發(fā)布時間：2018-08-16 15:28

【摘要】：目的:建立犬聲帶瘢痕模型,分析損傷后不同時間段內(nèi)聲帶愈合的形態(tài)學(xué)、組織病理學(xué)及細(xì)胞外基質(zhì)(ECM)相關(guān)影響因子的變化。方法:5只實驗犬隨機分為對照組(1只)和實驗組(4只),對照組不做特殊處理,實驗組行支撐喉鏡下CO_2激光雙側(cè)聲帶燒灼術(shù)。各只實驗犬分別在5個不同時間(術(shù)前和術(shù)后6h、3周、8周和12周)于支撐喉鏡下觀察聲帶損傷的大體形態(tài)學(xué)變化后處死,取材。采用蘇木精-伊紅染色、免疫熒光等方法觀察聲帶各層組織病理學(xué)變化和超微形態(tài)學(xué)變化。觀測聲帶組織內(nèi)賴氨酰氧化酶(LOX)、熱休克蛋白70(HSP70)和透明質(zhì)酸(HA)的累積光密度值(IOD)的變化規(guī)律。結(jié)果:1喉內(nèi)鏡示,損傷后3周聲帶創(chuàng)面輕度充血水腫及炎性滲出;術(shù)后8周創(chuàng)面充血水腫消失,局部攣縮凹陷,無粘連和肉芽形成;術(shù)后12周雙側(cè)聲帶表面光滑,局部攣縮,聲帶瘢痕形成,聲嘶明顯。2蘇木精-伊紅染色示,術(shù)后6h聲帶創(chuàng)面大量炎性細(xì)胞浸潤、紅細(xì)胞滲出,細(xì)胞充血水腫;術(shù)后3周聲帶創(chuàng)面成纖維細(xì)胞增生,新生血管形成,大量纖維組織無序排列;術(shù)后8周聲帶創(chuàng)面各層纖維組織增生紊亂,血管增粗;術(shù)后12周時,聲帶創(chuàng)面固有層增厚,大量膠原纖維增多,排列紊亂,呈團狀或束狀,纖維間較多不規(guī)則間隙;血管和腺體少見或消失。3免疫熒光示LOX、HSP70、HA均定位于細(xì)胞質(zhì)和細(xì)胞核,術(shù)后不同時間段表達的陽性程度不同,大多在炎性細(xì)胞、血管內(nèi)皮細(xì)胞和腺體內(nèi)表達較強烈。4IOD值比較,在不同時間段LOX的IOD值之間均有差異(P0.05);HSP70在術(shù)前與術(shù)后12周的IOD值無明顯異常,余各組間差異有統(tǒng)計學(xué)意義(P0.01);HA在術(shù)后3周與12周間無異常,余各組間差異有統(tǒng)計學(xué)意義(P0.05)。5LOX在術(shù)后6h表達減弱,術(shù)后3~12周呈上升趨勢;HSP70在術(shù)后6h升至峰值,術(shù)后3~12周呈下降趨勢;HA在術(shù)后6h表達減弱,術(shù)后8周升至峰值,術(shù)后8~12周下降。6透射電鏡顯示,聲帶術(shù)后3~8周固有層成纖維細(xì)胞極度活躍狀態(tài),細(xì)胞腫脹,細(xì)胞器豐富;術(shù)后12周時,聲帶固有層內(nèi)仍功能活躍,纖維細(xì)胞呈網(wǎng)狀排列紊亂,纖維間隙較大。結(jié)論:支撐喉鏡下CO_2激光聲帶燒灼術(shù)建立犬聲帶瘢痕模型可靠,損傷后3個月聲帶瘢痕基本形成。LOX、HSP70、HA在聲帶瘢痕形成的早中晚期發(fā)揮著不同的作用,可以作為檢測聲帶瘢痕形成較敏感的指標(biāo)。
[Abstract]:Aim: to establish canine vocal cord scar model and analyze the changes of morphology histopathology and related factors of extracellular matrix (ECM) related to vocal cord healing in different time after injury. Methods five experimental dogs were randomly divided into two groups: control group (1 dog) and experimental group (4 dogs). The control group did not receive special treatment. The experimental group received CO_2 laser cauterization of bilateral vocal cords under laryngoscope. The experimental dogs were killed at 5 different times (3 weeks before and 6 hours after operation, 8 weeks and 12 weeks after operation) after observing the morphological changes of vocal cord injury under supporting laryngoscope. The histopathological and ultrastructural changes of vocal cord were observed by hematoxylin-eosin staining and immunofluorescence. The changes of cumulative optical density (IOD) of (LOX), heat shock protein 70 (HSP70) and hyaluronic acid (HA) in vocal cord tissues were observed. Results 1 laryngeal endoscopy showed slight hyperemia and edema and inflammatory exudation of vocal cord wound at 3 weeks after injury, hyperemia and edema disappeared at 8 weeks after injury, no adhesion and granulation formation in local contracture, smooth surface of bilateral vocal cord and local contracture at 12 weeks after operation. The vocal cord scar formation, hoarseness obvious .2 hematoxylin-eosin staining showed that the vocal cord wound surface was infiltrated by a large number of inflammatory cells, red blood cell exudation, hyperemia and edema at 6 hours after operation, fibroblast proliferation and angiogenesis of vocal cord wound at 3 weeks after operation. At 8 weeks after operation, the lamina propria of vocal cord wound was thickened, and a large number of collagenous fibers were arranged in disorder, in the shape of mass or bundles, the lamina propria of vocal cord wound was thickened and a large number of collagenous fibers were increased at 12 weeks after operation. There were many irregular interspaces between the fibers. 3. 3 immunofluorescence showed that Loxus HSP70HA was located in cytoplasm and nucleus, and the positive degree was different at different time after operation, most of which were in inflammatory cells. The expression of IOD in endothelial cells and glands was significantly higher than that in LOX at different time points (P0.05). The IOD values of HSP70 were not abnormal before and 12 weeks after operation. There was no significant difference in HA between 3 and 12 weeks after operation (P0.01). There was a significant difference between the other groups (P0.05) .5LOX expression decreased at 6 hours after operation, and the HSP70 increased to a peak at 6 hours after operation at 3 and 12 weeks after operation. The expression of HA decreased at 6 hours after operation and rose to the peak at 8 weeks after operation. A decrease of .6 transmission electron microscope showed that fibroblasts in lamina propria were extremely active, cells were swollen and organelle was abundant at 3 and 8 weeks after vocal cord operation. At 12 weeks after operation, the vocal cord lamina propria was still active, the fibrous cells were disorganized and the fiber gap was larger. Conclusion: the canine vocal cord scar model established by CO_2 laser cauterization under bracing laryngoscope is reliable. The cicatricial formation of vocal cord in 3 months after injury. Loxo HSP70HA plays different roles in the early, middle and late stage of vocal cord scar formation. It can be used as a sensitive index for the detection of vocal cord scar formation.
【作者單位】： 武漢大學(xué)附屬同仁醫(yī)院武漢市第三醫(yī)院耳鼻咽喉科;
【基金】：武漢市衛(wèi)計委中青年醫(yī)學(xué)骨干培養(yǎng)計劃工程[No:武衛(wèi)生計生(2014)77號文件]
【分類號】：R-332;R767.4

【相似文獻】

相關(guān)期刊論文 前7條

1 王燕;鄧仁躍;;人聲帶瘢痕的組織學(xué)特點[J];聽力學(xué)及言語疾病雜志;2010年04期

2 吳修法;徐潔潔;;應(yīng)用肝細(xì)胞生長因子凝膠修復(fù)慢性聲帶瘢痕[J];聽力學(xué)及言語疾病雜志;2011年04期

3 甄瑞清;魏春生;;透明質(zhì)酸對聲帶瘢痕的預(yù)防作用[J];中國眼耳鼻喉科雜志;2013年06期

4 王家佳;魏春生;;聲帶瘢痕的形成及治療進展[J];聽力學(xué)及言語疾病雜志;2011年03期

5 許偉民;顧志明;劉運章;;纖維喉鏡聲帶手術(shù)后并發(fā)聲帶瘢痕的預(yù)防及其可行性探討[J];臨床耳鼻咽喉頭頸外科雜志;2007年06期

6 潘葉挺;鄒堅定;龔梁;陳建強;;喉外傷后聲帶瘢痕形成機制的實驗研究[J];中國現(xiàn)代醫(yī)生;2012年29期

7 ;[J];;年期

相關(guān)會議論文 前4條

1 傅德慧;黃永望;;聲帶瘢痕患者發(fā)聲及嗓音調(diào)節(jié)能力空氣動力學(xué)研究[A];全國耳鼻咽喉頭頸外科中青年學(xué)術(shù)會議論文匯編[C];2012年

2 傅德慧;黃永望;;聲帶瘢痕患者發(fā)聲及嗓音調(diào)節(jié)能力空氣動力學(xué)研究[A];2014年首次中國中西醫(yī)結(jié)合耳鼻咽喉科中青年學(xué)術(shù)會議論文匯編[C];2014年

3 傅德慧;;聲帶瘢痕患者發(fā)聲及嗓音調(diào)節(jié)能力空氣動力學(xué)研究[A];2014年首次中國中西醫(yī)結(jié)合耳鼻咽喉科中青年學(xué)術(shù)會議論文匯編[C];2014年

4 張雷;趙麗娜;姜秀文;;大鼠聲帶瘢痕動物模型的建立及價值[A];2006年浙江省耳鼻咽喉科學(xué)術(shù)會議論文匯編[C];2006年

，

本文編號：2186394