本文選題：顱腦創(chuàng)傷 + 糖代謝��； 參考：《重慶醫(yī)科大學(xué)》2014年博士論文

【摘要】：第一部分 大鼠腦損傷后急性期腦葡萄糖代謝和缺氧誘導(dǎo)因子、葡萄糖轉(zhuǎn)運(yùn)蛋白表達(dá)變化時(shí)序規(guī)律的研究 目的：通過(guò)檢測(cè)大鼠顱腦損傷模型急性期血糖、腦損傷區(qū)局部細(xì)胞外液(Extracellular Fluid,ECF)糖代謝指標(biāo)的變化以及損傷區(qū)局部皮層腦組織缺氧誘導(dǎo)因子-1α(Hypoxia-inducible Factor-1, HIF-1α)、葡萄糖轉(zhuǎn)運(yùn)蛋白1(glucose transporter-1,GLUT-1)和葡萄糖轉(zhuǎn)運(yùn)蛋白3(glucosetransporter3,GLUT-3)表達(dá)的變化，分析這些變化的時(shí)序性規(guī)律，以探討顱腦損傷后血糖控制的最佳時(shí)機(jī)。 方法：成年雄性SD大鼠35只，隨機(jī)分成7組，每組包含正常對(duì)照、顱腦創(chuàng)傷(traumatic brain injury,TBI)后不同時(shí)間點(diǎn)六組，每組5只。在傷后15m、30m、45m、75m、90m、3h、6h、12h、24h、48h、72h不同時(shí)間點(diǎn)測(cè)定血糖值并收集腦損傷區(qū)局部腦細(xì)胞間(ECF)透析液，監(jiān)測(cè)透析液葡萄糖含量([Glu]d)和乳酸含量([Lac]d)的變化，并在傷后3h、6h、12h、24h、48h、72h六個(gè)時(shí)間點(diǎn)斷頭取腦，采用RT-PCR和Western-blot法檢測(cè)局部皮層腦組織HIF-1α、GLUT-1、GLUT-3三者mRNA和蛋白的表達(dá)。 結(jié)果：1.顱腦創(chuàng)傷組傷后血糖水平逐漸上升，6h上升明顯，24h達(dá)到峰值，48h開(kāi)始下降，72h仍高于正常水平，對(duì)照組血糖水平變化不明顯。 2.顱腦創(chuàng)傷組腦細(xì)胞間透析液葡萄糖水平明顯降低，15～30min時(shí)間段達(dá)低谷，明顯低于對(duì)照組(P0.05)，其后逐漸回升，至75～90min時(shí)間段接近對(duì)照組；對(duì)照組腦細(xì)胞間透析液葡萄糖水平變化不明顯。 3.顱腦創(chuàng)傷組腦細(xì)胞間透析液乳酸水平明顯升高，傷后15～30min時(shí)間段達(dá)最高峰，可達(dá)基礎(chǔ)水平的2~3倍，明顯高于對(duì)照組(P0.05)，其后逐漸降低，至75~90min時(shí)間段接近對(duì)照組，而對(duì)照組腦細(xì)胞間透析液乳酸水平變化不明顯。 4.顱腦創(chuàng)傷組腦細(xì)胞間液葡萄糖（[Glu]d）、乳酸（[Lac]d）變化和對(duì)照組相比，變化具有統(tǒng)計(jì)學(xué)意義，P0.05。[Glu]d下降和[Lac]d上升呈相反趨勢(shì)變化。 5.顱腦創(chuàng)傷組腦損傷區(qū)皮層腦組織傷后3h可見(jiàn)有HIF-1α陽(yáng)性表達(dá)，后逐漸增強(qiáng)，48h后達(dá)高峰，和對(duì)照組比較具有顯著差異（P0.05），隨后表達(dá)逐漸減少；創(chuàng)傷組腦損傷區(qū)皮層腦組織GLUT-1表達(dá)傷后12h開(kāi)始降低，持續(xù)低至傷后72小時(shí)，和對(duì)照組比較具有顯著差異（P＜0.05）；創(chuàng)傷組腦損傷區(qū)皮層腦組織GLUT-3表達(dá)傷后12h開(kāi)始有增加，48h表達(dá)最明顯，72h開(kāi)始下降，但仍高于正常水平，和對(duì)照組比較有顯著差異（P＜0.05）。對(duì)照組對(duì)應(yīng)區(qū)皮層腦組織GLUT-1、GLUT-3表達(dá)在各時(shí)間段變化不明顯，偶見(jiàn)HIF-1α陽(yáng)性表達(dá)。 結(jié)論：顱腦創(chuàng)傷后血糖、腦損傷區(qū)皮層腦細(xì)胞間液糖、乳酸及HIF-1α、GLUT-1、GLUT-3的變化在時(shí)間上具有一定特點(diǎn)：損傷區(qū)皮層腦細(xì)胞糖代謝最先發(fā)生變化，隨后HIF-1α開(kāi)始表達(dá)，血糖升高，最后，GLUT-1、GLUT-3逐漸出現(xiàn)表達(dá)變化。這種變化可能是機(jī)體對(duì)局部缺氧代償和葡萄糖轉(zhuǎn)運(yùn)、利用障礙的結(jié)果，作用機(jī)制復(fù)雜。傷后6～12小時(shí)是腦創(chuàng)傷后局部腦細(xì)胞糖代謝相關(guān)基因、蛋白發(fā)生變化的關(guān)鍵時(shí)期，，可能為控制性降糖預(yù)防繼發(fā)性腦損傷發(fā)生的最佳時(shí)間段。 第二部分 高靈敏葡萄糖生物傳感器的制備及其在顱腦損傷患者腦脊液糖含量檢測(cè)中的運(yùn)用 目的：制備基于納米材料的高靈敏葡萄糖生物傳感器，探討高靈敏葡萄糖生物傳感器在腦脊液葡萄糖水平監(jiān)測(cè)中運(yùn)用的可行性。 方法：采用高靈敏葡萄糖生物傳感器對(duì)5例顱腦損傷患者所采集的腦脊液中糖含量進(jìn)行檢測(cè)，同時(shí)運(yùn)用實(shí)驗(yàn)室葡萄糖氧化酶法檢測(cè)，對(duì)兩種方法的檢測(cè)范圍、準(zhǔn)確性、靈敏度、精密度以及抗干擾能力進(jìn)行比較。 結(jié)果： 1.高靈敏葡萄糖生物傳感器響應(yīng)電流值隨著葡萄糖從1.2×10-6到1.6×10-3M的加入而呈線性增加，相關(guān)系數(shù)為0.998（n=20）。檢測(cè)限為4.0×10-7M,信噪比為3，高靈敏葡萄糖生物傳感器檢測(cè)范圍廣，靈敏度高。 2.組內(nèi)和組間精密度的相對(duì)標(biāo)準(zhǔn)偏差分別為3.8%和5.1%,高靈敏葡萄糖生物傳感器檢測(cè)具有良好的準(zhǔn)確性和重現(xiàn)性。 3.在含0.2mM葡萄糖的PBS中加入一定濃度干擾物質(zhì)（抗壞血酸，尿酸，對(duì)乙酰氨基酚以及L-半胱氨酸），對(duì)葡萄糖的檢測(cè)無(wú)干擾，高靈敏葡萄糖生物傳感器具有良好的抗干擾能力。 4.檢測(cè)結(jié)果的準(zhǔn)確性與臨床檢驗(yàn)方法（GLU-GOD-PAP方法）進(jìn)行對(duì)比，樣本稀釋10倍，所得結(jié)果的相對(duì)偏差為-6.45%到8.69%之間，高靈敏葡萄糖生物傳感器檢測(cè)精密度高。 結(jié)論： 運(yùn)用高靈敏葡萄糖生物傳感器對(duì)顱腦損傷患者腦脊液葡萄糖水平檢測(cè)響應(yīng)快，靈敏度、精密度高，檢測(cè)結(jié)果準(zhǔn)確，抗干擾能力強(qiáng)，和傳統(tǒng)葡萄糖氧化酶法檢測(cè)相比更靈敏、快捷�？蛇M(jìn)一步用于顱腦外傷后患者腦脊液葡萄糖水平的動(dòng)態(tài)監(jiān)測(cè)。 第三部分 顱腦損傷后動(dòng)態(tài)監(jiān)測(cè)顱內(nèi)壓、灌注壓及血腦屏障指數(shù)的臨床意義 目的：通過(guò)檢測(cè)、計(jì)算顱腦外傷后血腦屏障指數(shù)，并動(dòng)態(tài)監(jiān)測(cè)其變化，探討血腦屏障指數(shù)與顱腦損傷嚴(yán)重程度及病情演變之間是否存在相關(guān)性。 方法：對(duì)30例中、重型顱腦損傷患者的血腦屏障指數(shù)進(jìn)行動(dòng)態(tài)監(jiān)測(cè)結(jié)合GCS(glasgow coma scale，GCS)評(píng)分、顱內(nèi)壓及灌注壓，分析血腦屏障指數(shù)和顱腦損傷嚴(yán)重程度和病情演變之間的相關(guān)性。 結(jié)果：所有患者血腦屏障指數(shù)均高于文獻(xiàn)中的正常范圍。血腦屏障指數(shù)和顱腦損傷的嚴(yán)重程度（GCS評(píng)分）呈正相關(guān)，和高顱壓及低灌注壓累及時(shí)間（pressure times Time dose,PTD）呈正相關(guān)，和病情演變具有一致性。 結(jié)論：顱腦損傷后血腦屏障通透性發(fā)生改變，影響其功能。血腦屏障指數(shù)的動(dòng)態(tài)監(jiān)測(cè)結(jié)合顱內(nèi)壓、灌注壓可以客觀反映病情的演變，血腦屏障指數(shù)可以作為臨床監(jiān)測(cè)指標(biāo)之一。 第四部分 顱內(nèi)壓及腦脊液生物標(biāo)記物S100β和NSE聯(lián)合監(jiān)測(cè)在顱腦損傷中的臨床意義 目的：通過(guò)動(dòng)態(tài)監(jiān)測(cè)顱腦損傷后腦脊液S100β(S100β protein)和NSE(neuron-specific enolase,NSE)水平，分析二者和高顱壓及腦低灌注壓的相關(guān)性，探討監(jiān)測(cè)腦脊液S100β和NSE水平在評(píng)估顱腦損傷的嚴(yán)重程度及病情進(jìn)展中的意義。 方法：對(duì)重慶醫(yī)科大學(xué)附屬第二醫(yī)院和重慶市江津區(qū)中心醫(yī)院2013年1月至2013年10月符合入選標(biāo)準(zhǔn)的顱腦損傷患者進(jìn)行前瞻性的研究。所有符合入選標(biāo)準(zhǔn)的病例從腦室引流管收集腦脊液，檢測(cè)S100β和NSE水平，每天2次，取平均值，一共7天。記錄每小時(shí)ICP、CCP，取超過(guò)臨界值壓力的均值，將壓力均值和24小時(shí)內(nèi)ICP20mmHg，CPP60mmHg累計(jì)時(shí)間相乘，看作高顱壓、低灌注的累及時(shí)間（pressure times Time dose,PTD）PTD20、PTD60。比較腦脊液S100β和NSE水平和PTD20、PTD60。分析腦脊液S100β、NSE的水平和ICP及CCP的動(dòng)態(tài)變化的關(guān)系。 結(jié)果：36例病人被納入。中型組(GCS9～12)20例，重型組(GCS4～8)16例。采集腦脊液標(biāo)本，對(duì)這些標(biāo)本進(jìn)行了檢測(cè)，結(jié)果所有患者S100β和NSE水平均增高，和顱腦損傷嚴(yán)重程度（GCS評(píng)分）呈正相關(guān)。同時(shí)S100β水平增高和顱內(nèi)壓20mmHg、灌注壓60mmHg累及時(shí)間關(guān)系密切（PTD20:r=0.35,P 0.001;PTD60:r=0.24,P 0.001）；NSE水平增高和顱內(nèi)壓20mmHg累及時(shí)間PTD20呈弱相關(guān)(r=0.17, P=0.01)，和灌注壓60mmHg累及時(shí)間PTD60關(guān)系密切(r=0.20, P0.01)。腦脊液S100β和NSE水平的變化和顱腦損傷嚴(yán)重程度及病情演變具有一致性。 結(jié)論：顱腦損傷后腦脊液S100β和NSE水平的增高程度反映了顱腦損傷嚴(yán)重程度，腦脊液S100β和NSE水平的動(dòng)態(tài)變化反映了病情的演變。對(duì)二者腦脊液水平的變化結(jié)合顱內(nèi)壓、灌注壓進(jìn)行動(dòng)態(tài)監(jiān)測(cè)可以客觀反映病情的進(jìn)展，可以在出現(xiàn)明顯的繼發(fā)性腦損傷臨床癥狀之前預(yù)測(cè)繼發(fā)性腦損傷的發(fā)生。對(duì)中、重型顱腦損傷患者實(shí)行S100β、NSE聯(lián)合ICP、CCP的動(dòng)態(tài)監(jiān)測(cè)具有重要臨床意義。
[Abstract]:Part one
Timing of brain glucose metabolism and expression of hypoxia inducible factor and glucose transporter in rats with acute brain injury
Objective: to detect the changes of glucose metabolism in the local extracellular fluid (Extracellular Fluid, ECF) and the hypoxia inducible factor -1 alpha (Hypoxia-inducible Factor-1, HIF-1 alpha), glucose transporter 1 (glucose transporter-1, GLUT-1) and grape (glucose transporter-1, GLUT-1) in the local cortical tissue of the injured area by detecting the acute blood glucose in the rat brain injury model. The changes of glucosetransporter3 (GLUT-3) expression were analyzed, and the timing rules of these changes were analyzed to explore the best timing of blood glucose control after craniocerebral injury. 3.
Methods: 35 adult male SD rats were randomly divided into 7 groups, each group contained normal control, traumatic brain injury (TBI), six groups at different time points, 5 in each group. After the injury, 15m, 30m, 45m, 75m, 90m, 3h, 6h, etc. were determined at different time points and collected local brain cells in the brain injury area and monitored dialysate. The changes of glucose content ([Glu]d) and lactic acid content ([Lac]d) were observed at six time points of 3H, 6h, 12h, 24h, 48h and 72h after injury, and HIF-1 alpha, GLUT-1, and protein expression were detected by RT-PCR and Western-blot methods.
Results: 1. after craniocerebral trauma group, blood glucose level increased gradually, 6h increased obviously, 24h reached peak value, 48h began to decrease, 72h was still higher than normal level, and the blood sugar level in control group was not obvious.
2. the level of glucose in the dialysate between brain cells in the brain trauma group decreased significantly, and the time period of 15 ~ 30min was lower than that of the control group (P0.05), and then gradually recovered to the control group from 75 to 90min, and the glucose level of the dialysate between the cerebral cells of the control group was not obvious.
3. the level of lactic acid in the dialysate between brain cells in the traumatic brain injury group was significantly higher, the peak of the 15 ~ 30min period after injury reached the peak, up to 2~3 times of the basic level, obviously higher than that of the control group (P0.05), and then gradually decreased to the 75~90min time period to close to the control group, but the change of lactic acid level of the dialysate between the brain cells of the control group was not obvious.
4. the changes of glucose ([Glu]d) and lactic acid ([Lac]d) in the brain cells of the traumatic brain injury group were statistically significant compared with the control group. The decrease of P0.05.[Glu]d and the rise of [Lac]d were in the opposite direction.
5. the positive expression of HIF-1 alpha was found in 3h after brain injury in brain injury area, and then increased gradually, and then reached the peak after 48h. There was a significant difference between the control group and the control group (P0.05), and then the expression gradually decreased. The 12h began to decrease after GLUT-1 expression in the cortex brain tissue of the traumatic brain injury group and was lower to 72 hours after the injury, compared with the control group. There was significant difference (P < 0.05). After the GLUT-3 expression of the cortical brain tissue in the trauma group, the 12h began to increase, the expression of 48h was the most obvious, the 72h began to decline, but it was still higher than the normal level, and there was a significant difference between the control group and the control group (P < 0.05). The expression of GLUT-1 in the corresponding cortex and brain group of the control group was not obvious in each time period, occasionally HI. The expression of F-1 alpha was positive.
Conclusion: after craniocerebral trauma, the changes of sugar, lactate and HIF-1 alpha, GLUT-1, GLUT-3 in cerebral cortex and cerebral cells in the brain injury area have some characteristics in time: the glucose metabolism in the cortex brain cells of the injured area changes first, then the HIF-1 alpha begins to express, the blood sugar rises, and finally, GLUT-1, GLUT-3 changes. This change may be The mechanism of the body to local anoxic compensation and glucose transport and the use of obstacles is complicated. 6~12 hours after injury is the key period of the related genes of glucose metabolism in local brain cells after traumatic brain injury and the best time for controlling the occurrence of secondary brain injury by controlling hypoglycemic control.
The second part
Preparation of high sensitive glucose biosensor and its application in detecting CSF sugar content in patients with craniocerebral injury
Objective: to prepare a highly sensitive glucose biosensor based on nanomaterials, and to explore the feasibility of using highly sensitive glucose biosensor to monitor the glucose level in cerebrospinal fluid.
Methods: the glucose content in cerebrospinal fluid of 5 patients with craniocerebral injury was detected by high sensitive glucose biosensor, and the glucose oxidase method was used in the laboratory. The detection range, accuracy, sensitivity, precision and anti-interference ability of the two methods were compared.
Result錛

本文編號(hào)：1821943