【摘要】：目的檢測(cè)順鉑損傷后，神經(jīng)分化因子NeuroD在大鼠耳蝸螺旋神經(jīng)節(jié)的表達(dá)變化，探討其在螺旋神經(jīng)節(jié)損傷后的修復(fù)作用研究。 方法成年SD雄性大鼠64只，按隨機(jī)數(shù)隨機(jī)分為4組，分別為：對(duì)照組：按體重以0.9％生理鹽水5ml/kg腹腔注射，連續(xù)5天，1次/天，在第6天處死；用藥1天組：按體重以順鉑（山東齊魯）5mg/kg腹腔注射，在第2天處死；用藥3天組：按體重以順鉑（山東齊魯）5mg/kg腹腔注射，連續(xù)3天，1次/天，在第4天處死；用藥5天組：按體重以順鉑（山東齊魯）5mg/kg腹腔注射，連續(xù)5天，1次/天，，在第6天處死。每組16只，建立順鉑耳毒性模型。通過(guò)HE染色、免疫熒光染色、免疫組織化學(xué)染色、免疫印跡（WesternBlot）及實(shí)時(shí)定量PCR（Real-Time PCR）等手段，檢測(cè)用順鉑造聾的耳蝸螺旋神經(jīng)節(jié)損傷后不同的時(shí)間點(diǎn)NeuroD的表達(dá)變化。 結(jié)果成功建立順鉑耳毒性大鼠模型，隨著用藥時(shí)間的延長(zhǎng)，神經(jīng)分化因子NeuroD在耳蝸螺旋神經(jīng)節(jié)中呈動(dòng)態(tài)變化。NeuroD的mRNA表達(dá)在用藥1天及3天組，與對(duì)照組及用藥5d組比較差異具有統(tǒng)計(jì)學(xué)意義（P值均0.01）；免疫組化圖片光密度檢測(cè)結(jié)果顯示用藥1天組及3天組與對(duì)照組比較具有統(tǒng)計(jì)學(xué)意義(P0.01)，用藥5天組與對(duì)照組比較無(wú)統(tǒng)計(jì)學(xué)意義（P0.05）；Western Blot結(jié)果顯示用藥1天組及3天組與對(duì)照組比較具有統(tǒng)計(jì)學(xué)意義(P0.01)，用藥5天組與對(duì)照組比較無(wú)統(tǒng)計(jì)學(xué)意義（P0.05）。 結(jié)論NeuroD在用藥1天后開(kāi)始增加，3天后達(dá)到高峰，5天后下降；在用藥早期有一過(guò)性表達(dá)增強(qiáng)，后期表達(dá)下降同時(shí)聽(tīng)力損失明顯。表明NeuroD可能參與順鉑損傷螺旋神經(jīng)節(jié)后的修復(fù)過(guò)程。
[Abstract]:Objective to investigate the expression of differentiation factor NeuroD in cochlear spiral ganglion (SCG) after cisplatin injury. Methods Sixty-four adult SD male rats were randomly divided into four groups: control group: the control group was injected intraperitoneally with 0.9% normal saline 5ml/kg for 5 days, once a day, and was killed on the 6th day. The rats in the treatment group were given intraperitoneal injection of cisplatin (Shandong Qilu) 5mg/kg by intraperitoneal injection on the second day, and those in the 3-day group received intraperitoneal injection of cisplatin (Shandong Qilu) 5mg/kg according to body weight for 3 days, once a day, and executed on the 4th day. In the 5-day group, cisplatin (Qilu, Shandong) 5mg/kg was injected intraperitoneally for 5 days, once a day, and executed on the 6th day. The ototoxicity model of cisplatin was established with 16 rats in each group. The expression of NeuroD at different time points after cochlear helical ganglion injury induced by cisplatin was detected by means of HE staining, immunofluorescence staining, immunohistochemical staining, immunoblotting (WesternBlot) and real-time quantitative PCR (Real-Time PCR). Results the rat model of cisplatin ototoxicity was successfully established. With the prolongation of administration time, the neural differentiation factor NeuroD was dynamically changed in the cochlear spiral ganglion. The mRNA expression of NeuroD was observed on the 1st and 3rd day of administration. There was significant difference between the two groups (P < 0.01). The results of light density detection showed that the light density of the treatment group and the 3 day group were significantly higher than that of the control group (P0.01), but there was no significant difference between the 5-day group and the control group (P0.05). The results of Western Blot showed that there was significant difference between the 1 day group and 3 day group compared with the control group (P0.01), but there was no significant difference between the 5 day group and the control group (P0.05). Conclusion NeuroD began to increase after 1 day treatment, reached its peak after 3 days, and decreased after 5 days. The results suggest that NeuroD may be involved in the repair of spiral ganglion injured by cisplatin.
【學(xué)位授予單位】：第四軍醫(yī)大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2012
【分類(lèi)號(hào)】：R764

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