中华航海医学与高气压医学杂志
中華航海醫學與高氣壓醫學雜誌
중화항해의학여고기압의학잡지
CHINESE JOURNAL OF NAUTICAL MEDICINE AND HYPERBARIC MEDICINE
2014年
3期
163-167
,共5页
戴尉君%夏国明%张健%吴建%范静平
戴尉君%夏國明%張健%吳建%範靜平
대위군%하국명%장건%오건%범정평
Math1基因%神经生长因子β基因%噪声%螺旋神经节%基因治疗
Math1基因%神經生長因子β基因%譟聲%螺鏇神經節%基因治療
Math1기인%신경생장인자β기인%조성%라선신경절%기인치료
Math 1 gene%Nerve growth factor gene%Noise%Spiral ganglion cells%Gene therapy
目的 观察Math1(mouse atonal homologue 1 gene,Math1)基因及神经生长因子β基因(nerve growth factor beta gene,NGFβ)共转染对噪声性听力损伤豚鼠耳蜗螺旋神经节细胞的保护作用.方法 选用40只健康成年白色纯种豚鼠制备稳态噪声(110 dB SPL)耳聋模型,任选爆震前后听阈阈移大于60 dB SPL的动物作为实验动物,共选35只,采用数字表法随机分为4组,其中Ⅰ组10只为双基因组,导入腺病毒介导的双基因Ad-Math 1/NGFβ;Ⅱ组10只为Math1组,导入腺病毒介导的单基因Ad-Math1;Ⅲ组10只为NGF组,导入腺病毒介导的单基因Ad-NGFβ;Ⅳ组5只为对照组,导入腺病毒;另选健康成年白色纯种豚鼠5只为正常组,不接受噪声和基因转染.基因转染后第1周,取豚鼠耳蜗组织固定切片行免疫组织化学染色;基因转染后第4周,取豚鼠耳蜗组织固定切片行HE染色.结果 基因转染后1周,Ad-Math 1/NGFβ在豚鼠耳蜗内成功转染,耳蜗各回膜性组织中均有表达,且表达强度基本相等;基因转染后第4周HE染色显示:Ⅰ组豚鼠耳蜗组织中螺旋神经节细胞数平均为(166.4 ±2.58)个,明显多于Ⅱ组(92.1±1.34)个、Ⅲ组(88.6±1.22)个,差异均有统计学意义(P＜0.01),且细胞形态与正常细胞形态相近.结论 Math1联合NGFβ基因共转染在噪声损伤后豚鼠耳蜗中高效表达,对噪声性听力损伤豚鼠耳蜗螺旋神经节细胞有明显的保护作用.
目的 觀察Math1(mouse atonal homologue 1 gene,Math1)基因及神經生長因子β基因(nerve growth factor beta gene,NGFβ)共轉染對譟聲性聽力損傷豚鼠耳蝸螺鏇神經節細胞的保護作用.方法 選用40隻健康成年白色純種豚鼠製備穩態譟聲(110 dB SPL)耳聾模型,任選爆震前後聽閾閾移大于60 dB SPL的動物作為實驗動物,共選35隻,採用數字錶法隨機分為4組,其中Ⅰ組10隻為雙基因組,導入腺病毒介導的雙基因Ad-Math 1/NGFβ;Ⅱ組10隻為Math1組,導入腺病毒介導的單基因Ad-Math1;Ⅲ組10隻為NGF組,導入腺病毒介導的單基因Ad-NGFβ;Ⅳ組5隻為對照組,導入腺病毒;另選健康成年白色純種豚鼠5隻為正常組,不接受譟聲和基因轉染.基因轉染後第1週,取豚鼠耳蝸組織固定切片行免疫組織化學染色;基因轉染後第4週,取豚鼠耳蝸組織固定切片行HE染色.結果 基因轉染後1週,Ad-Math 1/NGFβ在豚鼠耳蝸內成功轉染,耳蝸各迴膜性組織中均有錶達,且錶達彊度基本相等;基因轉染後第4週HE染色顯示:Ⅰ組豚鼠耳蝸組織中螺鏇神經節細胞數平均為(166.4 ±2.58)箇,明顯多于Ⅱ組(92.1±1.34)箇、Ⅲ組(88.6±1.22)箇,差異均有統計學意義(P＜0.01),且細胞形態與正常細胞形態相近.結論 Math1聯閤NGFβ基因共轉染在譟聲損傷後豚鼠耳蝸中高效錶達,對譟聲性聽力損傷豚鼠耳蝸螺鏇神經節細胞有明顯的保護作用.
목적 관찰Math1(mouse atonal homologue 1 gene,Math1)기인급신경생장인자β기인(nerve growth factor beta gene,NGFβ)공전염대조성성은력손상돈서이와라선신경절세포적보호작용.방법 선용40지건강성년백색순충돈서제비은태조성(110 dB SPL)이롱모형,임선폭진전후은역역이대우60 dB SPL적동물작위실험동물,공선35지,채용수자표법수궤분위4조,기중Ⅰ조10지위쌍기인조,도입선병독개도적쌍기인Ad-Math 1/NGFβ;Ⅱ조10지위Math1조,도입선병독개도적단기인Ad-Math1;Ⅲ조10지위NGF조,도입선병독개도적단기인Ad-NGFβ;Ⅳ조5지위대조조,도입선병독;령선건강성년백색순충돈서5지위정상조,불접수조성화기인전염.기인전염후제1주,취돈서이와조직고정절편행면역조직화학염색;기인전염후제4주,취돈서이와조직고정절편행HE염색.결과 기인전염후1주,Ad-Math 1/NGFβ재돈서이와내성공전염,이와각회막성조직중균유표체,차표체강도기본상등;기인전염후제4주HE염색현시:Ⅰ조돈서이와조직중라선신경절세포수평균위(166.4 ±2.58)개,명현다우Ⅱ조(92.1±1.34)개、Ⅲ조(88.6±1.22)개,차이균유통계학의의(P＜0.01),차세포형태여정상세포형태상근.결론 Math1연합NGFβ기인공전염재조성손상후돈서이와중고효표체,대조성성은력손상돈서이와라선신경절세포유명현적보호작용.
Objective To observe the protective effect of Math1/NGFβ (Mouse atonal homologue Ⅰ gene/ nerve growth factor beta gene) gene transfection on spiral ganglion cells in guinea pigs with noise-induced hearing damage.Methods Forty heahhy white guinea pigs were used to develop the model of stable noiseinduced hearing damage (110 dB SPL).Thirty-five animals with hearing threshold greater than 60 dB SPL both before and after burst explosion were chosen as experimental animals,and then were randomly divided into 4 groups:groups Ⅰ,Ⅱ,Ⅲ and Ⅳ.The animals in group Ⅰ (n =10) (or the Mathl/NGFβ group) were inoculated with Ad-Math1/NGFβ,the animals in group Ⅱ (n =10) or (the Math1 group) were imported with Ad-Mathl,the animals in group Ⅲ(n =10) (or the NGF group) were imported with Ad-NGFβ,the animals in group Ⅳ (or the control group) were imported with adenovirus,and still another 5 healthy white guinea pigs were chosen as the normal group,which had neither noise exposure nor gene transfection.Cochlea sections of guinea pigs were collected for immunohistochemical staining,respectively at week 1 after gene transfection.Then,at week 4 after gene transfection,cochlea sections of guinea pigs were collected for HE staining.Results At week 1 after gene transfection,genes of Ad-Math1/NGFβ were successfully transfected in the cochlea of guinea pigs and expressions of Ad-Math1/NGFβ protein could be detected in each turn of the cochlea,with expression intensity being essentially the same.HE staining at week 4 after gene transfection indicated that the number of spiral ganglions in the Math1/NGFβ group was on the average (166.4 ± 2.58),which was significantly greater than those of group Ⅱ (92.1 ± 1.34) and group Ⅲ (88.6 ± 1.22)),with statistical significance (P ＜ 0.01).Conclusions Ad-Math1/NGFβ could be expressed at a high level in the noiseimpaired cochlea,suggesting that Ad-Math1/NGFβ seemed to have a protective effect on cochlear spiral ganglion cells following noise-impaired hearing damage.
