【摘要】：目的：高壓電燒傷是一種特殊原因的燒傷，可引起一系列復(fù)雜的病理學(xué)變化，微循環(huán)障礙則是其中重要的病理變化之一。凝血機(jī)制異常在機(jī)體微循環(huán)障礙發(fā)生和發(fā)展過程中起重要作用。燒傷后由于血管內(nèi)皮受損、血小板活化、炎癥介質(zhì)釋放、凝血系統(tǒng)功能增強(qiáng)及繼發(fā)性纖溶系統(tǒng)功能相對(duì)增強(qiáng)，導(dǎo)致血液高凝狀態(tài)，甚至引起彌漫性血管內(nèi)凝血（disseminatedintravascular coagulation，DIC）。血漿纖維蛋白原（fibrinogen，F(xiàn)IB）、D-二聚體是血液發(fā)生血栓及血栓前狀態(tài)的凝血及纖溶系統(tǒng)活性改變的分子標(biāo)志物，可反映體內(nèi)凝血和纖溶過程的變化；纖維連接蛋白（fibronectin，F(xiàn)N）在血小板聚集和黏附過程中起重要作用，可反映燒傷后血液的黏滯性。因此，我們設(shè)計(jì)本實(shí)驗(yàn)，旨在研究高壓電燒傷早期大鼠FIB、D-二聚體、FN變化，并通過烏司他�。╱linastain，UTI）進(jìn)行干預(yù)，研究燒傷后凝血功能和纖溶活性的分子機(jī)制，探討UTI治療高壓電燒傷的可行性。 方法： 1.實(shí)驗(yàn)動(dòng)物分組：健康成年SD大鼠（由河北醫(yī)科大學(xué)動(dòng)物實(shí)驗(yàn)中心提供，合格證編號(hào)為1211057）180只，按隨機(jī)數(shù)字表法分成三組，即假高壓電燒傷組（簡稱對(duì)照組）、高壓電燒傷組（簡稱電傷組）、高壓電燒傷UTI治療組（簡稱治療組），每組各60只。每組又按觀察時(shí)間分為電擊前15min、電擊后5min、電擊后1h、2h、4h、8h六個(gè)時(shí)相，即T0、T1、T2、T3、T4、T5，每時(shí)相10只。 2.實(shí)驗(yàn)前準(zhǔn)備：將大鼠編號(hào)、稱重，左上肢、右下肢及前胸脫毛。將實(shí)驗(yàn)藥品按實(shí)驗(yàn)需要配成所需濃度。 3.高壓電燒傷模型制作：首先連接好實(shí)驗(yàn)變壓器和調(diào)壓器電線。用1%戊巴比妥鈉腹腔注射麻醉大鼠，按40mg/kg給藥，麻醉成功后，將大鼠仰臥于專用電擊實(shí)驗(yàn)臺(tái)上，固定四肢，將兩個(gè)1cm×1cm電極片分別固定于大鼠的左上肢（電流入口）、右下肢（電流出口）脫毛區(qū)。接通電源后，調(diào)整調(diào)壓器使升壓器輸出電壓至2kV，連接升壓器，使高壓電流通過大鼠，電擊3s。對(duì)照組制作假電傷模型，不合閘通過電流，其余步驟與電傷組一致。電擊傷后即刻，治療組腹腔注射5×104u/kg UTI，對(duì)照組及電傷組腹腔內(nèi)注射等量的生理鹽水。 4.標(biāo)本采集與保存：將高壓電燒傷模型復(fù)制成功的大鼠開胸暴露心臟，直視下心臟抽血6mL，其中3mL置于一次性使用真空采血管（紅）中，3mL置于一次性使用真空采血管（藍(lán)）中，輕輕顛倒數(shù)次后靜置30min，上離心機(jī)，以3000轉(zhuǎn)/min離心10min，紅管中取上清液置于Eppendorf管中在-70℃條件下保存。 5．指標(biāo)檢測：藍(lán)管離心分離血漿后使用全自動(dòng)血凝儀上機(jī)自動(dòng)檢測每組大鼠六個(gè)時(shí)相組FIB及D-二聚體含量。置于冰箱的血清采用ELISA雙抗體夾心法，檢測FN含量。 6．實(shí)驗(yàn)數(shù)據(jù)處理：采用SPSS13.0統(tǒng)計(jì)軟件，行兩因素析因設(shè)計(jì)的方差分析，多重比較采用LSD法t檢驗(yàn)。以P＜0.05為有統(tǒng)計(jì)學(xué)差異。 結(jié)果： 1．大鼠血漿FIB含量變化 電傷組FIB含量總體高于對(duì)照組（P＜0.01），且FIB含量受傷后受時(shí)間因素的影響（P＜0.01）,至傷后8h達(dá)到最高值。治療組FIB含量總體低于電傷組（P＜0.01），且FIB含量治療后受時(shí)間因素的影響（P＜0.01），，但只有治療組傷后5min高于本組傷前值（P＜0.01），傷后1h、2h、4h、8h與傷前無明顯差異（P＞0.05）。 2．大鼠血漿D-二聚體含量變化 電傷組D-二聚體含量總體高于對(duì)照組（P＜0.01），且D-二聚體含量受傷后受時(shí)間因素的影響（P＜0.01），呈增高趨勢。治療組D-二聚體含量總體低于電傷組（P＜0.01），且D-二聚體含量治療后受時(shí)間因素的影響（P＜0.01），治療組傷后5min～8h各時(shí)相均高于本組傷前值（P＜0.001） 3．大鼠血清FN含量變化 電傷組FN含量總體低于對(duì)照組（P＜0.01），且FN含量受傷后受時(shí)間因素的影響（P＜0.05），傷后各時(shí)相均低于本組傷前值（P＜0.05）。治療組FN含量總體高于電傷組（P＜0.05），但FN含量治療后不受時(shí)間因素的影響（P＞0.05）。 結(jié)論： 1．高壓電燒傷早期大鼠血漿FIB、D-二聚體表達(dá)水平均有不同程度升高，血清FN表達(dá)水平下降，說明高壓電燒傷后存在高凝狀態(tài)、纖溶系統(tǒng)功能異常及微循環(huán)障礙。 2．高壓電燒傷早期UTI治療后FIB、D-二聚體表達(dá)水平降低，F(xiàn)N表達(dá)水平升高，說明UTI治療可以抑制凝血亢進(jìn)與繼發(fā)性纖溶，減輕高凝狀態(tài)，從而改善微循環(huán)，防止血栓形成。
[Abstract]:Objective: High voltage electrical burn is a special cause of burns, which can cause a series of complex pathological changes. Microcirculation disorder is one of the important pathological changes. Abnormal coagulation mechanism plays an important role in the occurrence and development of microcirculation disorder. Release, enhancement of coagulation system and secondary fibrinolysis system lead to hypercoagulability, even diffuse intravascular coagulation (DIC). Plasma fibrinogen (FIB) and D-dimer are coagulation and fibrinolysis system activities in the thrombosis and pre-thrombotic state of the blood. The changes of molecular markers can reflect the changes of coagulation and fibrinolysis in vivo. Fibronectin (FN) plays an important role in the process of platelet aggregation and adhesion, and can reflect the blood viscosity after burns. Ulinastatin (UTI) was used to study the molecular mechanism of coagulation function and fibrinolytic activity after burns, and to explore the feasibility of UTI in the treatment of high voltage electrical burn.
Method:
1. Experimental animal groups: 180 healthy adult SD rats (provided by the Animal Experimental Center of Hebei Medical University, with the certificate number of 121,1057) were randomly divided into three groups: sham high-voltage electrical burn group (control group), high-voltage electrical burn group (electrical burn group), high-voltage electrical burn UTI treatment group (treatment group), 60 rats in each group. Group A was divided into six phases, namely, T0, T1, T2, T3, T4, T5, 5 minutes before and 1, 2, 4 and 8 hours after electric shock.
2. Pre-experiment preparation: Rats were numbered, weighed, left upper extremity, right lower extremity and chest depilation.
3. Making high voltage electric burn model: Firstly, the experimental transformer and the voltage regulator wire were connected. Anesthetized rats were injected intraperitoneally with 1% sodium pentobarbital. After anesthesia, the rats were given 40 mg/kg. After successful anesthesia, the rats were lying on their backs on a special electric shock test bench, fixed their limbs, and fixed two 1 cm *1 cm electrode plates on the left upper limbs (current inlet) and right lower limbs (current inlet). After switching on the power supply, adjust the voltage regulator to make the output voltage of the booster to 2 kV, and connect the booster to make the high voltage current pass through the rats and shock them for 3 s. The control group made the model of false electric injury, which was not suitable for the switch through the current, and the other steps were the same as that of the electric injury group. The same amount of saline was injected into the abdominal cavity.
4. Sample Collection and Preservation: The heart was exposed after thoracotomy in rats with high-voltage electrical burn. Blood was drawn from the heart under direct vision for 6 mL. Among them, 3 mL was placed in a disposable vacuum (red), 3 mL was placed in a disposable vacuum (blue), and 30 minutes was left after several times of inversion. The heart was centrifuged for 10 minutes at 3000 rpm. The supernatant was placed in the Eppendorf tube and stored at -70 C.
5. Indicators: After centrifugation, the plasma was separated by blue tube and the contents of FIB and D-dimer were detected automatically by automatic coagulator. The serum in refrigerator was detected by ELISA double antibody sandwich method.
6. Data processing: SPSS13.0 statistical software was used to analyze the variance of two-factor factorial design, and LSD t test was used for multiple comparisons.
Result:
Changes of plasma FIB content in 1. rats
The content of FIB in the treatment group was lower than that in the electric injury group (P < 0.01), but only 5 minutes after injury was higher than that in the control group (P < 0.01). There was no significant difference in 1H, 2h, 4H and 8h after injury (P > 0.05).
Changes of plasma D- two dimer content in 2. rats
The content of D-dimer in the treatment group was lower than that in the electric injury group (P < 0.01), and the content of D-dimer in the treatment group was higher than that in the control group (P The pre injury values were (P < 0.001).
Changes of serum FN level in 3. rats
The FN content in the treatment group was higher than that in the electric injury group (P < 0.05), but the FN content was not affected by the time factor (P > 0.05).
Conclusion:
1. The levels of FIB and D-dimer in plasma and FN in serum of rats with high-voltage electrical burn increased to different degrees in the early stage, indicating that there was hypercoagulable state, abnormal fibrinolytic system function and microcirculation disturbance after high-voltage electrical burn.
2. After early UTI treatment, the expression of FIB, D-dimer and FN decreased, indicating that UTI treatment can inhibit hypercoagulation and secondary fibrinolysis, reduce hypercoagulability, improve microcirculation and prevent thrombosis.
【學(xué)位授予單位】：河北醫(yī)科大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2013
【分類號(hào)】：R647

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