本文選題：血管外膜 + 血凝酶��； 參考：《安徽醫(yī)科大學(xué)》2014年碩士論文

【摘要】：目的 圍手術(shù)期血管滲血及術(shù)后創(chuàng)口滲血是外科手術(shù)過程中常見的并發(fā)癥，因而減少術(shù)中及術(shù)后失血在臨床上越來越受到重視。目前臨床上常用的止血方法有藥物止血法和理化止血法等，，但目前關(guān)于這兩種止血方法對(duì)組織修復(fù)長(zhǎng)期觀察的研究較少。本實(shí)驗(yàn)旨在通過對(duì)新西蘭大白兔頸總動(dòng)脈外膜剝離的方法造成血管滲血，繼而用血凝酶以及電灼兩種方法止血，以觀察血凝酶（hemocoagulase atrox, HA）與電灼(electric cauterization, EC)局部止血對(duì)動(dòng)脈修復(fù)的影響，用以指導(dǎo)臨床應(yīng)用。 方法 健康純種新西蘭大白兔27只，均約20周齡，雌雄各半，體重2.5~3.0kg，按觀察時(shí)間隨機(jī)分為3組：0d組（n=9）、14d組（n=9）、28d組（n=9），采用頸動(dòng)脈外膜剝離的方法造成血管滲血模型，每組再根據(jù)止血方法不同隨機(jī)分成3個(gè)亞組：對(duì)照組（control，CON組，n=3）、血凝酶組（HA組，n=3）和電灼組（EC組，n=3）。CON組家兔兩側(cè)頸總動(dòng)脈剝離外膜后用生理鹽水（normal saline, NS）紗布止血，HA組和EC組在外膜剝離后分別予以HA（濃度為2U/ml）沾濕的紗布及電灼（功率為40w）止血。0d組在術(shù)后直接取出兩側(cè)頸總動(dòng)脈，14d組和28d組分別術(shù)后正常飲食相應(yīng)天數(shù)后麻醉家兔，活體、無菌條件下取出兩側(cè)動(dòng)脈。將取出的血管標(biāo)本分為三段：①約0.5cm標(biāo)本制成蠟塊，用HE染色和MASSON三色膠原染色觀察不同亞組動(dòng)脈修復(fù)及重塑的病理學(xué)改變，免疫組化染色檢測(cè)動(dòng)脈單核細(xì)胞趨化蛋白-1（MCP-1）、轉(zhuǎn)化生長(zhǎng)因子-β1（TGF-β1）及血管內(nèi)皮生長(zhǎng)因子（VEGF）蛋白的表達(dá)情況；②約1cm標(biāo)本用透射電鏡觀察不同組別動(dòng)脈血管細(xì)胞分化情況；③約1cm標(biāo)本用逆轉(zhuǎn)錄-聚合酶鏈反應(yīng)（RT-PCR）法檢測(cè)不同組別標(biāo)本中MCP-1、TGF-β1及VEGF的mRNA表達(dá)情況。 結(jié)果 1、所有家兔均成功造成滲血模型，各組動(dòng)脈滲血程度基本相當(dāng)，各組家兔一般狀況及手術(shù)情況差異均無統(tǒng)計(jì)學(xué)意義。 2、NS、HA及EC止血對(duì)動(dòng)脈外膜剝離后血管重塑的影響 病理學(xué)觀察顯示0d CON組及HA組HE及Masson染色見外膜已被去除，內(nèi)膜中膜形態(tài)完好，而EC組見中膜彈力纖維及膠原纖維結(jié)構(gòu)疏松，偶見內(nèi)皮細(xì)胞且形態(tài)不規(guī)則。術(shù)后14d可見三組血管外膜增生活躍，內(nèi)膜增厚，管腔面積縮小，膠原纖維密度明顯增大。HA組動(dòng)脈中、內(nèi)膜厚度、膠原纖維相對(duì)密度（collagen fibers density，CFD）及狹窄率(stenotic ratio, SR)均小于EC組（P0.05），管腔面積及血管重塑指數(shù)(remodeling index, RI)均大于EC組（P0.05），而HA組同CON組相比無統(tǒng)計(jì)學(xué)意義（P0.05）；28d HA組各指標(biāo)均顯著優(yōu)于EC組（P0.05），而與CON組無明顯差別。 3、生理鹽水、血凝酶及電灼止血對(duì)動(dòng)脈細(xì)胞增值、分化的影響 透射電鏡觀察顯示0d CON組及HA組見內(nèi)皮細(xì)胞完整，平滑肌細(xì)胞形態(tài)正常；而EC組見內(nèi)皮細(xì)胞空泡樣變性，平滑肌細(xì)胞水腫嚴(yán)重；術(shù)后14d各組新生外膜可見大量微絲及粗面內(nèi)質(zhì)網(wǎng)擴(kuò)張的成纖維細(xì)胞（Fibroblasts, F），呈現(xiàn)出肌成纖維細(xì)胞（Myofibroblasts, MF）特征，并有向內(nèi)膜遷徙的趨勢(shì)；28dCON組及HA組外膜F已逆轉(zhuǎn)為MF，而EC組仍可見F。 4、生理鹽水、血凝酶及電灼止血對(duì)動(dòng)脈MCP-1、TGF-β1及VEGF表達(dá)的影響 免疫組化染色及RT-PCR結(jié)果顯示：0、14及28d CON組和HA組MCP-1、TGF-β1及VEGF陽性細(xì)胞及mRNA表達(dá)無明顯差異，且28d兩組表達(dá)已下降近0d水平（P0.05）；而EC組同CON組相比，術(shù)后14及28d MCP-1、TGF-β1及VEGF蛋白及mRNA表達(dá)差異均有統(tǒng)計(jì)學(xué)意義（P0.05）。 結(jié)論 在動(dòng)脈外膜剝離滲血模型中，血凝酶止血可有效減輕損傷血管的炎癥反應(yīng)，降低血管重塑程度和狹窄情況，與單純的鹽水紗布止血血管修復(fù)結(jié)果一致，是一種安全、有效、更接近自然狀況的局部止血藥物。
[Abstract]:objective
Perioperative vascular osmotic blood and postoperative wound oozing are common complications in the surgical procedure, thus reducing blood loss in the operation and post operation is becoming more and more important. Currently, the commonly used hemostatic methods are drug hemostasis and physicochemical hemostasis, but the two methods of hemostasis have been observed for a long time. The aim of this study was to investigate the effects of hemostasis on the arterial repair by two methods: hemocoagulase atrox (HA) and electrocautery (electric cauterization, EC).
Method
27 healthy and pure New Zealand white rabbits were divided into 3 groups of 20 weeks old and half female and male and body weight 2.5~3.0kg. According to the observation time, they were randomly divided into 3 groups: group 0d (n=9), group 14d (n=9) and group 28d (n=9). The blood vessel infiltration model was caused by exfoliation of carotid artery, and each group was randomly divided into 3 subgroups according to the different hemostasis methods: the control group (control, CON group, n=3), The hemagglutinin group (group HA, n=3) and electrocautery group (group EC, n=3).CON group of rabbits were treated with normal saline (normal saline, NS) gauze after exfoliation of the lateral common carotid artery in group.CON. The HA group and EC group were treated with HA (concentration for 2U/ml) wet gauze and electric cauterization after exfoliation of the outer membrane. The two arteries were taken out of the rabbits, the living body and the aseptic condition after the normal diet of the 28d group. The blood vessel specimens were divided into three segments: 1. About 0.5cm specimens were made of wax blocks. The pathological changes of the repair and remodeling of different subgroups were observed by HE staining and MASSON trichromatic collagen staining, and the arterial monomer was detected by immunohistochemical staining. Nuclear cell chemoattractant protein -1 (MCP-1), transforming growth factor - beta 1 (TGF- beta 1) and vascular endothelial growth factor (VEGF) protein expression. (2) about 1cm specimens were observed by transmission electron microscopy to observe the differentiation of vascular cells in different groups of arteries; and about 1cm specimens were detected by reverse transcription polymerase chain reaction (RT-PCR) method for the detection of MCP-1, TGF- beta 1 in different groups. And the mRNA expression of VEGF.
Result
1, all rabbits were successful in oozing the blood model. The degree of arterial bleeding in each group was basically the same.
2, the effects of NS, HA and EC hemostasis on vascular remodeling after epicardial detachment.
Pathological observation showed that 0d CON group and HA group HE and Masson staining showed that the outer membrane had been removed, the membrane morphology of the intima was intact, and the middle membrane elastic fibers and collagen fibers were loose in the EC group, and the endothelial cells were irregular in morphology. After the operation, the three groups of vascular adventitial hyperplasia were active, the inner membrane was thickened, the area of the lumen was reduced, and the collagen fiber density was obvious In the.HA group, the thickness of the intima, the relative density of collagen fiber (collagen fibers density, CFD) and the stenosis rate (stenotic ratio, SR) were all smaller than that of the EC group (P0.05), and the lumen area and the vascular remodeling index (remodeling index) were all larger than those in the group. It was better than group EC (P0.05), but no significant difference from group CON.
3, the effects of physiological saline, hemocoagulase and electrocautery on the proliferation and differentiation of arterial cells.
The transmission electron microscope showed that the endothelial cells were intact in 0d CON group and HA group, and the morphology of smooth muscle cells was normal, while in group EC, the endothelial cells were vacuolated and the smooth muscle cells were edema. After the operation, a large number of microfilaments and dilated fibroblasts of rough endoplasmic reticulum (F) were found in all 14d groups, and myofibroblast (Myofi) was presented. Broblasts, MF) had the characteristics of migration to the intima, while the outer membrane F of group 28dCON and group HA had been reversed to MF, while F. was still visible in EC group.
4, the effects of normal saline, hemagglutinase and electrocautery on the expression of MCP-1, TGF- beta 1 and VEGF in arteries
Immunohistochemical staining and RT-PCR results showed that there was no significant difference between 0,14 and 28d CON group and HA group MCP-1, TGF- beta 1 and VEGF positive cells and mRNA expression, and the expression of 28d two decreased near 0d level (P0.05).
conclusion
In the exfoliation model of epicardial exfoliation, hemostasis can effectively reduce the inflammatory reaction of damaged vessels, reduce the degree of vascular remodeling and stenosis, and is consistent with the results of simple saline gauze repair blood vessel repair. It is a safe, effective, and closer to the natural state of the local hemostatic drugs.
【學(xué)位授予單位】：安徽醫(yī)科大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2014
【分類號(hào)】：R619.1

【參考文獻(xiàn)】

相關(guān)期刊論文 前7條

1 湯月霞;劉永;王華;梁春;陳金明;吳宗貴;;血管外膜剝離致內(nèi)膜增生病變的動(dòng)物模型[J];中國(guó)動(dòng)脈硬化雜志;2008年01期

2 注射用血凝酶安全性回顧研究協(xié)作組;單愛蓮;;外科手術(shù)中應(yīng)用注射用血凝酶安全性多中心大樣本回顧性調(diào)查[J];中國(guó)新藥與臨床雜志;2011年12期

3 傅宏義;周磊;;蛇毒類凝血酶的研究進(jìn)展[J];中國(guó)藥學(xué)雜志;2008年04期

4 王宏波;張軍;魯愛俠;丁光輝;黃術(shù)勝;;三種不同止血方法對(duì)腹部切口愈合的影響[J];中國(guó)現(xiàn)代藥物應(yīng)用;2010年18期

5 郭明珂;張英澤;田德虎;吳嘯波;陳占法;馬利杰;鄭占樂;;局部應(yīng)用5-氟尿嘧啶抑制移植靜脈內(nèi)膜增生的實(shí)驗(yàn)研究[J];中國(guó)修復(fù)重建外科雜志;2009年08期

6 ;Experimental and clinical study of influence of high-frequency electric surgical knives on healing of abdominal incision[J];World Journal of Gastroenterology;2006年25期

7 韋軍民;朱明煒;張忠濤;賈振庚;何小東;萬遠(yuǎn)廉;唐大年;葉國(guó)棟;孫瑞元;;尖吻蝮蛇血凝酶對(duì)腹部手術(shù)切口止血作用的有效性和安全性[J];中國(guó)新藥雜志;2007年14期

本文編號(hào)：1907205