本文選題：Kr(u|)ppel樣轉(zhuǎn)錄因子-2 + 高遷移率族蛋白-B1　； 參考：《昆明醫(yī)科大學(xué)》2015年碩士論文

【摘要】：深靜脈血栓因其形成原因復(fù)雜多變,應(yīng)用傳統(tǒng)診斷方法難以進(jìn)行早期確診,早期確診困難的現(xiàn)狀嚴(yán)重影響了治療的及時(shí)性,如不能得到及時(shí)有效的早期診治,可引發(fā)肢體進(jìn)一步病理性損害,并可能會(huì)遺留肢體靜脈功能不全；嚴(yán)重者血栓脫落隨血液循環(huán)進(jìn)入肺組織引起肺動(dòng)脈栓塞,甚至可危及生命。因此,與DVT早期診斷和預(yù)防相關(guān)的標(biāo)志物的研究一直是國(guó)內(nèi)外研究者的探討熱點(diǎn)。近年來(lái),眾多可引起DVT的復(fù)雜因素中,圍繞內(nèi)皮細(xì)胞損傷方面的討論逐漸成為DVT發(fā)病原因的研究熱點(diǎn)。內(nèi)皮細(xì)胞損傷釋放出的各種炎癥因子可激活白細(xì)胞,活化血小板,導(dǎo)致或加速了血栓的形成。KLF-2、HMGB-1與DVT形成的關(guān)鍵角色內(nèi)皮細(xì)胞損傷關(guān)系密切,通過(guò)保護(hù)內(nèi)皮細(xì)胞膜穩(wěn)定,負(fù)向調(diào)控炎癥因子等功能,發(fā)揮了抑制血栓的作用。本研究圍繞KLF-2、HMGB-1進(jìn)行研究,實(shí)驗(yàn)通過(guò)DVT動(dòng)物模型構(gòu)建,結(jié)合分子生物學(xué)技術(shù)檢測(cè)手段,觀察上述標(biāo)志物在DVT形成過(guò)程中的表達(dá)變化,初步討論兩種細(xì)胞因子的內(nèi)在聯(lián)系及其在深靜脈血栓形成中的作用。目的1.下腔靜脈狹窄法建立C57小鼠DVT模型,病理切片觀察血栓形成情況,獲取各組小鼠下腔靜脈組織；2. Real-time PCR檢測(cè)下腔靜脈和血栓組織中,KLF-2、HMGB-1在轉(zhuǎn)錄層面的表達(dá)變化,初步探討其與DVT的聯(lián)系及作用,為進(jìn)一步分析其與DVT的關(guān)系提供基礎(chǔ)實(shí)驗(yàn)證據(jù)；3. ELISA法檢測(cè)血漿中t-PA蛋白的表達(dá)變化,探討其與DVT的聯(lián)系及作用,為進(jìn)一步分析其與DVT的關(guān)系提供基礎(chǔ)實(shí)驗(yàn)證據(jù)。方法1.建立DVT小鼠模型并取材行相關(guān)檢測(cè)：將120只C57小鼠采用隨機(jī)分組原則,分為正常對(duì)照組(n=40)、假手術(shù)(n=40)和DVT組(n=40)。造模后24小時(shí),正常組、假手術(shù)組取腎靜脈下方1.5cm下腔靜脈組織；DVT組取血栓形成范圍內(nèi)下腔靜脈組織及其內(nèi)容物；并記錄下腔靜脈直徑等數(shù)據(jù)。2.下腔靜脈組織樣本行病理切片,HE染色后觀察血栓形成情況；Real-time PCR檢測(cè)各組靜脈組織中KLF-2、HMGB-1基因mRNA表達(dá)水平,分析其與DVT形成關(guān)系。3. ELISA法檢測(cè)各組血漿中t-PA蛋白的表達(dá)變化；分析其與DVT形成關(guān)系。結(jié)果1.DVT組小鼠模型靜脈組織HE染色：DVT組可見(jiàn)完全血栓形成、血管壁內(nèi)炎細(xì)胞侵潤(rùn)明顯；正常組、假手術(shù)組未見(jiàn)形成血栓。2. Real-time PCR檢測(cè)靜脈組織KLF-2 mRNA表達(dá)：DVT組較其它兩組表達(dá)上調(diào),具統(tǒng)計(jì)學(xué)意義(P0.05)；正常對(duì)照組與假手術(shù)組相比(P0.05),沒(méi)有統(tǒng)計(jì)學(xué)差異。HMGB-1 mRNA表達(dá)：DVT組較正常對(duì)照組及假手術(shù)組表達(dá)下調(diào),具統(tǒng)計(jì)學(xué)意義(P0.05)；正常對(duì)照組與假手術(shù)組相比(P0.05),沒(méi)有統(tǒng)計(jì)學(xué)差異。3. ELISA法檢測(cè)血漿中t-PA蛋白表達(dá)：DVT組較正常對(duì)照組及假手術(shù)組表達(dá)上調(diào),具統(tǒng)計(jì)學(xué)意義(P0.05)；正常對(duì)照組與假手術(shù)組相比(P0.05),沒(méi)有統(tǒng)計(jì)學(xué)差異。結(jié)論1.小鼠DVT模型中KLF-2、HMGB-1、t-PA表達(dá)變化提示,它們與DVT形成有關(guān)；在DVT形成初期它們對(duì)血栓形成可能起到一定作用。2. KLF-2、HMGB-1可能通過(guò)影響t-PA蛋白含量,從而打破纖溶-抗纖系統(tǒng)動(dòng)態(tài)平衡,起到影響血栓形成的作用。
[Abstract]:The cause of deep venous thrombosis is complicated and changeable, and it is difficult to make early diagnosis with traditional diagnostic method. The difficult situation of early diagnosis seriously affects the timeliness of treatment. If it can't get timely and effective diagnosis and treatment, it can cause further pathological damage of limbs, and can leave limb venous insufficiency; serious thrombosis The study of the markers associated with early diagnosis and prevention of DVT has been a hot spot for researchers both at home and abroad. In recent years, many of the complex factors that can cause DVT have gradually become the cause of the pathogenesis of DVT. The various inflammatory factors released by endothelial cell damage can activate leukocytes, activate platelets, lead to or accelerate the formation of.KLF-2, and HMGB-1 is closely related to the key role of endothelial cells in the formation of DVT, and the inhibition of thrombus is played by protecting endothelial cell membrane stability and negatively regulating inflammatory factors. This study was conducted around KLF-2 and HMGB-1. The experiment was constructed by DVT animal model and combined with molecular biological techniques to observe the changes in the expression of the above markers in the formation of DVT. The internal relations of the two cytokines and their role in the formation of deep venous thrombosis were preliminarily discussed. Objective 1. inferior vena cava stenosis method was established. C57 mice DVT model, pathological sections to observe the formation of thrombosis, to obtain the inferior vena cava tissue in each group; 2. Real-time PCR detection of the inferior vena cava and thrombus tissue, KLF-2, HMGB-1 at the transcriptional level changes, preliminarily explore its relationship with DVT and the role of the further analysis of the relationship with DVT to provide basic experimental evidence; 3. EL ISA method was used to detect the changes in the expression of t-PA protein in plasma and to explore its relationship with DVT and to provide basic experimental evidence for further analysis of its relationship with DVT. Method 1. the model of DVT mice was established and the correlation detection was taken. 120 C57 mice were randomly divided into normal control group (n=40), sham operation (n=40) and DVT group (n=40). 24 hours after the model, the normal group and the sham group took the inferior vena cava inferior vena cava below the renal vein in the sham operation group; the DVT group took the inferior vena cava tissue and its contents within the range of thrombus formation, and recorded the lower vena cava diameter and other data of the inferior vena cava tissue in.2., and observed the formation of thrombus after HE staining; Real-time PCR was used to detect the veins in.2.. The expression level of mRNA, HMGB-1 gene mRNA in tissue, and analysis of the relationship between the expression of t-PA protein in the plasma with.3. ELISA method and DVT formation, and the relationship with DVT formation. Results the 1.DVT group mice model venous tissue HE staining: DVT group can see complete thrombosis, inflammatory cells in the vascular wall are invaded obviously; normal group, sham operation group. The expression of KLF-2 mRNA in venous tissue was not detected by the formation of thrombus.2. Real-time PCR: the expression of DVT in the group of the other two groups was up, with statistical significance (P0.05); there was no statistical difference of.HMGB-1 mRNA expression between the normal control group and the sham operation group (P0.05), and the expression of the DVT group was lower than that of the normal control group and the sham operation group, with a statistical significance (P0.05). Compared with the sham operation group (P0.05), there was no statistical difference between the normal control group and the sham group (P0.05) to detect the expression of t-PA protein in the plasma: the expression of t-PA in the DVT group was higher than that in the normal control group and the sham operation group (P0.05), and there was no statistical difference between the normal control group and the sham operation group (P0.05). Conclusion the 1. mice DVT model was found to be in KLF-2 and HMGB-1, The changes in the expression of t-PA suggest that they are related to the formation of DVT; they may play a role in the formation of.2. KLF-2 in the early stage of the formation of DVT, and HMGB-1 may affect the dynamic balance of fibrinolytic and fibrinolytic system by affecting the content of t-PA protein, thus affecting the formation of thrombus.
【學(xué)位授予單位】：昆明醫(yī)科大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2015
【分類(lèi)號(hào)】：R543.6

【參考文獻(xiàn)】

相關(guān)期刊論文 前1條

1 宮玉玲;邢啟崇;;高遷移率族蛋白B1與動(dòng)脈粥樣硬化[J];中國(guó)動(dòng)脈硬化雜志;2008年09期

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本文編號(hào)：1898176