本文選題：創(chuàng)傷性顱腦損傷 + 丙戊酸��； 參考：《新鄉(xiāng)醫(yī)學(xué)院》2013年碩士論文

【摘要】：背景 創(chuàng)傷性顱腦損傷(Traumatic Brain Injury, TBI)是神經(jīng)外科常見(jiàn)急癥,TBI后由于神經(jīng)細(xì)胞的凋亡、壞死及炎癥反應(yīng)等繼發(fā)性損傷,造成運(yùn)動(dòng)和感覺(jué)功能障礙,嚴(yán)重影響患者預(yù)后。目前,對(duì)神經(jīng)細(xì)胞的繼發(fā)損傷尚無(wú)切實(shí)有效的治療方法。因此TBI后早期進(jìn)行神經(jīng)細(xì)胞的保護(hù)及炎癥反應(yīng)的控制是神經(jīng)外科研究的一個(gè)熱點(diǎn)。近年的研究發(fā)現(xiàn)丙戊酸(valproic acid, VPA)具有多方面的神經(jīng)營(yíng)養(yǎng)和神經(jīng)保護(hù)作用。本研究在建立閉合性顱腦損傷動(dòng)物模型的基礎(chǔ)上,早期應(yīng)用丙戊酸,通過(guò)行為學(xué)評(píng)分和腦組織內(nèi)神經(jīng)細(xì)胞凋亡情況、HSP70和caspase-3、TNF-α和IL-1β蛋白表達(dá)的變化,探討VPA對(duì)TBI大鼠神經(jīng)功能恢復(fù)的影響及其機(jī)制,為臨床應(yīng)用提供理論基礎(chǔ)。 目的 通過(guò)建立大鼠閉合性顱腦損傷模型,觀察丙戊酸干預(yù)后,不同時(shí)間點(diǎn)大鼠腦組織內(nèi)神經(jīng)細(xì)胞凋亡情況、HSP70和caspase-3、TNF-α和IL-1β蛋白表達(dá)的影響,及腦組織中TNF-α和IL-1β的動(dòng)態(tài)變化,探討丙戊酸對(duì)實(shí)驗(yàn)性顱腦損傷大鼠的抗細(xì)胞凋亡、神經(jīng)保護(hù)作用和減輕炎性反應(yīng)的作用。 方法 將健康SD雄性大鼠隨機(jī)分成正常對(duì)照組、單純損傷組、VPA治療組。建立大鼠閉合性顱腦損傷模型,采用神經(jīng)系統(tǒng)疾病嚴(yán)重程度評(píng)分(Neurological Severity Scores, NSS)來(lái)評(píng)價(jià)各組大鼠不同時(shí)間點(diǎn)的神經(jīng)功能；運(yùn)用光學(xué)顯微鏡觀察腦組織形態(tài)學(xué)的改變,應(yīng)用原位末端轉(zhuǎn)移酶標(biāo)記技術(shù)(TUNEL)檢測(cè)大鼠腦組織內(nèi)細(xì)胞凋亡情況；運(yùn)用免疫組織化學(xué)染色法(SP法)對(duì)各組不同時(shí)間點(diǎn)內(nèi)腦細(xì)胞中的HSP70和caspase-3蛋白進(jìn)行檢測(cè)；利用免疫熒光技術(shù)檢測(cè)各組不同時(shí)間點(diǎn)內(nèi)腦細(xì)胞中的TNF-α和IL-1β蛋白表達(dá)情況；采用酶聯(lián)免疫吸附試驗(yàn)(ELISA)法測(cè)定各組不同時(shí)間點(diǎn)內(nèi)腦組織中TNF-α和IL-1β的動(dòng)態(tài)變化,并作統(tǒng)計(jì)學(xué)分析。 結(jié)果 1.行為學(xué)測(cè)定顯示：大鼠行為學(xué)評(píng)分結(jié)果VPA治療后大鼠神經(jīng)功能有不同程度的恢復(fù),在創(chuàng)傷后3d內(nèi)3組間差異無(wú)顯著性意義(P0.05); VPA治療后7d、14d、21d、28d, VPA治療組神經(jīng)功能恢復(fù)程度優(yōu)于單純損傷組,兩組間比較差異有顯著性意義(P0.05)。 2.原位末端標(biāo)記法(TUNEL)檢測(cè)顯示：?jiǎn)渭儞p傷組與VPA治療組大鼠損傷區(qū)皮質(zhì)各時(shí)間點(diǎn)凋亡陽(yáng)性細(xì)胞數(shù)均高于正常對(duì)照組,差異均有統(tǒng)計(jì)學(xué)意義(P0.05)。VPA治療組各時(shí)間點(diǎn)凋亡陽(yáng)性細(xì)胞數(shù)均低于單純損傷組,差異均有統(tǒng)計(jì)學(xué)意義(P0.05)。 3.免疫組織化學(xué)檢測(cè)顯示：VPA治療組相同時(shí)間點(diǎn)HSP70平均灰度值明顯低于單純損傷組,兩者差異有統(tǒng)計(jì)學(xué)意義(P0.05)。VPA治療組相同時(shí)間點(diǎn)caspase-3平均灰度值明顯高于單純損傷組,兩者差異有統(tǒng)計(jì)學(xué)意義(P0.05)。 4.免疫熒光技術(shù)檢測(cè)：VPA治療組相同時(shí)間點(diǎn)腦組織TNF-α和IL-1β陽(yáng)性細(xì)胞數(shù)明顯低于單純損傷組,兩者差異有統(tǒng)計(jì)學(xué)意義(P0.05)。 5.酶聯(lián)免疫吸附試驗(yàn)(ELISA)檢測(cè)：造模后單純損傷組和VPA治療組相同時(shí)間點(diǎn)腦組織中TNF-α和IL-1β蛋白濃度均高于正常對(duì)照組,差異均有統(tǒng)計(jì)學(xué)意義(P0.05); VPA治療組相同時(shí)間點(diǎn)腦組織中TNF-α和IL-1β蛋白濃度均低于單純損傷組,差異均有統(tǒng)計(jì)學(xué)意義(P0.05)。 結(jié)論 VPA治療后能夠改善創(chuàng)傷性顱腦損傷大鼠的神經(jīng)功能；VPA有抗細(xì)胞凋亡、保護(hù)神經(jīng)細(xì)胞和減輕炎性反應(yīng)的作用。
[Abstract]:background
Traumatic brain injury (Traumatic Brain Injury, TBI) is a common emergency in the Department of neurosurgery. After TBI, the secondary injury of nerve cell apoptosis, necrosis and inflammatory reaction causes motor and sensory dysfunction, which seriously affects the prognosis of the patients. At present, there is no effective and effective treatment for the injury of nerve cells. So TBI early after the injury. The protection of the nerve cells and the control of the inflammatory reaction are a hot spot in the Department of neurosurgery. In recent years, the study found that valproic acid (VPA) has many aspects of neurotrophic and neuroprotective effects. The changes of neuronal apoptosis and the expression of HSP70 and Caspase-3, TNF- alpha and IL-1 beta protein in the brain tissue, and to explore the effect and mechanism of VPA on the recovery of neural function in TBI rats, provide a theoretical basis for clinical application.
objective
By establishing a rat model of closed craniocerebral injury, the effects of apoptosis, the expression of HSP70 and Caspase-3, TNF- - A and IL-1 beta in the brain tissue of rats at different time points, the dynamic changes of TNF- - A and IL-1 beta in brain tissue were observed, and the anti apoptosis of the rats with experimental craniocerebral injury was investigated. The protective effect and alleviating the effect of inflammatory reaction.
Method
The healthy SD male rats were randomly divided into normal control group, simple injury group and VPA treatment group. The rat model of closed craniocerebral injury was established, and the neurological disease severity score (Neurological Severity Scores, NSS) was used to evaluate the neural function of the rats at different time points in each group; the optical microscope was used to observe the morphology of the brain tissue. Change, in situ TDT labeling technique (TUNEL) was used to detect the cell apoptosis in the brain tissue of rats. The HSP70 and caspase-3 protein in the brain cells were detected by immunohistochemistry (SP method) and the TNF- alpha and IL- in the brain cells of each group were detected by immunofluorescence technique. The expression of 1 beta protein was detected by enzyme linked immunosorbent assay (ELISA), and the dynamic changes of TNF- alpha and IL-1 beta in the brain tissues were measured at different time points, and the statistical analysis was made.
Result
1. behavior test showed that the rats' behavioral score results were recovered in different degrees after VPA treatment, and there was no significant difference between the 3 groups after the trauma (P0.05). After VPA treatment, 7d, 14d, 21d, 28d, and VPA treatment group were better than the single pure injury group, and there was significant difference between the two groups (P0.05).
2. in situ end labeling (TUNEL) detection showed that the number of apoptotic cells in the cortex at all time points in the injured area of the injured group and the VPA group were higher than that in the normal control group, the difference was statistically significant (P0.05), the number of apoptotic cells at each time point in the.VPA treatment group was lower than that in the simple injury group, the difference was statistically significant (P0.05).
3. the immunohistochemical test showed that the mean gray value of HSP70 in the VPA treatment group was significantly lower than that of the simple injury group. The difference was statistically significant (P0.05) the mean gray value of Caspase-3 in the same time point of the.VPA treatment group was significantly higher than that of the simple injury group, and the difference had the significance of the unified planning (P0.05).
4. immunofluorescence assay: the number of TNF- alpha and IL-1 beta positive cells in the brain tissue at the same time point in the VPA treatment group was significantly lower than that in the simple injury group, and the difference was statistically significant (P0.05).
5. enzyme linked immunosorbent assay (ELISA) test: the concentration of TNF- alpha and IL-1 beta protein in the brain tissue at the same time point in the same time point after the model building and the VPA treatment group were higher than the normal control group, the difference was statistically significant (P0.05). The concentration of TNF- a and IL-1 beta protein in the brain tissue at the same time point in the VPA treatment group was lower than that of the simple injury group. Statistical significance (P0.05).
conclusion
VPA can improve the neurological function of rats with traumatic brain injury. VPA can resist apoptosis, protect nerve cells and reduce inflammatory reaction.

【學(xué)位授予單位】：新鄉(xiāng)醫(yī)學(xué)院
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2013
【分類(lèi)號(hào)】：R651.15

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