本文選題：TBI + TAK1　； 參考：《南京大學》2013年碩士論文

【摘要】：第一部分 題目：創(chuàng)傷性腦損傷后TAKl的表達時相和定位 目的：研究TAK1在TBI后動態(tài)表達和分布情況,探討TAK1通路和TBI的相互關系,以期為TBI的治療提供一新的治療靶點。 方法：1、實驗模型建立和分組：選擇健康成年SD雄性大鼠60只,體重為250-300g,采用改良的Feeney法于右側開顱制作TBI模型。將動物按損傷不同時間點分為6組,分別為假手術組、傷后1h組、傷后3h組、傷后6h組、傷后12h組、傷后24h組,各組動物均為10只。 2、TBI后TAK1及p-TAK1蛋白表達測定：按不同時間點分別提取大鼠挫傷灶周圍皮層腦組織,采用western blot方法檢測TAK1和p-TAK1蛋白的動態(tài)表達。 3、采用免疫組化法觀察：TAK1在傷灶周圍腦組織的表達和分布情況。 4、采用免疫熒光技術觀察：TAK1在TBI后損傷灶周圍皮層中不同細胞表達和分布情況。 結果：1、TBI組術后各時間點TAK1和p-TAK1的蛋白水平24h內(nèi)持續(xù)升高,24h明顯升高(p0.05)； 2、免疫組化結果顯示：假手術組中,TAK1在正常腦組織中有表達,主要分布于核周,外傷后胞漿表達量明顯升高；熒光顯微鏡結果顯示,在正常腦組織中,TAK1的表達主要位于神經(jīng)元中,在星形膠質(zhì)細胞中微量表達,損傷后TAK1在傷灶周圍皮層的神經(jīng)元和膠質(zhì)細胞的胞漿中的表達量明顯升高。 結論：1、TAK1在TBI早期即可被激活。 2、TBI后TAK1表達主要位于神經(jīng)元和膠質(zhì)細胞胞漿中,且其激活的區(qū)域與腦外傷后損傷區(qū)域相一致。 第二部分 題目：OZ對TBI的神經(jīng)保護作用及機制研究 目的：通過研究OZ對TBI后TAK1通路的調(diào)控,探討OZ對TBI的神經(jīng)保護作用及其具體分子調(diào)控機制,以期為臨床治療TBI提供新的策略,同時也為OZ臨床應用提供理論基礎和實驗依據(jù) 方法：1、實驗動物模型和分組：采用健康SD大鼠,運用改良的Feeney法于右側開顱制作TBI模型。實驗動物分組：大鼠分為五組：假手術組、外傷組、外傷+溶劑組、外傷+藥物組(2μg,10μg和20μg)。溶劑和藥物在損傷后10mim或3h通過側腦室注射5μ1劑量。 2、神經(jīng)功能評分：采用平衡木實驗評價實驗動物TBI后7天內(nèi)神經(jīng)功能缺損狀況。 3、神經(jīng)元凋亡檢測：取各組實驗動物大腦組織,石蠟固定,切片,TUNEL法檢測神經(jīng)元凋亡情況。 4、炎性因子檢測：取各組傷灶周圍腦組織,采用ELISA法檢測炎性因子的表達量； 5、TAK1活性及通路下游NF-κB和AP-1的活性：取各組實驗動物傷灶周圍皮層組織,采用western blot方法檢測TAK1和p-TAK1的蛋白含量,EMSA方法檢測NF-κB和AP。1的活性。 6、尼氏染色法檢測：外傷后7天神經(jīng)元的存活情況。 結果：1、與假手術組相比,TBI后大鼠表現(xiàn)出明顯的神經(jīng)功能缺損、神經(jīng)元凋亡,兩組間有顯著性差異(P0.05)；而OZ治療能明顯改善大鼠神經(jīng)功能評分、減輕神經(jīng)元的凋亡,兩組間有顯著性差異(P0.05)。 2、TBI后損傷灶周圍腦組織中TAK1蛋白表達增強；0Z明顯下調(diào)TAK1蛋白表達。 3、與TBI組相比,OZ明顯降低了NF-kB和AP-1的活性。 4、假手術組大鼠腦組織中TNF-α和IL-1p蛋白水平較低；腦損傷組中TNF-α和幾.1p蛋白水平明顯上調(diào),與TBI組相比,TNF-α和IL.1p蛋白水平明顯下降。 5、與假手術組相比,TBI后7天尼氏染色損傷神經(jīng)元個數(shù)明顯增加,而OZ治療能明顯改善神經(jīng)元的損傷。 結論：1、OZ能減輕TBI后繼發(fā)性損傷,對TBI有神經(jīng)保護作用。 2、OZ神經(jīng)保護作用是通過抑制NF-κB和AP.1通路,抑制炎性因子的產(chǎn)生從而減輕炎性損傷來實現(xiàn)。 3、以TAK1為靶點的藥物治療對臨床TBI的防治具有良好的應用前景。 第三部分 題目：TAKl在TBI患者挫傷區(qū)皮層的表達和定位 目的：研究TAK1在TBI患者挫傷區(qū)皮層的表達及分布。 方法挫傷區(qū)皮層標本來自5例TBI患者,傷后取標本時間24h,陰性對照組為3例腫瘤患者的正常腦組織(由于手術入路要求切除的),通過免疫組織化學和免疫熒光方法檢測TAK1在挫傷皮層的表達和定位。 結果：與陰性對照組相比,免疫組化結果顯示挫傷區(qū)皮層TAK1的表達量明顯升高；免疫熒光結果顯示TAK1在TBI患者挫傷區(qū)皮層的神經(jīng)元和膠質(zhì)細胞中均有表達。 結論：TAK1在挫傷區(qū)皮層的表達量升高,提示針對TAK1的靶點治療在TBI的臨床治療中具有重要的臨床應用價值。
[Abstract]:Part one
Title: expression and location of TAKl after traumatic brain injury
Objective: To study the dynamic expression and distribution of TAK1 after TBI, and to explore the relationship between TAK1 pathway and TBI, in order to provide a new therapeutic target for TBI treatment.
Methods: 1, the establishment and grouping of the experimental model: 60 healthy adult SD male rats were selected and the weight was 250-300g. The modified Feeney method was used to make the TBI model on the right craniotomy. The animals were divided into 6 groups according to the different time points of injury. They were sham operation group, 1H group after injury, 3h group after injury, 6h group after injury, 12h group after injury, group 24h after injury, all the animals of each group were 1. 0.
2, after TBI, the expression of TAK1 and p-TAK1 protein was measured. The brain tissue around the rat contusion was extracted at different time points, and the dynamic expression of TAK1 and p-TAK1 protein was detected by Western blot.
3, immunohistochemical staining was used to observe the expression and distribution of TAK1 in the brain tissue around the lesion.
4, immunofluorescence technique was used to observe the expression and distribution of TAK1 cells in the cortex around the lesion after TBI.
Results: 1. The protein levels of TAK1 and p-TAK1 in group TBI increased continuously during 24h, and 24h increased significantly (P0.05).
2, the results of immunohistochemistry showed that in the sham operation group, TAK1 was expressed in normal brain tissue, mainly distributed in the perinuclear tissue, and the expression of cytoplasm was significantly increased after trauma. The results of fluorescence microscopy showed that in normal brain tissue, the expression of TAK1 was mainly in the neurons, and in the astrocytes, the TAK1 was in the surrounding skin after injury. The expression levels of neurons and glial cells in the cytoplasm increased significantly.
Conclusion: 1, TAK1 can be activated in the early stage of TBI.
2, the expression of TAK1 was mainly located in cytoplasm of neurons and glial cells after TBI, and the region of activation was consistent with that of post traumatic brain injury.
The second part
Title: the neuroprotective effect and mechanism of OZ on TBI
Objective: To explore the neuroprotective effect of OZ on TBI and its specific molecular regulation mechanism by studying the regulation of OZ on the TAK1 pathway after TBI, so as to provide a new strategy for the clinical treatment of TBI, and to provide theoretical basis and experimental basis for the clinical application of OZ.
Methods: 1, experimental animal models and groups: using healthy SD rats, using the modified Feeney method to make the TBI model on the right craniotomy. Experimental animals were divided into five groups: sham operation group, trauma group, trauma + solvent group, trauma + drug group (2 mu g, 10 mu g and 20 micron g). The solution and drug were injected 5 mu through the lateral ventricle of 10mim or 3h after injury. Dose.
2, neurological function score: the balance function test was used to evaluate the neurological deficit in 7 days after TBI.
3, neuron apoptosis detection: take brain tissues of experimental animals, paraffin fixed and slice, and detect apoptosis of neurons by TUNEL.
4, inflammatory factors were detected: the brain tissue around each lesion was collected and the expression of inflammatory factors was detected by ELISA.
5, the activity of TAK1 and the activity of NF- kappa B and AP-1 downstream of the pathway: the protein content of TAK1 and p-TAK1 was detected by Western blot method, and the activity of NF- kappa B and AP.1 were detected by Western blot method.
6, Nissl staining was used to detect the survival of neurons on the 7 day after trauma.
Results: 1, compared with the sham operation group, the rats showed obvious nerve function defect and neuron apoptosis after TBI, and there was significant difference between the two groups (P0.05), and OZ treatment could obviously improve the neural function score of rats and reduce the apoptosis of the neurons. There was significant difference between the two groups (P0.05).
2, after TBI, the expression of TAK1 protein was increased in the brain tissue around the lesion. 0Z significantly decreased the expression of TAK1 protein.
3, compared with group TBI, OZ significantly reduced the activity of NF-kB and AP-1.
4, the level of TNF- alpha and IL-1p protein in the rat brain tissue of the sham operation group was low, and the level of TNF- alpha and several.1p protein in the brain injury group was obviously up, and the level of TNF- alpha and IL.1p protein decreased significantly compared with the TBI group.
5, compared with the sham operation group, the number of Nissl stained neurons increased significantly on the 7 day after TBI, while OZ treatment significantly improved neuronal damage.
Conclusion: 1, OZ can alleviate secondary injury after TBI and has neuroprotective effect on TBI.
2, the neuroprotective effect of OZ is achieved by inhibiting the NF- kappa B and AP.1 pathway, inhibiting the production of inflammatory factors and reducing inflammatory injury.
3, TAK1 targeted drug therapy has a good application prospect in the prevention and treatment of clinical TBI.
The third part
Title: expression and localization of TAKl in the contusion cortex of TBI patients
Objective: To study the expression and distribution of TAK1 in the contusion area of TBI patients.
Methods the cortical specimens from the contusion area were from 5 patients with TBI. The time was 24h after injury, and the normal brain tissue of the negative control group was 3 cases of the tumor patients (due to the surgical procedure). The expression and location of TAK1 in the contusion cortex were detected by immunohistochemistry and immunofluorescence.
Results: compared with the negative control group, the immunohistochemical results showed that the expression of TAK1 in the contusion area was significantly increased, and the results of immunofluorescence showed that TAK1 was expressed in the neurons and glia cells in the contusion area of TBI patients.
Conclusion: the expression level of TAK1 in the cortex of the contusion area is higher, suggesting that targeted therapy for TAK1 has important clinical value in the clinical treatment of TBI.
【學位授予單位】：南京大學
【學位級別】：碩士
【學位授予年份】：2013
【分類號】：R651.15

【共引文獻】

相關期刊論文 前10條

1 陳大帥;周思遠;唐宏智;李瑛;;針刺調(diào)節(jié)核轉錄因子-κB信號通路的實驗研究進展[J];中華中醫(yī)藥雜志;2013年10期

2 肖愛嬌;羅小泉;;艾灸對腦缺血再灌注損傷模型大鼠大腦皮質(zhì)NF-κB p65蛋白表達的影響[J];中華中醫(yī)藥雜志;2013年12期

3 王靖坤;王順妮;陳紅巖;范薇薇;閔太善;盧大儒;陳浩明;;慢病毒介導的Jab1基因表達干擾在乳腺癌細胞中功能的初步研究[J];復旦學報(自然科學版);2013年06期

4 朱海濤;林江凱;;顱腦創(chuàng)傷后神經(jīng)炎癥反應中的Toll樣受體[J];創(chuàng)傷外科雜志;2014年01期

5 方梅榮;金英順;金吉哲;崔鎮(zhèn)花;金海峰;鄭海蘭;李錦姬;姜玉姬;金華;李燦;;骨橋蛋白和核轉錄因子在慢性環(huán)孢素A腎毒性中的作用[J];第二軍醫(yī)大學學報;2014年05期

6 馬永達;王旭輝;陳強;;糖皮質(zhì)激素調(diào)節(jié)創(chuàng)傷后神經(jīng)元胞膜鈣泵研究現(xiàn)狀[J];創(chuàng)傷與急危重病醫(yī)學;2015年01期

7 張文艷;孫寶飛;余資江;余彥;肖朝倫;羅時鵬;令狐艷;楊丹;;原花青素對小鼠腦缺血再灌注損傷的神經(jīng)保護機制[J];貴陽醫(yī)學院學報;2015年04期

8 趙衛(wèi)海;海陳煉;余國棟;;血清TNF-αIL-1IL-6IL-8在急性顱腦損傷后含量變化及意義[J];中國實用神經(jīng)疾病雜志;2013年16期

9 黃強;戴偉民;揭園慶;聶俊;王小芳;胡永亮;余小明;宋光太;金濤;吳來德;;衢州地區(qū)急性顱腦創(chuàng)傷患者院前及急診救治現(xiàn)狀分析[J];臨床神經(jīng)外科雜志;2013年04期

10 于建云;;多重性腦震蕩及其后遺癥研究進展[J];昆明醫(yī)科大學學報;2013年10期

相關博士學位論文 前10條

1 高麗;血管緊張素-（1-7）在高血壓及腦缺血損傷中的作用及機制研究[D];南京醫(yī)科大學;2013年

2 陳寅螢;多維度多組分干預腦缺血藥理機制的比較研究[D];中國中醫(yī)科學院;2013年

3 馮秀麗;法氏囊活性肽的分離鑒定及其免疫調(diào)節(jié)作用機制研究[D];南京農(nóng)業(yè)大學;2012年

4 楊子彥;三角帆蚌肽聚糖識別蛋白和谷胱甘肽硫轉移酶的基因克隆和功能分析[D];華中科技大學;2013年

5 伍列林;Let-7g下調(diào)HMGA2抑制原發(fā)性肝細胞癌增殖、侵襲的作用及機制研究[D];中南大學;2013年

6 趙永博;SIRT1在創(chuàng)傷性腦損傷后的功能及機制研究[D];第四軍醫(yī)大學;2013年

7 楊尚武;Nek9和ERK8在小鼠卵母細胞成熟中的作用和在早期胚胎中定位[D];南方醫(yī)科大學;2013年

8 秦文熠;電針及NBD多肽抑制局灶腦缺血/再灌注大鼠大腦皮質(zhì)及海馬CA1區(qū)炎癥反應及其機制的研究[D];重慶醫(yī)科大學;2013年

9 張峗;TLR2在骨髓間充質(zhì)干細胞修復缺氧缺血性腦損傷中的作用及機制[D];重慶醫(yī)科大學;2013年

10 周帥;載脂蛋白E及其基因多態(tài)性對星形膠質(zhì)細胞缺氧性損傷的影響及相關機制研究[D];重慶醫(yī)科大學;2013年

相關碩士學位論文 前10條

1 凌海平;中性粒細胞彈性蛋白酶在小鼠創(chuàng)傷性腦損傷后炎癥反應中作用機制的研究[D];南方醫(yī)科大學;2012年

2 錢偉;NF-κB在大鼠顱腦損傷后繼發(fā)性腦損傷中的作用及意義[D];蘇州大學;2013年

3 陳永蘭;p38MAPK信號通路對小膠質(zhì)細胞功能及NGF、TNFα分泌的影響[D];大連醫(yī)科大學;2012年

4 王澤昒;西維因及主要降解產(chǎn)物1-萘酚對雄性羅非魚雌激素效應的研究[D];南京農(nóng)業(yè)大學;2011年

5 楊岳;中國荷斯坦牛C4A基因的遺傳變異及其作為乳腺炎抗性標記的可行性分析[D];南京農(nóng)業(yè)大學;2011年

6 馬巖巖;三七皂甙成分對脂多糖誘導巨噬細胞炎性因子的影響及其機制的研究[D];暨南大學;2013年

7 羅流濤;骨髓間充質(zhì)干細胞對阿霉素大鼠足細胞骨架F-actin的影響及機制探討研究[D];福建醫(yī)科大學;2013年

8 蔡理;腦損傷后JNK3基因參與神經(jīng)細胞凋亡及神經(jīng)功能恢復[D];昆明醫(yī)科大學;2013年

9 趙明明;機械性神經(jīng)元損傷后自噬的功能及Homer1a通過PI3K/Akt通路對其調(diào)節(jié)作用[D];第四軍醫(yī)大學;2013年

10 明月;綠僵菌絲氨酸—蘇氨酸蛋白激酶基因MaSnf1和MaHog1的克隆及功能分析[D];重慶大學;2013年

，

本文編號：1964663