d화3 d조해마구notch1 mRNA표체수평분별위가수술조적(1.77±0.17)、(2.55±0.48)화(2.44±0.58)배,균고우가수술조(P＜0.05);로뇌손상후3 d조해마치상회구notch1양성세포명현다우가수술조[(18.20±3.56)개비(0.40±0.55)개,P＜0.01].(3)로뇌손상후3 d조해마치상회구nestin양성세포수명현고우가수술조[(21.80±5.07)개비(0.80±0.45)개,P＜0.01].결론 재로뇌손상후해마구신경간세포증식과정중,mir-326화mir-34a표체명현강저,기파기인notch1 mRNA화단백표체명현승고,제시mir-326화mir-34a가파향억제notch신호분자,조공상후신경간세포증식과정.
Objective To observe the expression of notch1-related miRNA, notch1 and nestin,and explore the regulatory mechanism of neural stem cells proliferation in hippocampus of mice after traumatic brain injury (TBI). Methods Mice were suffered closed head injury, and then sacrificed at 4th h,1st day and 3rd day post-injury. Real-time polymerase chain reaction (PCR) was used to detect the expression levels of mir-326, mir-34a and notch1 mRNA in mice hippocampus at 4th h, 1st day and 3rd day post TBI. Immunofluorescence was conducted to determine protein expression levels of notch1 and nestin in mice hippocampus at 3rd day post TBI. Results ( 1 ) Relative to sham group, the levels of mir-326 and mir-34a at 4th h, 1st day and 3rd day after TBI in the hippocampus were respectively (0. 36 ± 0. 16),(0. 16±0.04), (0.36±0. 17) and (0.48±0. 15), (0.50±0.04), (0.25 ±0. 14) fold (P＜0.01);(2) Relative to sham group, the levels of notch1 mRNA at 4th h, 1st day and 3rd day post TBI injury in the hippocampus were respectively (1.77 ±0. 17), (2.55 ±0.48) and (2.44±0.58) fold (P＜0.05).Notch1 + cells in the DG at 3rd day post TBI were significantly increased compared to sham group ( 18.20± 3.56 vs 0. 40 ±0. 55 ,P ＜0. 01 ); (3) Nestin + cells in the DG at 3rd day post TBI were increased apparently compared to sham group (21.80 ± 5. 07 vs 0.80 ± 0. 45, P ＜ 0. 01 ). Conclusion mir-326 and mir-34a may play a key role in trauma-induced neural stem cells proliferation in hippocampus in vivo by targeting notch1 signaling.
