CAJ | 학술논문

目的探讨脑源性神经营养因子(brain-derived neurotrophic factor,BDNF)基因转染的骨髓间充质干细胞(bone-marrow mesenchymal stem cells,BMSC)对氨基糖苷类抗生素(aminoglycoside antibiotics,AmAn)损伤的耳蜗螺旋神经节细胞(spiral ganglion cells,SGC)的保护作用.方法采用兔抗鼠巢蛋白(Nestin)、神经元特异性烯醇化酶(neurone specific enolase,NSE)、胶质纤维酸性蛋白(glial fibrillary acidic protein,GFAP)抗体免疫组化检测在体外转染了BDNF基因的BMSC(BDNF-BMSC)分化结果 .将BDNF-BMSC植入阿米卡星致聋豚鼠的耳蜗,并设单纯植入人工外淋巴组、BMSC植入组及BDNF基因植入组作为对照.分别在移植后1、2及4周取耳蜗组织,石蜡包埋切片,HE染色观察耳蜗病理改变、计算SGC密度;免疫组化染色计算相对吸光度值,比较各组对耳蜗SGC的保护作用.结果转染后BDNF-BMSC的Nestin、NSE、GFAP阳性率均高于未转染的BMSC(P值均<0.01).耳蜗注射后在相同时间点各组间SGC密度和吸光度值差异均具有统计学意义(P值均<0.05),其中BDNF-BMSC组SGC密度最大、吸光度值最大;各组SGC密度和吸光度值在术后4周时比术后1周和2周时下降(P值均<0.05).结论豚鼠耳蜗内移植BMSC、BDNF基因、BDNF-BMSC均可拮抗阿米卡星对SGC的损伤,其中BDNF-BMSC的保护作用最强.
목적 탐토뇌원성신경영양인자(brain-derived neurotrophic factor,BDNF)기인전염적골수간충질간세포(bone-marrow mesenchymal stem cells,BMSC)대안기당감류항생소(aminoglycoside antibiotics,AmAn)손상적이와라선신경절세포(spiral ganglion cells,SGC)적보호작용.방법 채용토항서소단백(Nestin)、신경원특이성희순화매(neurone specific enolase,NSE)、효질섬유산성단백(glial fibrillary acidic protein,GFAP)항체면역조화검측재체외전염료BDNF기인적BMSC(BDNF-BMSC)분화결과 .장BDNF-BMSC식입아미잡성치롱돈서적이와,병설단순식입인공외림파조、BMSC식입조급BDNF기인식입조작위대조.분별재이식후1、2급4주취이와조직,석사포매절편,HE염색관찰이와병리개변、계산SGC밀도;면역조화염색계산상대흡광도치,비교각조대이와SGC적보호작용.결과 전염후BDNF-BMSC적Nestin、NSE、GFAP양성솔균고우미전염적BMSC(P치균<0.01).이와주사후재상동시간점각조간SGC밀도화흡광도치차이균구유통계학의의(P치균<0.05),기중BDNF-BMSC조SGC밀도최대、흡광도치최대;각조SGC밀도화흡광도치재술후4주시비술후1주화2주시하강(P치균<0.05).결론 돈서이와내이식BMSC、BDNF기인、BDNF-BMSC균가길항아미잡성대SGC적손상,기중BDNF-BMSC적보호작용최강.
Objective To investigate the protective role of brain-derived neurotrophic factor (BDNF) gene transfected bone-marrow mesenchymal stem cells (BMSC) on cochlear spiral ganglion cells (SGC) impaired by aminoglycoside antibiotics(AmAn). Methods The differentiation of BMSC transfected by BDNF gene (BDNF-BMSC) were detected with immunohistochemical examination of Nestin, neuronspecific enolase (NSE), and glial fibrillary acid protein (GFAP)antibody in vitro. BDNF gene transfected BMSC were transplanted into the cochleae of guinea pigs deafened by amikacin, while the control groups were designed in which artificial perilymphatic fluid(APF), BMSC or BDNF gene was injected into cochleae alone. The cochleae were obtained on the week 1,2 and 4 after injection, respectively, paraffin-embedded,and cut in a paramodiolar plane subsequently. The histopathological changes of cochleae were observed, the density of SGC was calculated by staining with HE, and the corresponding optical density (COD) was calculated with immunohistochemical staining using NSE antibody. And the protective role of various groups on the cochlear SGC were compared. Results The positive staining rate of BDNF gene transfected BMSC with Nestin, NSE and GFAP antibody were all higher than that of BMSC in vitro (P< 0.01). After transplantation into cochleae, the differences of SGC density and COD among various groups were all significant on the same time points(P < 0.05). The SGC density and COD of the BDNF gene transfected BMSC group were the highest. The SGC density and COD of various groups on week 4 were all obviously decreased than those on week 1 and 2 (P < 0.05). Conclusion AmAn-induced SGC damage could be depressed by BMSC, BDNF gene or BDNF gene transfected BMSC transplantion into cochleae, while BDNF gene transfected BMSC showed the best protective role.