肝素在動(dòng)物實(shí)驗(yàn)外傷性鼓膜穿孔愈合中的作用
發(fā)布時(shí)間：2018-12-12 04:08
【摘要】： 研究背景：外傷性鼓膜穿孔是臨床常見(jiàn)的疾病,常因直接或間接外力作用所致,雖大部分可自行愈合,但一旦感染,將成為永久性穿孔。鼓膜貼補(bǔ)法簡(jiǎn)單,安全,廉價(jià),近年來(lái)貼補(bǔ)藥物的選擇一直是研究的熱點(diǎn)。表皮生長(zhǎng)因子(EGF)作為貼補(bǔ)藥物治療鼓膜穿孔的作用已被證實(shí)。在動(dòng)物實(shí)驗(yàn)中,國(guó)內(nèi)已有學(xué)者通過(guò)肉眼觀察應(yīng)用肝素作為貼補(bǔ)藥物,可促進(jìn)外傷性鼓膜穿孔的愈合。鼓膜穿孔愈合過(guò)程需要多種體內(nèi)細(xì)胞因子的參與,如表皮生長(zhǎng)因子(EGF)、轉(zhuǎn)化生長(zhǎng)因子-α(TGF-α)、堿性成纖維細(xì)胞生長(zhǎng)因子(bFGF)、血小板衍生生長(zhǎng)因子(PDGF)、角質(zhì)形成細(xì)胞生長(zhǎng)因子(KGF)等。研究發(fā)現(xiàn)在創(chuàng)傷愈合過(guò)程中上述生長(zhǎng)因子都以與肝素結(jié)合的形式存在,不僅可提高生長(zhǎng)因子生物學(xué)效應(yīng),而且可以保護(hù)其免于被各種理化因素滅活。肝素有利于創(chuàng)傷修復(fù)已被國(guó)內(nèi)外學(xué)者所證實(shí)。肝素在鼓膜穿孔中對(duì)轉(zhuǎn)化生長(zhǎng)因子-α(TGF-α)、堿性成纖維細(xì)胞生長(zhǎng)因子(bFGF)的影響至今尚未見(jiàn)報(bào)道。探討在外傷性鼓膜穿孔愈合過(guò)程中肝素對(duì)生長(zhǎng)因子TGF-α、bFGF的影響進(jìn)而促進(jìn)鼓膜穿孔的愈合是本研究的重點(diǎn)。 研究目的：探討局部應(yīng)用肝素在外傷性鼓膜穿孔愈合中的作用。 研究方法：①建立大鼠外傷性鼓膜穿孔模型：Wistar大鼠50只,所有動(dòng)物先用戊巴比妥鈉腹腔麻醉(0.3ml／100g),在電耳鏡下用灼熱針尖在大鼠雙側(cè)鼓膜后下象限制成約2mm×2mm緊張部的圓形穿孔。術(shù)后用氨芐青霉素(150mg／kg)肌肉注射預(yù)防感染,每天一次,連續(xù)5天。②實(shí)驗(yàn)方法：50只大鼠隨機(jī)分為兩組,第一組25只左耳外耳道置無(wú)菌干棉球；右耳肝素明膠海綿貼補(bǔ)。第二組25只左耳EGF明膠海綿貼補(bǔ)；右耳肝素明膠海綿貼補(bǔ)。分別于術(shù)后1天、3天、5天、7天和10天時(shí),每組處死5只大鼠,解剖顳骨,剝離聽(tīng)泡外側(cè)壁,取出完整鼓膜。對(duì)取得的鼓膜標(biāo)本用10%甲醛固定,石蠟包埋,切片,常規(guī)HE染色,采用免疫組化兩步法檢測(cè)。免疫組化結(jié)果用配對(duì)樣本(同體比較)的t檢驗(yàn),以P0.05為差異具有統(tǒng)計(jì)學(xué)意義。 研究結(jié)果： 1.光鏡下肝素組三天時(shí)上皮細(xì)胞增厚,細(xì)胞由扁平細(xì)胞變成多邊形,細(xì)胞間距增加,核質(zhì)比增大,上皮層與纖維層分離,上皮增生明顯。纖維層也增厚,達(dá)5-6層,內(nèi)層柱狀上皮也增生。五天時(shí)鼓膜已基本愈合,且結(jié)構(gòu)與正常鼓膜接近。 2.免疫組化圖像分析,肝素組與空白組相比TGF-α、bFGF表達(dá)值差異有統(tǒng)計(jì)學(xué)意義,p0.05。肝素組與EGF組之間TGF-α、bFGF表達(dá)值差異無(wú)統(tǒng)計(jì)學(xué)意義,P0.05。 研究結(jié)論： 1.肝素可提高TGF-α、bFGF在鼓膜穿孔愈合中的表達(dá),并延長(zhǎng)其表達(dá)時(shí)間。 2.局部應(yīng)用肝素可縮短穿孔鼓膜的愈合時(shí)間,促進(jìn)穿孔愈合。 3.肝素貼補(bǔ)法治療鼓膜穿孔,愈合更接近正常鼓膜組織結(jié)構(gòu)。
[Abstract]:Background: traumatic tympanic membrane perforation is a common clinical disease, often caused by direct or indirect external force. Tympanic membrane patch is simple, safe and cheap. The effect of epidermal growth factor (EGF) on tympanic membrane perforation has been confirmed. In animal experiments, heparin is used as a patching drug to promote the healing of traumatic tympanic membrane perforation. The healing process of tympanic membrane perforation requires a variety of cytokines, such as epidermal growth factor (EGF), transforming growth factor- 偽 (TGF- 偽), basic fibroblast growth factor (bFGF), platelet-derived growth factor (PDGF), Keratinocyte growth factor (KGF). It was found that these growth factors existed in the form of binding with heparin during wound healing, which not only enhanced the biological effects of growth factors, but also protected them from being inactivated by various physical and chemical factors. Heparin is beneficial to wound repair and has been confirmed by domestic and foreign scholars. The effect of heparin on transforming growth factor- 偽 (TGF- 偽) and basic fibroblast growth factor (bFGF) during tympanic membrane perforation has not been reported. To investigate the effect of heparin on growth factor TGF- 偽, bFGF during the healing of traumatic tympanic membrane perforation, and to promote the healing of tympanic membrane perforation is the focus of this study. Objective: to investigate the effect of local application of heparin on the healing of traumatic tympanic membrane perforation. Methods: 1 the rat model of traumatic tympanic membrane perforation was established: 50 Wistar rats were treated with pentobarbital sodium intraperitoneal anesthesia (0.3ml/100g). A circular perforation of the 2mm 脳 2mm tension region was limited to the posterior inferior image of the bilateral tympanic membrane with a scorching needle under the ear mirror. After operation, ampicillin (150mg/kg) was injected intramuscularly to prevent infection, once a day for 5 days. Methods: 50 rats were randomly divided into two groups: the first group (25 rats) had sterile dry cotton balls in the external auditory canal of the left ear, and the right ear heparin gelatin sponge patch. Group 2: EGF gelatin sponge patch in left ear and heparin gelatin sponge patch in right ear. At 1 day, 3 days, 5 days, 7 days and 10 days after operation, 5 rats in each group were killed, temporal bone was dissected, lateral wall of auditory vesicle was removed and intact tympanic membrane was removed. The tympanic membrane samples were fixed with 10% formaldehyde, embedded in paraffin, sectioned and stained with conventional HE, and detected by immunohistochemical two-step method. Immunohistochemical results of paired samples (compared with the t test), with P0.05 as the difference was statistically significant. Results: 1. In the heparin group, the epithelial cells became thicker, the cells changed from flat cells to polygons, the cell spacing increased, the ratio of nucleus to cytoplasm increased, the epithelial layer separated from the fibrous layer, and the epithelium proliferated obviously. The fibrous layer also thickened to 5-6 layers and the inner columnar epithelium also proliferated. At 5 days, the tympanic membrane was basically healed and its structure was close to that of the normal tympanic membrane. 2. Immunohistochemical analysis showed that the expression of TGF- 偽 and bFGF in heparin group was significantly different from that in blank group (p 0.05). There was no significant difference in the expression of TGF- 偽 and bFGF between heparin group and EGF group (P 0.05). Conclusion: 1. Heparin increased the expression of TGF- 偽 and bFGF in the healing of tympanic membrane perforation and prolonged the expression time. 2. Local application of heparin can shorten the healing time of perforated tympanic membrane and promote perforation healing. 3. Heparin patch therapy for tympanic membrane perforation, healing closer to the normal tympanic tissue structure.
【學(xué)位授予單位】：山西醫(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
本文編號(hào)：2373873
[Abstract]:Background: traumatic tympanic membrane perforation is a common clinical disease, often caused by direct or indirect external force. Tympanic membrane patch is simple, safe and cheap. The effect of epidermal growth factor (EGF) on tympanic membrane perforation has been confirmed. In animal experiments, heparin is used as a patching drug to promote the healing of traumatic tympanic membrane perforation. The healing process of tympanic membrane perforation requires a variety of cytokines, such as epidermal growth factor (EGF), transforming growth factor- 偽 (TGF- 偽), basic fibroblast growth factor (bFGF), platelet-derived growth factor (PDGF), Keratinocyte growth factor (KGF). It was found that these growth factors existed in the form of binding with heparin during wound healing, which not only enhanced the biological effects of growth factors, but also protected them from being inactivated by various physical and chemical factors. Heparin is beneficial to wound repair and has been confirmed by domestic and foreign scholars. The effect of heparin on transforming growth factor- 偽 (TGF- 偽) and basic fibroblast growth factor (bFGF) during tympanic membrane perforation has not been reported. To investigate the effect of heparin on growth factor TGF- 偽, bFGF during the healing of traumatic tympanic membrane perforation, and to promote the healing of tympanic membrane perforation is the focus of this study. Objective: to investigate the effect of local application of heparin on the healing of traumatic tympanic membrane perforation. Methods: 1 the rat model of traumatic tympanic membrane perforation was established: 50 Wistar rats were treated with pentobarbital sodium intraperitoneal anesthesia (0.3ml/100g). A circular perforation of the 2mm 脳 2mm tension region was limited to the posterior inferior image of the bilateral tympanic membrane with a scorching needle under the ear mirror. After operation, ampicillin (150mg/kg) was injected intramuscularly to prevent infection, once a day for 5 days. Methods: 50 rats were randomly divided into two groups: the first group (25 rats) had sterile dry cotton balls in the external auditory canal of the left ear, and the right ear heparin gelatin sponge patch. Group 2: EGF gelatin sponge patch in left ear and heparin gelatin sponge patch in right ear. At 1 day, 3 days, 5 days, 7 days and 10 days after operation, 5 rats in each group were killed, temporal bone was dissected, lateral wall of auditory vesicle was removed and intact tympanic membrane was removed. The tympanic membrane samples were fixed with 10% formaldehyde, embedded in paraffin, sectioned and stained with conventional HE, and detected by immunohistochemical two-step method. Immunohistochemical results of paired samples (compared with the t test), with P0.05 as the difference was statistically significant. Results: 1. In the heparin group, the epithelial cells became thicker, the cells changed from flat cells to polygons, the cell spacing increased, the ratio of nucleus to cytoplasm increased, the epithelial layer separated from the fibrous layer, and the epithelium proliferated obviously. The fibrous layer also thickened to 5-6 layers and the inner columnar epithelium also proliferated. At 5 days, the tympanic membrane was basically healed and its structure was close to that of the normal tympanic membrane. 2. Immunohistochemical analysis showed that the expression of TGF- 偽 and bFGF in heparin group was significantly different from that in blank group (p 0.05). There was no significant difference in the expression of TGF- 偽 and bFGF between heparin group and EGF group (P 0.05). Conclusion: 1. Heparin increased the expression of TGF- 偽 and bFGF in the healing of tympanic membrane perforation and prolonged the expression time. 2. Local application of heparin can shorten the healing time of perforated tympanic membrane and promote perforation healing. 3. Heparin patch therapy for tympanic membrane perforation, healing closer to the normal tympanic tissue structure.
【學(xué)位授予單位】：山西醫(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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