發(fā)布時(shí)間：2018-04-20 09:17

  本文選題：水楊酸鈉 + caspase-3�。� 參考：《廣西醫(yī)科大學(xué)》2010年碩士論文

【摘要】： 目的研究水楊酸鈉(sodium salicylate, SS)誘導(dǎo)豚鼠耳蝸螺旋神經(jīng)節(jié)細(xì)胞(spiral ganglion neuron, SGN)凋亡及凋亡的機(jī)制。 方法40只成年花色雄性黑目豚鼠隨機(jī)分為4組,每組10只。A組:空白對(duì)照組;B組:人工外淋巴液(artificial perilymph, APL)組(左耳經(jīng)圓窗龕注入APL后未給SS);C組:SS(400 mg·kg~(-1)·d~(-1))+APL組(左耳經(jīng)圓窗龕注入APL后經(jīng)腹腔SS給藥);D組:SS(400 mg·kg~(-1)·d~(-1))+zDEVD-FMK組(左耳經(jīng)圓窗龕注入zDEVD-FMK后經(jīng)腹腔SS給藥)。所有動(dòng)物均于給藥前后分別進(jìn)行聽性腦干反應(yīng)(auditory brainstem response, ABR)閾值測(cè)試。10 % SS(溶于APL配制而成)均采用腹腔注射給藥方式,400 mg·kg~(-1)·d~(-1),并于連續(xù)給藥10天后即時(shí)處死動(dòng)物取出耳蝸,采用脫氧核糖核苷酸末端轉(zhuǎn)移酶介導(dǎo)的缺口末端標(biāo)記法(terminal-deoxynucleotidyl transferase mediated dUTP nick end labeling, TUNEL)檢測(cè)耳蝸SGN凋亡細(xì)胞,免疫組織化學(xué)染色方法檢測(cè)SGN內(nèi)激活型caspase-3蛋白表達(dá)情況,透射電鏡(transmission electron microscopy, TEM)觀察SGN超微結(jié)構(gòu)變化。 結(jié)果ABR測(cè)試結(jié)果顯示A、B兩組動(dòng)物聽閾均無(wú)明顯變化,C組動(dòng)物給藥后聽閾提高,與A、B兩組比較有顯著性差異(p0.01,p0.01),D組動(dòng)物聽閾較C組降低(p0.05);TUNEL標(biāo)記顯示A、B兩組動(dòng)物耳蝸中均未見SGN凋亡細(xì)胞,亦無(wú)激活型caspase-3蛋白表達(dá),而C組則出現(xiàn)較多TUNEL陽(yáng)性著色SGN,并伴有激活型caspase-3表達(dá)的增高,D組較C組TUNEL陽(yáng)性著色SGN減少,激活型caspase-3表達(dá)亦降低,A、B兩組與C、D兩組比較差異均有顯著統(tǒng)計(jì)學(xué)意義(p0.01,p0.01),而C、D兩組比較亦有顯著差別(p0.01);透射電鏡觀察結(jié)果顯示A、B兩組SGN超微結(jié)構(gòu)正常,C組SGN伴有明顯細(xì)胞凋亡形態(tài)學(xué)特征,D組SGN超微結(jié)構(gòu)未見明顯凋亡征象。 結(jié)論①SS可誘導(dǎo)豚鼠耳蝸SGN發(fā)生凋亡;②SGN凋亡與caspase-3激活有關(guān);③zDEVD-FMK可在一定程度上拮抗caspase-3的激活,并阻斷其引起的SGN凋亡。
[Abstract]:Objective to investigate the mechanism of sodium salicylate (SS) induced apoptosis of spiral ganglion cells (SGNN) in guinea pig cochlea. Methods Forty adult male black guinea pigs were randomly divided into 4 groups. 10 rats in each group: control group B: artificial perilymphs (left ear after injection of APL through a round window niche without APL) APL group (left ear was injected with APL through a circular window niche and intraperitoneal administration of SS: 400 mg / kg) zDEVD-FMK group (left ear group: 400 mg / kg ~ (-1)) zDEVD-FMK group (left ear: n = 400 mg / kg 路kg ~ (-1)) / d ~ (-1) / d ~ (-1) / d ~ (-1) / d ~ (-1) / d ~ (-1) / d ~ (-1) / d ~ (-1) / d ~ (-1) / d ~ (-1) / d ~ (-1) / d ~ (-1) / d ~ (-1) / d ~ (-1). ZDEVD-FMK was injected through a round window niche and then intraperitoneally administered with SS. All animals were given auditory brainstem response (ABRs) threshold test. 10% SS (dissolved in APL) was administered intraperitoneally with 400 mg / kg ~ (-1) DX respectively. Cochlea were removed immediately after 10 days of continuous administration, and cochlea were taken out immediately after 10 days of continuous administration. Terminal deoxynucleotidyl transferase mediated dUTP nick end labeling (Tunel) was used to detect SGN apoptotic cells in cochlea, and the expression of activated caspase-3 protein in SGN was detected by immunohistochemical staining. The ultrastructural changes of SGN were observed by transmission electron microscopy (TM). Results the results of ABR test showed that there was no significant change in hearing threshold in both groups. There was a significant difference between group A and group A (P 0.01). Compared with group C, the auditory threshold of group D was significantly lower than that of group C. No apoptotic SGN cells were found in cochlea of group A and B, and no expression of activated caspase-3 protein was found in the cochlea of the two groups. In group C, there were more TUNEL positive staining SGNs, and increased expression of activated caspase-3 in group D was lower than that in group C in TUNEL positive staining SGN. The expression of activated caspase-3 was also decreased in both groups. There was a significant difference in the expression of activated caspase-3 between group A and C, and the difference between group C and group C was significant (p 0.01), and there was also a significant difference between group C and group C (P 0.01). Transmission electron microscopy showed that the ultrastructure of SGN in group A B was normal and that in group C was significantly higher than that in group C (P < 0.05). The ultrastructure of SGN in group D showed no obvious signs of apoptosis. Conclusion 1SS can induce apoptosis of SGN in guinea pig cochlea. The apoptosis of 2SGN is related to the activation of caspase-3. 1SS can antagonize the activation of caspase-3 to some extent and block the apoptosis of SGN induced by 1SS.
【學(xué)位授予單位】：廣西醫(yī)科大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2010
【分類號(hào)】：R764

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本文編號(hào)：1777175