本文選題：HSP70 + 閉合性腦損傷��； 參考：《南昌大學(xué)》2010年碩士論文

【摘要】： 目的:探索HSP70蛋白及其mRNA在大鼠不同等級(jí)閉合性腦損傷中的表達(dá);探索大鼠閉合性腦損傷分級(jí)自由落體打擊技術(shù);探討HSP70在大鼠不同等級(jí)閉合性腦損傷中的意義。 方法:1、根據(jù)自由落體原理,采用不同重量的砝碼從同一高度下落,以不同沖擊應(yīng)力(128.90Kpa、386.70Kpa、644.50Kpa)的方法,造成輕、中、重度大鼠閉合性腦損傷模型共54只為實(shí)驗(yàn)組,顱腦假手術(shù)18只大鼠為對(duì)照組。2、應(yīng)用免疫組織化學(xué)EliVision方法,檢測(cè)不同等級(jí)大鼠閉合性腦損傷中HSP70蛋白的表達(dá)變化情況。3、采用逆轉(zhuǎn)錄多聚酶鏈反應(yīng)(RT-PCR)技術(shù)對(duì)不同等級(jí)大鼠閉合性腦損傷中HSP70mRNA進(jìn)行定量分析。 結(jié)果:1、HE染色結(jié)果顯示傷后3h輕度損傷組軟腦膜下皮質(zhì)出血,神經(jīng)膠質(zhì)細(xì)胞增生,中度損傷組皮質(zhì)淺層出血,腦實(shí)質(zhì)水腫,神經(jīng)膠質(zhì)細(xì)胞增生,重度損傷組皮質(zhì)廣泛出血,出血灶周圍見中性粒細(xì)胞浸潤(rùn),神經(jīng)元消失,周邊腦實(shí)質(zhì)網(wǎng)格樣。2、免疫組織化學(xué)染色結(jié)果顯示傷后3h HSP70蛋白在對(duì)照組、輕度損傷組、中度損傷組及重度損傷組均有不同程度的表達(dá),其陽(yáng)性表達(dá)率分別為8.1%、11.24%、20.56%、30.43%;輕度損傷組與對(duì)照組之間表達(dá)差異無(wú)統(tǒng)計(jì)學(xué)意義(P0.05);中度、重度損傷組與對(duì)照組表達(dá)差異有統(tǒng)計(jì)學(xué)意義(P0.05);輕、中、重度損傷組之間表達(dá)差異有統(tǒng)計(jì)學(xué)意義(P0.05)。3、RT-PCR結(jié)果顯示傷后3 h HSP70mRNA在對(duì)照組、輕度損傷組、中度損傷組及重度損傷組均有不同程度的表達(dá),HSP70與β-actin的光密度比值平均值分別為0.330、0.415、0.550、0.767;輕度損傷組與對(duì)照組之間比值差異無(wú)統(tǒng)計(jì)學(xué)意義(P0.05);中度、重度損傷組與對(duì)照組比值差異有統(tǒng)計(jì)學(xué)意義(P0.05);輕、中、重度損傷組之間比值差異有統(tǒng)計(jì)學(xué)意義(P0.05)。 結(jié)論: 1、HSP70mRNA及蛋白在應(yīng)激狀態(tài)下的神經(jīng)元損傷的表達(dá)是一致的,表明短時(shí)間內(nèi)HSP70mRNA及其蛋白表達(dá)率可隨著損傷程度的加重明顯升高,HSP70的表達(dá)差異可作為短時(shí)間內(nèi)中度以上閉合性腦損傷程度的一個(gè)較好的分級(jí)指標(biāo)。2、該模型解決并完善了經(jīng)典落體打擊法中打擊部位不穩(wěn)定的問題,重復(fù)性好,對(duì)閉合性腦損傷實(shí)驗(yàn)研究有實(shí)用價(jià)值。
[Abstract]:Objective: to investigate the expression of HSP70 protein and its mRNA in rat brain injury of different grades, to explore the technique of free falling body attack in different grades of closed brain injury in rats, and to explore the significance of HSP70 in different grades of closed brain injury in rats. Methods according to the principle of free falling body, 54 rats with light, moderate and severe closed brain injury were induced by using different weights falling from the same height and different impact stress (128.90Kpa-386.70Kpa-644.50Kpaa). Eighteen rats with craniocerebral sham-operation were used as control group (n = 18). Immunohistochemical EliVision method was used. The changes of HSP70 protein expression in closed brain injury of rats of different grades were detected. Reverse transcriptase polymerase chain reaction (RT PCR) technique was used to quantitatively analyze HSP70mRNA in closed brain injury of rats of different grades. Results the results showed that the subpial cortex hemorrhage, glial cell proliferation, superficial cortical hemorrhage, cerebral parenchyma edema, glial cell proliferation and extensive hemorrhage in severe injury group were found in mild injury group 3 hours after injury. Neutrophil infiltration and neuronal disappearance were observed around the hemorrhage foci. The results of immunohistochemical staining showed that HSP70 protein was present in the control group and mild injury group at 3 h after injury. The positive expression rates of moderate injury group and severe injury group were 8.1and 11.240.56and 30.433.There was no significant difference between mild injury group and control group (P 0.05). There was significant difference in the expression of HSP70mRNA between the severe injury group and the control group (P 0.05), and between the mild, moderate and severe injury groups, the RT-PCR results showed that the expression of HSP70mRNA was in the control group and the mild injury group at 3 hours after injury. The average optical density ratio of HSP70 and 尾 -actin in moderate injury group and severe injury group were 0.330, 0.415, 0.550 and 0.767.There was no significant difference between mild injury group and control group (P 0.05). There was significant difference in the ratio between severe injury group and control group (P 0.05), while there was significant difference between light, moderate and severe injury group (P 0.05). Conclusion: 1 the expression of HSP70 mRNA and protein in the injured neurons under stress is consistent. The results indicate that the expression rate of HSP70mRNA and its protein can increase significantly with the severity of injury in a short period of time, and the difference of HSP70 expression can be used as a good grading index of the degree of closed brain injury in a short period of time. Improved the problem of unstable hit parts in the classical falling body attack method, It has good reproducibility and has practical value for the experimental study of closed brain injury.
【學(xué)位授予單位】：南昌大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2010
【分類號(hào)】：R363

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