【摘要】：目的:1、研制一種多功能小動物撞擊系統(tǒng)并評估其初步使用效果,為進(jìn)一步動物實(shí)驗(yàn)建立創(chuàng)傷模型及為滿足創(chuàng)傷基礎(chǔ)醫(yī)學(xué)研究提供儀器需要。2、建立一種新型的大鼠單純胰腺創(chuàng)傷模型,并基于損傷范圍評估其傷情變化,為后續(xù)研究提供模型條件。3、研究胰腺干細(xì)胞在本研究的大鼠胰腺大范圍創(chuàng)傷模型中增殖變化規(guī)律。方法:1、撞擊儀系統(tǒng)基于儲能換能裝置、簡便式多功能撞擊裝置、撞擊參數(shù)測量裝置等三大部件的相關(guān)原理設(shè)計,并進(jìn)一步檢驗(yàn)其有效性和穩(wěn)定性。2、將大鼠隨機(jī)分為2組,采用自主研發(fā)多功能撞擊儀的3cm2和6cm2撞擊頭,分別撞擊各組處于開腹?fàn)顟B(tài)的胰腺,使之在400kpa的壓強(qiáng)下形成對應(yīng)不等范圍的損傷,并設(shè)置對照;建模24h后觀察各組大鼠存活率、一般情況、大體病理和組織病理學(xué)改變,并檢測血鉀、血鈣的濃度和血清、腹水中淀粉酶、脂肪酶的水平。3、將大鼠隨機(jī)分為創(chuàng)傷組和對照組,前者接受6cm2/400Kpa的撞擊強(qiáng)度,后者為假手術(shù)組;建模后1、2、3、7天處死大鼠,處死前12h按100mg/kg體重經(jīng)腹腔注射5-溴-2'脫氧脲嘧啶核苷(Brd U);取胰腺組織,熒光TUNEL檢測細(xì)胞凋亡,Brd U的免疫組化染色檢測細(xì)胞增殖,RT-PCR、免疫組化(熒光)和Western blot等方法檢測PDX-1的表達(dá)并探討其變化規(guī)律。結(jié)果:1、多功能小動物撞擊系統(tǒng)研制完成,其撞擊應(yīng)力峰值連續(xù)可調(diào)節(jié)且量程在0-200kg,壓縮和擠壓應(yīng)力連續(xù)可調(diào)節(jié)且量程在0-100kg;經(jīng)驗(yàn)證,該儀器具有良好的有效性(P(27)0.05)和可重復(fù)性(P(29)0.05)。2、成功構(gòu)建了大鼠胰腺創(chuàng)傷模型;3cm2和6cm2致傷面積組均能形成明顯的損傷,其中致傷組死亡率、腹水量、胰腺濕重、病理評分、血清酶學(xué)指標(biāo)均高于對照組(P(27)0.05),血鉀、血鈣較對照組低(P(27)0.05),而6cm2致傷面積組的損傷比3cm2組更顯著。3、胰腺創(chuàng)傷模型中,傷后24h胰腺即可發(fā)生凋亡與代償性增殖;PDX-1的表達(dá)增強(qiáng)主要在建模后的第2-3天,隨后下降,至第7天恢復(fù)至正常水平附近,傷后第2,3天與同時間點(diǎn)對照組比較,有顯著性差異(P(27)0.05)。結(jié)論:1、研制的撞擊儀操作簡單,撞擊方式多樣,撞擊參數(shù)可被實(shí)時記錄;經(jīng)過調(diào)試和動物實(shí)驗(yàn),效果顯著且性能穩(wěn)定,可對不同實(shí)驗(yàn)動物形成不同強(qiáng)度和方式的創(chuàng)傷;適用于撞擊傷的建模和形態(tài)、機(jī)能學(xué)的研究。2、該胰腺創(chuàng)傷建模簡單、模型有效、操作的可重復(fù)性好,傷情單一且穩(wěn)定。適合用于傷情演化機(jī)制和創(chuàng)傷施救等研究。3、胰腺創(chuàng)傷后,胰腺干細(xì)胞將發(fā)生活化,在創(chuàng)傷后修復(fù)期增殖并分化為各類功能細(xì)胞,一定程度補(bǔ)償了胰腺受損害的功能。
[Abstract]:Objective: 1, to develop a multi-functional impact system for small animals and evaluate its preliminary effect, to establish trauma models for further animal experiments and to provide instruments for the basic medical research of trauma. A new rat model of simple pancreatic trauma was established, and the changes of injury were evaluated based on the range of injury, which provided the model condition for further study. 3, To investigate the proliferation of pancreatic stem cells in the rat pancreatic trauma model. Methods: 1. The impactor system was designed based on the principle of energy storage and energy exchanger, simple multi-function impact device and impact parameter measurement device, and further tested its effectiveness and stability. 2, the impact instrument system was designed based on the energy storage device, the convenient multi-function impact device and the impact parameter measurement device. The rats were randomly divided into two groups. The 3cm2 and 6cm2 impingement heads of the self-developed multi-function impingement instrument were used to impact the pancreas in the open state of each group respectively, so that the corresponding range of injury was formed under the pressure of 400kpa, and the control group was set up. The survival rate, general condition, gross pathological and histopathological changes of rats in each group were observed 24 hours after modeling, and the levels of serum potassium, serum calcium and amylase and lipase in serum and ascitic fluid were measured, and the levels of amylase and lipase in serum and ascitic fluid were measured. The rats were randomly divided into trauma group and control group, the former received the impact intensity of 6cm2/400Kpa, and the latter was sham-operated group. The rats were killed at 1, 2, 3, 7 days after modeling. The rats were injected intraperitoneally with 5-bromo-2'- deoxyuracil nucleoside (Brd U); 12 hours before death according to the body weight of 100mg/kg. The expression of PDX-1 in pancreatic tissue was detected by immunohistochemical staining of apoptotic, Brd U, and the expression of PDX-1 was detected by RT-PCR, immunohistochemistry (fluorescence) and Western blot, respectively. The expression of PDX-1 was detected by fluorescence TUNEL. Results: 1, the multi-function small animal impact system has been developed, the peak value of impact stress is continuously adjustable and the measuring range is 0 ~ 200 kg, the compression and extrusion stress is continuously adjustable and the measurement range is 0 ~ 100 kg; It was proved that the instrument was effective and reproducible (P (27) and repeatable (P (29 (0.05). (2) the rat pancreatic trauma model was successfully established. Both 3cm2 and 6cm2 injured area group could form obvious injury, in which death rate, ascites volume, wet weight of pancreas, pathological score, serum enzyme index were higher than those of control group (P (27), and serum potassium level was significantly higher than that of control group (P < 0.05). The level of serum calcium was lower than that of control group (P (27), but the injury of 6cm2 injury area group was more significant than that of 3cm2 group. 3. In pancreatic trauma model, 24 hours after injury, apoptosis and compensatory proliferation of pancreas could occur. The expression of PDX-1 increased mainly at the 2nd-3rd day after the model, then decreased, and returned to the normal level on the 7th day. There was a significant difference between the control group on the 2nd and 3rd day after injury and the control group at the same time (P (27). Conclusion: 1, the developed impactor is simple in operation, diverse in impact modes and can be recorded in real time, after debugging and animal experiments, the effect is remarkable and the performance is stable, which can cause trauma of different intensity and mode to different experimental animals; It is suitable for modeling and morphological, functional study of impact injury. 2, this pancreatic trauma model is simple, effective, repeatable, simple and stable. (3) Pancreatic stem cells will be activated after pancreatic trauma, proliferate and differentiate into various functional cells in post-traumatic period, which can compensate the damaged function of pancreas to a certain extent. 3) it can be used to study the mechanism of wound evolution and wound rescue. 3) after pancreatic trauma, pancreatic stem cells will be activated and differentiated into various functional cells.
【學(xué)位授予單位】：西南醫(yī)科大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2017
【分類號】：R657.5;R-332

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