本文選題：脊髓損傷(SCI) + 動(dòng)物模型��； 參考：《皖南醫(yī)學(xué)院》2015年碩士論文

【摘要】：目的:通過對(duì)隨機(jī)選入的SD大鼠采用環(huán)扎法制造大鼠脊髓損傷的模型,研究脊髓損傷后MP的使用與HSP70的表達(dá)變化規(guī)律,探究初期脊髓損傷的醫(yī)治策略,為進(jìn)一步學(xué)習(xí)脊髓損傷的模型打下堅(jiān)實(shí)的理論基礎(chǔ)。方法:本課題在前期工作基礎(chǔ)上再次通過大鼠環(huán)扎法造成大鼠脊髓損傷的模型,該方法具有很強(qiáng)的重復(fù)性,且方法簡(jiǎn)單穩(wěn)定,適合研究本課題。隨機(jī)選入成年SD雄性大鼠60只,大小均為13周齡,分為三組,第一組為空白組即A組:僅行椎板減壓,不作脊髓環(huán)扎處理。第二組為對(duì)照組即B組:采用環(huán)扎法進(jìn)行脊髓環(huán)扎,術(shù)后6小時(shí)作脊髓減壓處理;試驗(yàn)治療組為C組:同B組方法一致,也行脊髓環(huán)扎處理,在環(huán)扎法術(shù)后6小時(shí)減壓,但在環(huán)扎術(shù)后1小時(shí)給予大劑量MP靜脈注射治療。環(huán)扎術(shù)的關(guān)鍵就是充分地將研究段脊髓清晰暴露,本方法以蚊式血管鉗在肉眼情況下逐步咬除胸11椎板,暴露出脊髓,使用POLYSORB 5-0白色縫合線,精確地測(cè)量出硬膜囊周長(zhǎng)(C1),取出縫合線,通過公式(C2=C1×7.0)進(jìn)行運(yùn)算,得出需要的C2值,也就是硬膜囊原橫截面積的70%,通過可吸收線將原測(cè)量平面上的脊髓環(huán)扎至70%。之后分別于脊髓環(huán)扎術(shù)后第一天、第三天、第七天、第十四天和第二十一天等時(shí)間段對(duì)各組大鼠分批進(jìn)行處死,并取各組大鼠受損脊髓進(jìn)行制作病理切片,選用HE染色、免疫組化及原位末端脫氧核糖核酸轉(zhuǎn)移酶介導(dǎo)的脫氧尿苷三磷酸(dUTP)標(biāo)記法(TUNEL法)檢查和采用雙人盲法結(jié)合BBB評(píng)分、斜板法評(píng)價(jià)對(duì)各組結(jié)果進(jìn)行統(tǒng)計(jì)學(xué)分析,做出相關(guān)性研究。結(jié)果:空白組A組神經(jīng)運(yùn)動(dòng)功能BBB評(píng)分、斜板評(píng)分、HE染色均未見明顯異常改變;大鼠脊髓組織中HSP70陽性細(xì)胞數(shù)比較少見。與空白組A組相比,對(duì)照組B組大鼠的神經(jīng)運(yùn)動(dòng)功能BBB評(píng)分、斜板評(píng)分均降低,切片觀察脊髓組織中HSP70陽性細(xì)胞數(shù)于術(shù)后第1天開始升高,術(shù)后第3天達(dá)到最大值,差別有統(tǒng)計(jì)學(xué)意義(每組之間數(shù)據(jù)比較P0.05)。與對(duì)照組B組比較,治療組C組大鼠的神經(jīng)運(yùn)動(dòng)功能BBB評(píng)分、斜板評(píng)分明顯增高,脊髓組織中HSP70陽性細(xì)胞數(shù)明顯增加,HSP70陽性細(xì)胞數(shù)于術(shù)后第3天達(dá)到峰值,差異有統(tǒng)計(jì)學(xué)意義(各組之間數(shù)據(jù)比較P0.05)。結(jié)論:1、環(huán)扎法是通過縫合線對(duì)脊髓進(jìn)行持續(xù)性靜態(tài)壓迫而制作出大鼠急性脊髓損傷的模型,該方法的重復(fù)性良好,可定量化及可結(jié)合臨床,是一種比較可靠的脊髓損傷造模方法。2、實(shí)驗(yàn)組與對(duì)照組之間的BBB評(píng)分、斜板評(píng)分、HSP70陽性細(xì)胞數(shù)均存在統(tǒng)計(jì)學(xué)差異,由此可以推論MP可增加大鼠脊髓損傷后HSP70的表達(dá),而HSP70的作用機(jī)制為改善急性脊髓損傷后鈣離子的不平衡,抑制脂質(zhì)過氧化反應(yīng),避免自由基的損害,還能夠增加前列腺素的合成,改善損傷脊髓的血運(yùn)狀況,加強(qiáng)脊髓組織的修復(fù),并減少了脊髓損傷后繼發(fā)性損害的發(fā)生,起到了保護(hù)神經(jīng)的作用。3、脊髓損傷后,在外界因素的應(yīng)激作用下,機(jī)體內(nèi)HSP70表達(dá)增高,研究發(fā)現(xiàn)該蛋白為保護(hù)性蛋白,有保護(hù)機(jī)體脊髓神經(jīng)的功效,避免脊髓的進(jìn)一步損傷,對(duì)后期機(jī)體脊髓功能的恢復(fù)有一定的作用。
[Abstract]:Objective: To study the model of rat spinal cord injury by ligation of random selected SD rats, to study the changes in the use of MP and the expression of HSP70 after spinal cord injury, to explore the treatment strategy of spinal cord injury in the early stage, and to lay a solid theoretical basis for further learning of the model of spinal cord injury. The rat model of spinal cord injury caused by rat ring ligature was repeated, and the method was very repeatable, and the method was simple and stable. 60 adult SD male rats were randomly selected, the size of which was 13 weeks old and divided into three groups. The first group was the blank group, A group, only the vertebral lamina decompression and no spinal cord ligature. The second group was the same. Group B: group B: spinal cord ligature and decompression of spinal cord after operation. The test group was group C: same as group B, spinal cord cerclage, 6 hours after cerclage, 1 hours after cerclage, with large dose of MP intravenous injection. The key of cerclage was to fully study the spinal cord. In this method, the 11 vertebral lamina was gradually bitten by the mosquito type vascular forceps in the naked eye, the spinal cord was exposed, the POLYSORB 5-0 white suture was used, the perid sac circumference (C1) was accurately measured, the suture was taken out, the C2 value needed was obtained through the formula (C2=C1 x 7), that is, 70% of the original cross section of the dural sac, through the absorbable line. The spinal cord in the original measurement plane was ligation to 70%. after the first day, third days, seventh days, fourteenth days and twenty-first days after the spinal cord cerclage, and the injured spinal cord of each group was taken to make pathological sections, and HE staining, immunohistochemistry and in situ DDT were used. Transfer enzyme mediated deoxy uridine three phosphoric acid (dUTP) labelling (TUNEL) examination and double blind method combined with BBB score. The results of each group were statistically analyzed by oblique plate method, and the correlation study was made. Results: there was no obvious abnormal change in the BBB score of nerve movement function in the blank group A group, the score of oblique plate, HE staining, and the spinal cord tissue of rats. The number of HSP70 positive cells was rare. Compared with the A group in the blank group, the neuromotor function BBB score of the control group of the control group and the score of the oblique plate decreased. The number of HSP70 positive cells in the spinal cord tissue began to rise at first days after the operation, and the maximum value was reached on the third day after the operation, and the difference has the significance of the statistical significance (the comparison of the data between each group, P0.05). And the control. Group B was compared with the BBB score of the neuromotor function in the C group of the treatment group, the score of the oblique plate increased obviously, the number of HSP70 positive cells in the spinal cord increased obviously, and the number of HSP70 positive cells reached the peak at third days after the operation. The difference was statistically significant (the comparison of the data between each group was P0.05). Conclusion: the ring ligature method is the continuation of the spinal cord through the suture line to the spinal cord. The model of acute spinal cord injury in rats is made by sexual static compression. The reproducibility of this method is good. It can be quantified and can be combined with clinical. It is a reliable method of making model.2 for spinal cord injury. The BBB score, the score of oblique plate and the number of HSP70 positive cells between the experimental group and the control group are statistically different, thus the inference that the MP can be increased can be deduced. The expression of HSP70 after spinal cord injury in rats, and the action mechanism of HSP70 is to improve the imbalance of calcium ion after acute spinal cord injury, inhibit lipid peroxidation, avoid the damage of free radicals, increase the synthesis of prostaglandins, improve the blood transport status of the injured spinal cord, strengthen the repair of spinal cord tissue, and reduce the secondary sex of spinal cord injury. The occurrence of damage has played a role in protecting the nerve.3. After the injury of the spinal cord, the expression of HSP70 in the body is increased under the stress of external factors. The study found that the protein is a protective protein. It has the effect of protecting the spinal nerve of the body, avoiding the further injury of the spinal cord, and has a certain effect on the recovery of the spinal function of the later body.

【學(xué)位授予單位】：皖南醫(yī)學(xué)院
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2015
【分類號(hào)】：R651.2

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