CAJ | 학술논문

目的探讨重组腺相关病毒载体(recombinant adeno-associated virus,rAAV)介导的脑源性神经营养因子(BDNF)对先天性脊柱裂中神经细胞凋亡的抑制作用.方法利用维甲酸灌胃的方法对18只Wistar孕鼠致畸制作先天性脊柱裂胎鼠动物模型,其中BDNF转基因治疗组和空载体治疗组各9例,于孕16 d(E16)进行胎仔外科手术,分别向显性脊柱裂胎鼠腰骶部注射1μl(大约2×105 TU) rAAV-BDNF和rAAV-ZsGreen病毒悬液；正常对照组孕鼠10例,不做任何处理.结果转基因治疗组胎鼠的存活率为70.0％,空载体治疗组胎鼠的存活率为71.4％,两组比较差异无统计掣意义(P＞0.05).BDNF转基因治疗组BDNF的表达明显高于空载体治疗组和正常组.TUNEL染色显示BDNF转基因治疗组平均每个脊髓切片的凋亡细胞数较空载体治疗组和正常对照组明显减少,各组之间的差别有统计学意义(P＜0.05).结论 BDNF转基因疗法能够显著抑制先天性脊柱裂胎鼠脊髓缺损组织神经细胞的凋亡,对显性脊柱裂治疗具有潜在应用价值.
목적 탐토중조선상관병독재체(recombinant adeno-associated virus,rAAV)개도적뇌원성신경영양인자(BDNF)대선천성척주렬중신경세포조망적억제작용.방법 이용유갑산관위적방법대18지Wistar잉서치기제작선천성척주렬태서동물모형,기중BDNF전기인치료조화공재체치료조각9례,우잉16 d(E16)진행태자외과수술,분별향현성척주렬태서요저부주사1μl(대약2×105 TU) rAAV-BDNF화rAAV-ZsGreen병독현액；정상대조조잉서10례,불주임하처리.결과 전기인치료조태서적존활솔위70.0％,공재체치료조태서적존활솔위71.4％,량조비교차이무통계체의의(P＞0.05).BDNF전기인치료조BDNF적표체명현고우공재체치료조화정상조.TUNEL염색현시BDNF전기인치료조평균매개척수절편적조망세포수교공재체치료조화정상대조조명현감소,각조지간적차별유통계학의의(P＜0.05).결론 BDNF전기인요법능구현저억제선천성척주렬태서척수결손조직신경세포적조망,대현성척주렬치료구유잠재응용개치.
Objective The purpose ot the current study was to determine the inhibition of recombinant adeno-associated viral vector-mediated brain-derived neurotrophic factor (rAAV-BDNF) on apoptosis of nerve cells in congenital spina bifida.Methods Using retinoic acid gavage teratogenicity,18 pregnant Wistar rats were used in a congenital dominant spina bifida model and divided equally into 2 groups.At 16 days gestation (E16),1μ (approximately 2 × 105 of TU) of rAAV-BDNF and rAAV-ZsGreen virus suspensions were injected into the fetal rat lumbosacral region (BDNF gene therapy and empty vector treatment groups,respectively).Rats in the control group (n =10) were left intact.Resu1ts After gene therapy,the treatment group fetal survival rate was 70.00％,and the empty vector treatment group fetal survival rate was 71.43％;the fetal rat survival rates were not significantly different.The expression of BDNF in the treatment group was significantly increased compared with the empty vector treatment and control groups.TUNEL staining showed that the average number of apoptotic cells in each spinal cord slice in the treatment group was significantly decreased compared with the empty vector treatment and control groups.Conclusions The use of genetically-modified therapy to inject rAAV-BDNF into congenital spinal bifida fetal rat spinal cord defects significantly inhibited neuronal apoptosis,which has potential application to treat dominant spina bifida.