血流動(dòng)力學(xué)變化對(duì)血管重構(gòu)調(diào)控機(jī)制初步實(shí)驗(yàn)研究
發(fā)布時(shí)間：2019-06-04 07:06
【摘要】：研究背景許多臨床現(xiàn)象顯示,機(jī)械應(yīng)激是影響機(jī)體修復(fù)的重要因素,減輕機(jī)械應(yīng)激可以減輕修復(fù)的程度。在臨床工作中發(fā)現(xiàn),瘢痕的形成和機(jī)械應(yīng)激密切相關(guān),無機(jī)械應(yīng)激的手術(shù)切口瘢痕增生不明顯,骨折端的輕微機(jī)械應(yīng)激導(dǎo)致骨痂大量增生,即過度的修復(fù),無機(jī)械應(yīng)激可以獲得一期愈合,沒有骨痂形成等等,上述事實(shí)都說明機(jī)械應(yīng)激是影響組織修復(fù)的重要因素。并且很多相關(guān)的基礎(chǔ)研究也證實(shí)此理論,此理論能否在血管上得到證實(shí),血流動(dòng)力學(xué)的改變是否能夠?qū)е卵鼙诮Y(jié)構(gòu)的重構(gòu)?本課題組的前期工作中發(fā)現(xiàn)：1、噴涂a-氰基丙烯酸酯粘合劑均勻包被血管能夠達(dá)到固定血管吻合口的目的。2、噴涂粘合劑血管吻合口的狹窄率較低。眾多文獻(xiàn)報(bào)道靜脈血管移植到動(dòng)脈后,靜脈血管管壁出現(xiàn)了動(dòng)脈化的改變。除營養(yǎng)改變外,血流動(dòng)力學(xué)改變應(yīng)該是導(dǎo)致管壁發(fā)生變化的主要因素。另外當(dāng)前相關(guān)研究大多集中于臨床較為常見的靜脈移到動(dòng)脈后血流動(dòng)力學(xué)變化引起血管狹窄機(jī)制的研究,但對(duì)于其它血流動(dòng)力學(xué)變化調(diào)控血管重構(gòu)機(jī)制研究較少,本研究以動(dòng)靜脈移植和動(dòng)靜脈瘺兩種模型為對(duì)象,對(duì)血流動(dòng)力學(xué)變化調(diào)控血管重構(gòu)機(jī)制進(jìn)行初步實(shí)驗(yàn)研究。目的為了進(jìn)一步完善血流動(dòng)力學(xué)變化調(diào)控血管重構(gòu)機(jī)制理論,并對(duì)骨骼肌肉系統(tǒng)出現(xiàn)的“機(jī)械應(yīng)激促進(jìn)增生,減輕機(jī)械應(yīng)激抑制增生”現(xiàn)象,是否也適用于血流動(dòng)力學(xué)變化調(diào)控血管重構(gòu)進(jìn)行驗(yàn)證。我們選擇了當(dāng)前研究較少的兩種模型進(jìn)行相關(guān)理論初步實(shí)驗(yàn)研究,以動(dòng)靜脈移植和動(dòng)靜脈瘺兩種模型為對(duì)象,對(duì)血流動(dòng)力學(xué)參數(shù)進(jìn)行測(cè)量,對(duì)血管重構(gòu)中大體、組織、細(xì)胞及分子生物水平進(jìn)行觀察研究,并對(duì)兩要素之間關(guān)系進(jìn)行分析。方法(1)實(shí)驗(yàn)動(dòng)物及血管部位的選擇,比格犬和SD大鼠各5只,分別測(cè)量兩側(cè)頸總動(dòng)脈和頸靜脈以及股動(dòng)靜脈的外徑和便于操作有效長度,分析比較適合模型建立的實(shí)驗(yàn)動(dòng)物和血管。(2)動(dòng)靜脈移植模型建立,血流動(dòng)力學(xué)變化調(diào)控血管重構(gòu)機(jī)制研究。實(shí)驗(yàn)動(dòng)物為比格犬12只,12-15kg。1將犬左側(cè)頸總動(dòng)脈1cm切斷移植到右側(cè)頸外靜脈,結(jié)扎左側(cè)頸總動(dòng)脈,右側(cè)頸總動(dòng)脈為對(duì)照。分別于術(shù)后1,2,4周對(duì)兩組血管進(jìn)行大體觀察;HE染色,病理組織學(xué)觀測(cè)內(nèi)膜增生厚度；EVG染色,組織病理學(xué)觀測(cè)彈性纖維和膠原纖維含量變化；免疫組化染色觀測(cè)增值細(xì)胞陽性表達(dá);TUNEL技術(shù)觀測(cè)調(diào)亡細(xì)胞陽性表達(dá);RT-PCR技術(shù)觀測(cè)MMP2,9和eNOS基因表達(dá)。(3)動(dòng)靜脈瘺模型建立,血流動(dòng)力學(xué)變化調(diào)控血管重構(gòu)機(jī)制研究。實(shí)驗(yàn)動(dòng)物為比格犬12只,12-15kg。1將犬左側(cè)頸總動(dòng)脈和頸外靜脈傳統(tǒng)針線吻合,右側(cè)頸總動(dòng)脈和頸外靜脈為對(duì)照。分別于術(shù)后1,2,4周對(duì)兩組血管進(jìn)行大體觀察；HE染色,病理組織學(xué)觀測(cè)內(nèi)膜增生厚度；EVG染色,組織病理學(xué)觀測(cè)彈性纖維和膠原纖維含量變化;免疫組化染色觀測(cè)增值細(xì)胞陽性表達(dá);TUNEL技術(shù)觀測(cè)調(diào)亡細(xì)胞表達(dá);RT-PCR技術(shù)觀測(cè)MMP2,9和eNOS基因表達(dá)。結(jié)果(1)實(shí)驗(yàn)動(dòng)物比格犬較SD大鼠頸部血管更有利于手術(shù)操作,比格犬更易測(cè)量血流動(dòng)力學(xué)變化情況,比格犬血壓和人更加接近。(2)動(dòng)脈移植到靜脈后,血流速度和血壓明顯減小;HE染色觀測(cè)到內(nèi)膜厚度減小;EVG染色觀測(cè)到膠原纖維含量明顯減少,彈性纖維含量變化無統(tǒng)計(jì)學(xué)意義；免疫組化染色,觀測(cè)到PCNA指數(shù)無明顯變化;凋亡染色,觀測(cè)到凋亡細(xì)胞明顯增加,凋亡指數(shù)增大;電鏡觀察,中膜平滑肌細(xì)胞減少;RT-PCR觀測(cè)到MMP2,9和eNOS均出現(xiàn)基因表達(dá)增加。(3)動(dòng)靜脈瘺術(shù)后,靜脈和動(dòng)脈段血流速度增加,靜脈血壓上升,動(dòng)脈血壓小幅度降低;HE染色,測(cè)量動(dòng)脈段內(nèi)膜厚度增加,靜脈段內(nèi)膜厚度增加明顯;EVG染色觀測(cè)到動(dòng)脈段膠原纖維含量增加,靜脈段膠原纖維含量明顯增加,彈性纖維含量變化無統(tǒng)計(jì)學(xué)意義；免疫組化染色行觀測(cè)計(jì)算,動(dòng)脈段PCNA指數(shù)增大,靜脈段PCNA指數(shù)明顯增大;凋亡染色后觀測(cè)到動(dòng)脈段和靜脈段凋亡細(xì)胞無明顯增加,凋亡指數(shù)無明顯增大；電鏡觀察,動(dòng)脈段內(nèi)膜和中膜平滑肌細(xì)胞增多,靜脈段內(nèi)膜和中膜平滑肌細(xì)胞明顯增加;RT-PCR觀測(cè)到動(dòng)脈段MMP2,9和eON9表達(dá)減少,靜脈段MMP2,9和eONS表達(dá)明顯減少。結(jié)論通過建立動(dòng)靜脈移植模型和動(dòng)靜脈痿模型,從血流動(dòng)力學(xué)變化調(diào)控血管重構(gòu)機(jī)制角度進(jìn)行實(shí)驗(yàn)研究,進(jìn)一步完善相關(guān)理論。血流動(dòng)力學(xué)改變后,機(jī)械應(yīng)力信號(hào)傳導(dǎo)到血管壁細(xì)胞,酶(MMP2,9和eONS等)及細(xì)胞因子的表達(dá)出現(xiàn)變化,血管壁細(xì)胞增殖凋亡及遷移機(jī)制改變,細(xì)胞外基質(zhì)(膠原纖維和彈性纖維等)代謝機(jī)制改變,表現(xiàn)出血管壁結(jié)構(gòu)的變化。進(jìn)一步證明血流動(dòng)力學(xué)變化對(duì)血管重構(gòu)的調(diào)控作用,并且血流動(dòng)力學(xué)改變程度不同,血管重構(gòu)出現(xiàn)明顯不同變化。同時(shí)“機(jī)械應(yīng)激促進(jìn)增生,減輕機(jī)械應(yīng)激抑制增生”理論假設(shè)在血管重構(gòu)過程中進(jìn)一步得到驗(yàn)證。
[Abstract]:The research background has shown that the mechanical stress is an important factor that affects the repair of the body, and the degree of repair can be reduced by reducing the mechanical stress. During the clinical work, the formation of the mechanical stress and the mechanical stress are closely related, the non-mechanical stress-free operation incision is not obvious, the slight mechanical stress at the end of the fracture leads to a large amount of hyperplasia of the bone, that is, the excessive repair, the non-mechanical stress can obtain the first-stage healing, No bone formation and so on, the fact that mechanical stress is an important factor that affects tissue repair. And many related basic studies have also confirmed this theory, can this theory be confirmed on the blood vessel, whether the change of hemodynamics can lead to the reconstruction of the vessel wall structure? It was found in the earlier work of the research group that:1. The uniform coating of a-cyanoacrylate adhesive on the blood vessel can achieve the purpose of fixing the blood vessel anastomosis. A large number of literature reported that the vascular wall of the vein had an arterial change after the vein graft was transplanted to the artery. In addition to that change in nutrition, the change in hemodynamics should be a major factor in the change in the tube wall. in addition, most of that current study focus on the study of the mechanism of vascular stenosis due to the change in the hemodynamic change of the clinically more common vein to the post-arterial blood flow, but less research on the regulation of the vascular remodeling mechanism for other hemodynamic changes, In this study, two models of the arteriovenous graft and the arteriovenous graft were used as the target, and the mechanism of the blood flow dynamic change and the regulation of the vascular remodeling was studied. Objective In order to further improve the theory of blood flow dynamics and control the mechanism of vascular remodeling, and to study the "Mechanical stress promotes proliferation and relieves mechanical stress and inhibits proliferation" of the skeletal muscle system, it is also applicable to the changes of blood flow dynamics to control the vascular remodeling. In this paper, we selected two models of the current study to study the correlation theory. The two models of the arteriovenous graft and the arteriovenous graft were used as the target, the parameters of the blood flow were measured, and the general, the tissue, the cell and the molecular biological level in the blood vessel reconstruction were observed and studied. And the relationship between the two elements is analyzed. Methods (1) The external diameter of the common carotid artery and the jugular vein and the femoral vein and the effective length of the femoral vein were measured respectively in the experimental animals and the blood vessel, and the experimental animals and the blood vessels established by the model were compared. (2) The establishment of the arteriovenous graft model, the regulation of the blood flow dynamics and the mechanism of the reconstruction of the blood vessel. The experimental animals were 12-15kg beagle dogs.1 cm of the left common carotid artery of the dog was cut and transplanted to the right external jugular vein, and the left common carotid artery and the right common carotid artery were ligated. The two groups of blood vessels were generally observed at 1,2 and 4 weeks after operation, and the thickness of the intimal hyperplasia was observed by HE staining and pathology. The changes of the content of the elastic fibers and the collagen fibers were observed by the histopathological observation, and the positive expression of the value-added cells was observed by immunohistochemical staining. The expression of MMP2,9 and eNOS gene was observed by RT-PCR. (3) The establishment of the arteriovenous blood vessel model, the regulation of the blood flow dynamics and the mechanism of the reconstruction of the blood vessel. The experimental animals were 12-15kg beagle dogs. The left common carotid artery and the external jugular vein of the dog were matched with the traditional needle-and-needle, and the right-hand common carotid artery and the external jugular vein were used as the control. The two groups of blood vessels were generally observed at 1,2 and 4 weeks after operation, and the thickness of the intimal hyperplasia was observed by HE staining and pathology. The changes of the content of the elastic fibers and the collagen fibers were observed by the histopathological observation, and the positive expression of the value-added cells was observed by immunohistochemical staining. The expression of MMP2,9 and eNOS gene was observed by RT-PCR. Results (1) The experimental animal beagle dog was more beneficial to the operation of the neck vessel in the SD rat, and the beagle dogs were more likely to measure the hemodynamic changes, and the beagle dogs were closer to the blood pressure and human blood pressure. (2) The blood flow velocity and blood pressure were significantly reduced after the artery was transplanted to the vein; the thickness of the intima was observed with HE staining; the content of the collagen fibers was significantly reduced by the EVG staining, and the content of the elastic fibers was not statistically significant; and the expression of PCNA was observed without significant change in the immunohistochemical staining. The apoptotic cells increased significantly and the apoptotic index increased. The expression of MMP2,9 and eNOS in MMP2,9 and eNOS was increased by RT-PCR. (3) The blood flow velocity of the vein and the arterial segment increased, the blood pressure of the vein and the small amplitude of the arterial blood pressure were decreased after the operation of the arteriovenous system, the thickness of the intima of the arterial segment was increased, the thickness of the inner membrane of the vein was increased, and the content of the collagen fiber in the arterial segment was observed by the EVG staining. The content of collagen fibers in the vein segment was significantly increased, and the content of the elastic fibers was not statistically significant; the expression of PCNA in the arterial segment increased with the observation of the immunohistochemical staining, and the PCNA index of the venous segment increased significantly; the apoptotic cells observed in the arterial segment and the vein segment did not increase significantly after the apoptosis was stained. The expression of MMP2,9 and eON9 in the arterial segment was decreased, and the expression of MMP2,9 and eON9 in the venous segment was significantly reduced. Conclusion By establishing the model of arteriovenous graft and the model of arteriovenous malformation, the angle of the vascular remodeling is controlled by the change of blood flow dynamics, and the relevant theory is further improved. after the hemodynamic changes, the mechanical stress signal is conducted to the blood vessel wall cells, the expression of the enzymes (MMP2,9 and eONS, etc.) and the cytokines changes, the proliferation and apoptosis of the vascular wall cells and the change of the migration mechanism, the metabolic mechanisms of the extracellular matrix (collagen fibers and elastic fibers, etc.) are changed, Showing the change in the structure of the bleeding tube wall. The effect of the changes of hemodynamics on the vascular remodeling was further demonstrated, and the degree of the change of the hemodynamics was different, and the changes of the blood vessel remodeling were significantly different. At the same time, the theoretical hypothesis of the "Mechanical stress promotes proliferation and relieves mechanical stress and inhibits proliferation" is further verified in the course of blood vessel reconstruction.
【學(xué)位授予單位】：中國人民解放軍醫(yī)學(xué)院
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2015
【分類號(hào)】：R654.3
[Abstract]:The research background has shown that the mechanical stress is an important factor that affects the repair of the body, and the degree of repair can be reduced by reducing the mechanical stress. During the clinical work, the formation of the mechanical stress and the mechanical stress are closely related, the non-mechanical stress-free operation incision is not obvious, the slight mechanical stress at the end of the fracture leads to a large amount of hyperplasia of the bone, that is, the excessive repair, the non-mechanical stress can obtain the first-stage healing, No bone formation and so on, the fact that mechanical stress is an important factor that affects tissue repair. And many related basic studies have also confirmed this theory, can this theory be confirmed on the blood vessel, whether the change of hemodynamics can lead to the reconstruction of the vessel wall structure? It was found in the earlier work of the research group that:1. The uniform coating of a-cyanoacrylate adhesive on the blood vessel can achieve the purpose of fixing the blood vessel anastomosis. A large number of literature reported that the vascular wall of the vein had an arterial change after the vein graft was transplanted to the artery. In addition to that change in nutrition, the change in hemodynamics should be a major factor in the change in the tube wall. in addition, most of that current study focus on the study of the mechanism of vascular stenosis due to the change in the hemodynamic change of the clinically more common vein to the post-arterial blood flow, but less research on the regulation of the vascular remodeling mechanism for other hemodynamic changes, In this study, two models of the arteriovenous graft and the arteriovenous graft were used as the target, and the mechanism of the blood flow dynamic change and the regulation of the vascular remodeling was studied. Objective In order to further improve the theory of blood flow dynamics and control the mechanism of vascular remodeling, and to study the "Mechanical stress promotes proliferation and relieves mechanical stress and inhibits proliferation" of the skeletal muscle system, it is also applicable to the changes of blood flow dynamics to control the vascular remodeling. In this paper, we selected two models of the current study to study the correlation theory. The two models of the arteriovenous graft and the arteriovenous graft were used as the target, the parameters of the blood flow were measured, and the general, the tissue, the cell and the molecular biological level in the blood vessel reconstruction were observed and studied. And the relationship between the two elements is analyzed. Methods (1) The external diameter of the common carotid artery and the jugular vein and the femoral vein and the effective length of the femoral vein were measured respectively in the experimental animals and the blood vessel, and the experimental animals and the blood vessels established by the model were compared. (2) The establishment of the arteriovenous graft model, the regulation of the blood flow dynamics and the mechanism of the reconstruction of the blood vessel. The experimental animals were 12-15kg beagle dogs.1 cm of the left common carotid artery of the dog was cut and transplanted to the right external jugular vein, and the left common carotid artery and the right common carotid artery were ligated. The two groups of blood vessels were generally observed at 1,2 and 4 weeks after operation, and the thickness of the intimal hyperplasia was observed by HE staining and pathology. The changes of the content of the elastic fibers and the collagen fibers were observed by the histopathological observation, and the positive expression of the value-added cells was observed by immunohistochemical staining. The expression of MMP2,9 and eNOS gene was observed by RT-PCR. (3) The establishment of the arteriovenous blood vessel model, the regulation of the blood flow dynamics and the mechanism of the reconstruction of the blood vessel. The experimental animals were 12-15kg beagle dogs. The left common carotid artery and the external jugular vein of the dog were matched with the traditional needle-and-needle, and the right-hand common carotid artery and the external jugular vein were used as the control. The two groups of blood vessels were generally observed at 1,2 and 4 weeks after operation, and the thickness of the intimal hyperplasia was observed by HE staining and pathology. The changes of the content of the elastic fibers and the collagen fibers were observed by the histopathological observation, and the positive expression of the value-added cells was observed by immunohistochemical staining. The expression of MMP2,9 and eNOS gene was observed by RT-PCR. Results (1) The experimental animal beagle dog was more beneficial to the operation of the neck vessel in the SD rat, and the beagle dogs were more likely to measure the hemodynamic changes, and the beagle dogs were closer to the blood pressure and human blood pressure. (2) The blood flow velocity and blood pressure were significantly reduced after the artery was transplanted to the vein; the thickness of the intima was observed with HE staining; the content of the collagen fibers was significantly reduced by the EVG staining, and the content of the elastic fibers was not statistically significant; and the expression of PCNA was observed without significant change in the immunohistochemical staining. The apoptotic cells increased significantly and the apoptotic index increased. The expression of MMP2,9 and eNOS in MMP2,9 and eNOS was increased by RT-PCR. (3) The blood flow velocity of the vein and the arterial segment increased, the blood pressure of the vein and the small amplitude of the arterial blood pressure were decreased after the operation of the arteriovenous system, the thickness of the intima of the arterial segment was increased, the thickness of the inner membrane of the vein was increased, and the content of the collagen fiber in the arterial segment was observed by the EVG staining. The content of collagen fibers in the vein segment was significantly increased, and the content of the elastic fibers was not statistically significant; the expression of PCNA in the arterial segment increased with the observation of the immunohistochemical staining, and the PCNA index of the venous segment increased significantly; the apoptotic cells observed in the arterial segment and the vein segment did not increase significantly after the apoptosis was stained. The expression of MMP2,9 and eON9 in the arterial segment was decreased, and the expression of MMP2,9 and eON9 in the venous segment was significantly reduced. Conclusion By establishing the model of arteriovenous graft and the model of arteriovenous malformation, the angle of the vascular remodeling is controlled by the change of blood flow dynamics, and the relevant theory is further improved. after the hemodynamic changes, the mechanical stress signal is conducted to the blood vessel wall cells, the expression of the enzymes (MMP2,9 and eONS, etc.) and the cytokines changes, the proliferation and apoptosis of the vascular wall cells and the change of the migration mechanism, the metabolic mechanisms of the extracellular matrix (collagen fibers and elastic fibers, etc.) are changed, Showing the change in the structure of the bleeding tube wall. The effect of the changes of hemodynamics on the vascular remodeling was further demonstrated, and the degree of the change of the hemodynamics was different, and the changes of the blood vessel remodeling were significantly different. At the same time, the theoretical hypothesis of the "Mechanical stress promotes proliferation and relieves mechanical stress and inhibits proliferation" is further verified in the course of blood vessel reconstruction.
【學(xué)位授予單位】：中國人民解放軍醫(yī)學(xué)院
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2015
【分類號(hào)】：R654.3
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