本文關(guān)鍵詞： 增生性瘢痕 內(nèi)皮抑素 兔耳瘢痕模型 靶向治療　出處：《浙江大學(xué)》2013年博士論文　論文類型：學(xué)位論文

【摘要】：研究背景： 增生性瘢痕的發(fā)生發(fā)展與血管形成密切相關(guān),抑制傷口愈合后的過度新生血管有助于減少增生性瘢痕的形成。因此,尋找一種能夠靶向作用于瘢痕組織抑制瘢痕形成的藥物有一定的意義。文獻報道內(nèi)皮抑素局部注射能抑制瘢痕增生,但是全身用藥能否抑制瘢痕增生尚未見文獻報道；內(nèi)皮抑素抑制瘢痕增生的作用機制目前仍不清楚。 目的： 研究內(nèi)皮抑素全身用藥對兔耳瘢痕的作用并初步探討其可能的作用機制。 研究方法： 選取體重在2.2-2.5kg的新西蘭大白兔共8只,隨機分為實驗組和對照組,每組4只,分籠飼養(yǎng)兩周。靜脈麻醉后用角膜鉆在8只新西蘭白兔的耳朵腹側(cè)建立直徑7mm的創(chuàng)面(每耳6個,共96個)。棉球壓迫止血,待傷口自行愈合,一周后麻醉下用鑷子剝?nèi)ヰ杵?造成新的傷口,自行愈合。在兔耳鉆孔術(shù)后兩周開始,實驗組每天腹腔注射內(nèi)皮抑素,對照組每天腹腔注射等量的生理鹽水,連續(xù)用藥20天。在藥物注射當(dāng)天和注射后第6,13,20天(即傷后15,21,28,35天)觀察兔耳瘢痕顏色、質(zhì)地變化,并用游標(biāo)卡尺測量瘢痕厚度；在以上各個時點分別用激光多普勒血流儀對實驗組與對照組瘢痕組織微循環(huán)血流灌注進行檢測,研究分析內(nèi)皮抑素對兔耳瘢痕微循環(huán)血流灌注的影響。在給藥后20天(即傷后第35天),將實驗動物麻醉,分別切取實驗組與對照組瘢痕組織。從瘢痕組織頂部正中將瘢痕用利刀切為兩半,一半放入10%甲醛溶液中保存留作HE染色及]masson染色,另一半放入液氮瓶中并保存在-80℃的深低溫冰箱內(nèi)留作分子生物學(xué)檢測Ⅰ型膠原和凋亡抑制基因bcl-2蛋白。 結(jié)果： 1大體檢測 1.1瘢痕顏色 對照組的瘢痕充血明顯,呈紫色或暗紅色,而實驗組瘢痕充血不明顯,呈淺紅色或偏蒼白。 1.2瘢痕厚度 傷后15天兩組瘢痕厚度達(dá)到高峰,兩組厚度之間在統(tǒng)計學(xué)上沒有顯著性差異。之后,隨著時間的推移,兩組瘢痕厚度均逐漸下降。但是,實驗組厚度下降更快,在傷后第21天及35天兩組的瘢痕厚度在統(tǒng)計學(xué)上具有顯著差異(P0.05)。 1.3激光多普勒測定瘢痕血流灌注 對照組兔耳瘢痕內(nèi)的血流灌注指標(biāo)PU值呈先上升后逐漸下降,實驗組的PU值則呈逐漸下降走勢,在傷后21天兩者的PU值相比在統(tǒng)計學(xué)上有顯著性差異(P0.05)。 2組織學(xué)檢測 2.1瘢痕增生指數(shù)(SEI) 實驗組的瘢痕增生指數(shù)(1.094±0.19)低于對照組(1.364±0.28),兩組SEI相比在統(tǒng)計學(xué)上具有顯著性差異(P0.01) 2.2瘢痕微血管密度(MVD) 通過HE染色下微血管計數(shù),連續(xù)計數(shù)5個400倍視野中的微血管數(shù),取平均值,計算微血管密度。實驗組的微血管密度為1.734±0.94個/高倍視野,對照組為5.634±1.78個/高倍視野,實驗組MVD低于對照組,在統(tǒng)計學(xué)上具有顯著性差異(P0.01) 2.3瘢痕Masson染色 實驗組瘢痕組織內(nèi)膠原較細(xì)、排列整齊,對照組膠原粗大且排列紊亂。 3分子生物學(xué)檢測 3.1Ⅰ型膠原檢測 實驗組瘢痕組織中Ⅰ型膠原的表達(dá)低于對照組。 3.2凋亡抑制基因bcl-2蛋白檢測 實驗組的bcl-2表達(dá)低于對照組。 結(jié)論： 內(nèi)皮抑素全身用藥可以抑制兔耳增生性瘢痕的形成,表現(xiàn)為同一時點較對照組瘢痕厚度減小、瘢痕充血減輕、微血管密度減少、膠原排列較規(guī)則,其機制可能是內(nèi)皮抑素通過下調(diào)bcl-2促進瘢痕組織內(nèi)皮細(xì)胞凋亡進而抑制瘢痕新生血管形成,進一步使內(nèi)皮細(xì)胞釋放的促進纖維化的細(xì)胞因子減少從而間接抑制瘢痕增生；然而,內(nèi)皮抑素對成纖維細(xì)胞作用的具體作用機制尚待進一步實驗研究。
[Abstract]:Research background:
Closely related with the occurrence and development of vascular hypertrophic scar formation, inhibit excessive neovascularization after wound healing can help reduce scar formation. Therefore, to find a way to have a certain significance of targeting drugs to inhibit scar formation of scar tissue. Reported local injection of endostatin can inhibit scar hyperplasia, but systemic medication can inhibit scar hyperplasia has not been reported; mechanism of endostatin inhibit scar hypertrophy remains unclear.
Objective:
The effect of endostatin on the rabbit ear scar was studied and its possible mechanism was preliminarily discussed.
Research methods:
Select the weight in 2.2-2.5kg New Zealand white rabbits were 8, were randomly divided into experimental group and control group, 4 rats in each group, divided into cages for two weeks. After intravenous anesthesia with corneal wound to establish drill diameter 7mm in 8 New Zealand white rabbits ear ventral (6 per ear, a total of 96). SpongeBob oppression check the blood, until the wound healed after a week under anesthesia with tweezers stripped crust, resulting in a new wound, healing. At the beginning of the two week after the rabbit ear drilling operation, the experimental group received intraperitoneal injection of endostatin, the control group normal saline was injected every day, 20 days of continuous use. In the day of drug injection and the first 6,13,20 days after injection (15,21,28,35 days after injury) observation of rabbit ear scar color, texture changes, and use vernier caliper to measure the thickness of the scar; above each time point respectively by laser Doppler flowmetry in experimental group and control group. The scar tissue perfusion Measurement, analysis of effects of endostatin on rabbit ear scar microcirculation perfusion. After administration for 20 days (i.e., thirty-fifth days after injury), experimental animal anesthesia, were harvested for the experimental group and control group. The scar tissue from the center of the top of the scar scar tissue with a sharp knife cut in half, half in the left HE staining and]masson staining to save 10% Formaldehyde Solution, the other half bottle and saved in liquid nitrogen for type I collagen and apoptosis detection of molecular biology of suppressor gene Bcl-2 protein in deep low temperature refrigerator -80 degrees.
Result錛，

本文編號：1531608