風洞噪聲對小鼠聽性腦干反應和耳蝸超微結(jié)構(gòu)的影響及雌激素的防護作用
發(fā)布時間：2019-06-09 13:00
【摘要】：目的根據(jù)現(xiàn)場采集相關數(shù)據(jù)模擬風洞噪聲，建立小鼠噪聲性耳聾模型，觀察風洞噪聲對小鼠聽性腦干反應和耳蝸超微結(jié)構(gòu)的損傷。脫離風洞噪聲后給予雌激素干預，觀察雌激素對小鼠ABR閾值及耳蝸超微結(jié)構(gòu)的作用，探討雌激素對噪聲性聽力損傷的影響，為噪聲性耳聾的預防和治療提供依據(jù)。 方法40只昆明種小鼠（雄性）隨機分為單純噪聲組（A組）、雌激素治療組（B組），每組20只，分別暴露于模擬風洞噪聲環(huán)境中，每天8小時，連續(xù)7天。B組噪聲暴露結(jié)束后30min肌肉注射苯甲酸雌二醇0.15mg/只，而A組則不給予注射苯甲酸雌二醇。分別于實驗前、噪聲暴露3天、噪聲暴露7天、脫離噪聲后恢復3天、脫離噪聲后恢復7天共5個時間節(jié)點測試各組小鼠聽性腦干反應(ABR)閾值，并于噪聲暴露7天和脫離噪聲后恢復7天時將小鼠快速斷頭、迅速取出耳蝸，2.5％戊二醛固定液進行離體耳蝸灌注，并在固定液中固定4小時。(1)采用硬剝法暴露出基底膜，然后進行脫水、干燥、鍍金等處理，掃描電鏡觀察并拍照。(2)將標本浸入10％乙二胺四乙酸中脫鈣3周，然后進行脫水、包埋、固化、切片及染色處理，透射電鏡下觀察并拍照。 結(jié)果兩組小鼠實驗前ABR閾值無統(tǒng)計學差異，接觸噪聲后兩組小鼠均有明顯聽力損失，表現(xiàn)為閾值隨噪聲暴露時間延長而升高，噪聲暴露3天、噪聲暴露7天的ABR閾值均比實驗前增高，但兩組組間無統(tǒng)計學差異。當脫離噪聲后，兩組聽力均有不同程度恢復，脫離噪聲后恢復3天、脫離噪聲后恢復7天時雌激素治療組ABR閾值較單純噪聲組低，兩組間有統(tǒng)計學差異。 掃描電鏡顯示：在噪聲暴露7天后兩組小鼠耳蝸損傷嚴重，內(nèi)、外毛細胞限局性片狀廣泛缺失，Corti器塌陷、變形。脫離噪聲恢復7天時，單純噪聲組小鼠耳蝸內(nèi)、外毛細胞限局性點狀缺失，Corti器塌陷、皺縮、變形；而雌激素治療組小鼠耳蝸病變較輕，只有內(nèi)毛細胞輕度散亂、融合，外毛細胞基本正常。 透射電鏡顯示：在噪聲暴露7天后兩組小鼠耳蝸內(nèi)、外毛細胞胞漿內(nèi)均有較多空泡形成，細胞線粒體減少。脫離噪聲恢復7天時，單純噪聲組小鼠耳蝸外毛細胞線粒體大致正常，，基本無空泡，內(nèi)毛細胞雖有空泡形成，但較噪聲暴露7天時數(shù)量減少；雌激素治療組小鼠耳蝸內(nèi)毛細胞胞漿內(nèi)僅有很少量空泡形成，外毛細胞線粒體均正常，無空泡形成。 結(jié)論風洞噪聲能明顯損傷小鼠聽性腦干反應，其ABR閾值隨噪聲暴露時間延長而升高，脫離噪聲后兩組均有不同程度恢復，雌激素治療組恢復效果好于單純噪聲組。風洞噪聲同時造成小鼠耳蝸超微結(jié)構(gòu)的損傷，雌激素治療組損傷較單純噪聲組明顯減輕，與聽力檢查結(jié)果相吻合。雌激素對小鼠風洞噪聲性耳聾具有保護作用。
[Abstract]:Objective to establish a model of noise-induced deafness in mice according to the relevant data collected in the field, and to observe the damage of auditory brainstem response and cochlear ultrastructure induced by wind tunnel noise in mice. The effects of estrogen on ABR threshold and cochlear ultrastructure in mice were observed after breaking away from wind tunnel noise, and the effect of estrogen on noise-induced hearing loss was discussed, which provided the basis for the prevention and treatment of noise-induced deafness. Methods 40 Kunming mice (male) were randomly divided into simple noise group (group A) and estrogen treatment group (group B), with 20 mice in each group, exposed to simulated wind tunnel noise environment for 8 hours a day. For 7 days, group B received intramuscular injection of Estradiol benzoate 0.15mg/ into 30min after noise exposure, while Group A was not injected with Estradiol benzoate. Before the experiment, noise exposure for 3 days, noise exposure for 7 days, recovery from noise for 3 days, and recovery for 7 days after noise, five time nodes were used to test the (ABR) threshold of auditory brainstem response in each group. When the mice were exposed to noise for 7 days and recovered from noise for 7 days, the mice were quickly decapitated, the cochlea were quickly removed, and 2.5% glutaraldehyde fixation solution was perfused into the isolated cochlea. And fixed in the stationary solution for 4 hours. (1) the basement membrane was exposed by hard stripping method, then dehydrated, dried and gilded, observed and photographed by scanning electron microscope. (2) the specimens were immersed in 10% ethylenediamine tetraacetic acid for 3 weeks. Then dehydration, embedding, curing, slicing and staining were carried out, and observed and photographed under transmission electron microscope. Results there was no significant difference in ABR threshold between the two groups before the experiment. After exposure to noise, the mice in the two groups had obvious hearing loss, which showed that the threshold increased with the prolongation of noise exposure time and exposed to noise for 3 days. The ABR threshold of noise exposure for 7 days was higher than that before the experiment, but there was no significant difference between the two groups. When the noise was removed, the hearing of the two groups recovered to varying degrees, and the ABR threshold of the estrogen treatment group was lower than that of the simple noise group 3 days after the noise removal and 7 days after the noise removal, and there was significant difference between the two groups. Scanning electron microscope (SEM) showed that after 7 days of noise exposure, the cochlea of the two groups was seriously damaged, the localized flakes of inner and outer hair cells were widely deleted, and the Corti apparatus collapsed and deformed. After 7 days of recovery from noise, focal punctate deletion of outer hair cells, collapse, shrinkage and deformation of Corti apparatus were found in the cochlea of mice in the simple noise group. In the estrogen treatment group, the cochlear lesions were mild, only the inner hair cells were slightly scattered, fusion and the outer hair cells were basically normal. Transmission electron microscope showed that more vacuoles were formed in the cytoplasm of outer hair cells and the number of mitochondria decreased in the cochlea of the two groups after 7 days of noise exposure. When the isolated noise recovered for 7 days, the mitochondria of the outer hair cells of the cochlea in the noise group were about normal, and there were no vacuoles. Although the vacuoles were formed in the inner hair cells, the number of the inner hair cells was lower than that in the noise exposure group at 7 days. In estrogen treatment group, only a small number of vacuoles were formed in the cytoplasm of hair cells in cochlea of mice treated with estrogen, and the mitochondria of outer hair cells were normal and no vacuoles were formed. Conclusion Wind tunnel noise can significantly damage auditory brainstem response in mice, and the ABR threshold increases with the prolongation of noise exposure time. Both groups recovered to varying degrees after noise removal. The recovery effect of estrogen treatment group was better than that of simple noise group. Wind tunnel noise also caused the damage of cochlea ultrastructure in mice. The damage in estrogen treatment group was significantly less than that in simple noise group, which was consistent with the results of hearing examination. Estrogen has protective effect on wind tunnel noise induced deafness in mice.
【學位授予單位】：安徽醫(yī)科大學
【學位級別】：碩士
【學位授予年份】：2012
【分類號】：R764
本文編號：2495573
[Abstract]:Objective to establish a model of noise-induced deafness in mice according to the relevant data collected in the field, and to observe the damage of auditory brainstem response and cochlear ultrastructure induced by wind tunnel noise in mice. The effects of estrogen on ABR threshold and cochlear ultrastructure in mice were observed after breaking away from wind tunnel noise, and the effect of estrogen on noise-induced hearing loss was discussed, which provided the basis for the prevention and treatment of noise-induced deafness. Methods 40 Kunming mice (male) were randomly divided into simple noise group (group A) and estrogen treatment group (group B), with 20 mice in each group, exposed to simulated wind tunnel noise environment for 8 hours a day. For 7 days, group B received intramuscular injection of Estradiol benzoate 0.15mg/ into 30min after noise exposure, while Group A was not injected with Estradiol benzoate. Before the experiment, noise exposure for 3 days, noise exposure for 7 days, recovery from noise for 3 days, and recovery for 7 days after noise, five time nodes were used to test the (ABR) threshold of auditory brainstem response in each group. When the mice were exposed to noise for 7 days and recovered from noise for 7 days, the mice were quickly decapitated, the cochlea were quickly removed, and 2.5% glutaraldehyde fixation solution was perfused into the isolated cochlea. And fixed in the stationary solution for 4 hours. (1) the basement membrane was exposed by hard stripping method, then dehydrated, dried and gilded, observed and photographed by scanning electron microscope. (2) the specimens were immersed in 10% ethylenediamine tetraacetic acid for 3 weeks. Then dehydration, embedding, curing, slicing and staining were carried out, and observed and photographed under transmission electron microscope. Results there was no significant difference in ABR threshold between the two groups before the experiment. After exposure to noise, the mice in the two groups had obvious hearing loss, which showed that the threshold increased with the prolongation of noise exposure time and exposed to noise for 3 days. The ABR threshold of noise exposure for 7 days was higher than that before the experiment, but there was no significant difference between the two groups. When the noise was removed, the hearing of the two groups recovered to varying degrees, and the ABR threshold of the estrogen treatment group was lower than that of the simple noise group 3 days after the noise removal and 7 days after the noise removal, and there was significant difference between the two groups. Scanning electron microscope (SEM) showed that after 7 days of noise exposure, the cochlea of the two groups was seriously damaged, the localized flakes of inner and outer hair cells were widely deleted, and the Corti apparatus collapsed and deformed. After 7 days of recovery from noise, focal punctate deletion of outer hair cells, collapse, shrinkage and deformation of Corti apparatus were found in the cochlea of mice in the simple noise group. In the estrogen treatment group, the cochlear lesions were mild, only the inner hair cells were slightly scattered, fusion and the outer hair cells were basically normal. Transmission electron microscope showed that more vacuoles were formed in the cytoplasm of outer hair cells and the number of mitochondria decreased in the cochlea of the two groups after 7 days of noise exposure. When the isolated noise recovered for 7 days, the mitochondria of the outer hair cells of the cochlea in the noise group were about normal, and there were no vacuoles. Although the vacuoles were formed in the inner hair cells, the number of the inner hair cells was lower than that in the noise exposure group at 7 days. In estrogen treatment group, only a small number of vacuoles were formed in the cytoplasm of hair cells in cochlea of mice treated with estrogen, and the mitochondria of outer hair cells were normal and no vacuoles were formed. Conclusion Wind tunnel noise can significantly damage auditory brainstem response in mice, and the ABR threshold increases with the prolongation of noise exposure time. Both groups recovered to varying degrees after noise removal. The recovery effect of estrogen treatment group was better than that of simple noise group. Wind tunnel noise also caused the damage of cochlea ultrastructure in mice. The damage in estrogen treatment group was significantly less than that in simple noise group, which was consistent with the results of hearing examination. Estrogen has protective effect on wind tunnel noise induced deafness in mice.
【學位授予單位】：安徽醫(yī)科大學
【學位級別】：碩士
【學位授予年份】：2012
【分類號】：R764
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