本文選題：神經(jīng)退行性病變 + 小膠質(zhì)細(xì)胞��； 參考：《第四軍醫(yī)大學(xué)》2012年碩士論文

【摘要】：【研究背景】 神經(jīng)退行性病變是一類(lèi)年齡相關(guān)的以神經(jīng)元功能和結(jié)構(gòu)喪失甚至死亡為病理特征的疾病，典型疾病如阿茲海默癥（Alzheimer’s disease,AD），帕金森癥（Parkinson’s disease, PD）等，臨床癥狀根據(jù)損傷的不同類(lèi)型表現(xiàn)出不同的行為、精神、認(rèn)知、運(yùn)動(dòng)性障礙。在病理學(xué)上，炎癥與氧化應(yīng)激對(duì)神經(jīng)退行性病變有十分重要的作用。 小膠質(zhì)細(xì)胞作為神經(jīng)系統(tǒng)的固有免疫單位，和神經(jīng)退行性病變有非常重要的聯(lián)系。靜息的小膠質(zhì)細(xì)胞對(duì)于神經(jīng)系統(tǒng)有支持、監(jiān)視、修復(fù)等功能；小膠質(zhì)細(xì)胞對(duì)腦內(nèi)環(huán)境變化十分敏感，一些刺激能使小膠質(zhì)細(xì)胞活化，小膠質(zhì)細(xì)胞活化可以產(chǎn)生很多炎癥趨化因子和自由基，這些物質(zhì)大部分具有細(xì)胞毒性，蓄積過(guò)高可以使神經(jīng)元退化、變性、死亡，進(jìn)一步會(huì)導(dǎo)致神經(jīng)退行性病變。多種信號(hào)通路都參與了小膠質(zhì)細(xì)胞的活化過(guò)程，如JAK-STAT、MAPK等，且不同的通路之間互相聯(lián)系，互相影響，信號(hào)通路調(diào)控對(duì)于調(diào)節(jié)小膠質(zhì)細(xì)胞狀態(tài)具有重要意義。因此找到抑制小膠質(zhì)細(xì)胞的活化的方法和途徑在預(yù)防和治療神經(jīng)退行性病變中有重要的意義。以小膠質(zhì)細(xì)胞作為神經(jīng)退行性病變的治療靶點(diǎn)已經(jīng)成為一種可行的方式，，目前已證實(shí)多種藥物均能調(diào)控小膠質(zhì)細(xì)胞的活化，且具有一定的臨床治療或預(yù)防作用。烏司他�。║linastain，UTI）是一種蛋白酶抑制劑，在腦及其他多種器官系統(tǒng)中均被證實(shí)具有明確的抗炎抗氧化作用，它對(duì)于小膠質(zhì)細(xì)胞活化的過(guò)程是否有調(diào)控作用還未被證實(shí)，本文將基于此問(wèn)題展開(kāi)一些討論。 【目的】 通過(guò)利用脂多糖（lipopolysaccharide，LPS）建立體內(nèi)外小膠質(zhì)細(xì)胞活化的模型，觀(guān)察UTI治療對(duì)于小膠質(zhì)細(xì)胞活化的作用和對(duì)學(xué)習(xí)記憶功能的影響。并進(jìn)一步探討其中的信號(hào)通路機(jī)制。 【方法】 1、利用LPS注射大鼠建立小膠質(zhì)細(xì)胞活化的體內(nèi)模型。 2、利用LPS刺激BV2細(xì)胞建立小膠質(zhì)細(xì)胞的體外模型。 3、 Morris水迷宮（Morris Water Maze，MWM）觀(guān)察大鼠學(xué)習(xí)記憶功能的變化。 4、免疫組織化學(xué)觀(guān)察小膠質(zhì)細(xì)胞形態(tài)學(xué)的變化。 5、 ELISA檢測(cè)TNF-α和IL-1β的釋放。 6、 Western Blot檢測(cè)相關(guān)信號(hào)通路的變化。 【結(jié)果】 1、 LPS誘導(dǎo)小膠質(zhì)細(xì)胞細(xì)胞活化模型的建立 體內(nèi)實(shí)驗(yàn)部分，LPS腹腔注射后，大鼠的Morris水迷宮成績(jī)下降，免疫組織化學(xué)可觀(guān)察到腦內(nèi)海馬區(qū)大量活化的小膠質(zhì)細(xì)胞，腦組織ELISA檢測(cè)提示TNF-α和IL-1β的含量增高。體外實(shí)驗(yàn)部分，LPS刺激可以使BV2細(xì)胞TNF-α和IL-1β的釋放增高。 2、 UTI對(duì)LPS誘導(dǎo)的小膠質(zhì)細(xì)胞活化的抑制效應(yīng) 體內(nèi)試驗(yàn)部分，100kU/kg的UTI對(duì)于LPS注射引起的Morris水迷宮成績(jī)下降有改善作用，免疫組織化學(xué)結(jié)果顯示小膠質(zhì)細(xì)胞的活化減少，ELISA結(jié)果提示TNF-α和IL-1β含量下降。體外實(shí)驗(yàn)部分，UTI處理的BV2細(xì)胞在LPS刺激下TNF-α和IL-1β的釋放減少。 3、 UTI抑制LPS誘導(dǎo)的小膠質(zhì)細(xì)胞活化的信號(hào)轉(zhuǎn)導(dǎo)機(jī)制 體內(nèi)外實(shí)驗(yàn)均發(fā)現(xiàn)，在LPS的刺激可誘導(dǎo)海馬組織和BV2細(xì)胞的MAPKs和JAK-STAT通路活性上調(diào)，在UTI處理后他們的活性下降。通過(guò)抑制部分MAPKs通路的表達(dá)可以抑制LPS誘導(dǎo)的STAT3活性的升高。 【結(jié)論】 1、 LPS可以使小膠質(zhì)細(xì)胞活化，炎癥因子釋放增多，并影響學(xué)習(xí)記憶功能。 2、 UTI對(duì)LPS引起的小膠質(zhì)細(xì)胞活化有抑制作用，從而降低炎癥因子的釋放，改善學(xué)習(xí)記憶功能。 3、 UTI對(duì)LPS引起的小膠質(zhì)細(xì)胞活化的抑制作用可能是通過(guò)抑制MAPKs和JAK-STAT通路實(shí)現(xiàn)的。在這個(gè)過(guò)程中，MAPKs和JAK-STAT通路存在聯(lián)系，MAPKs參與調(diào)控STAT3的磷酸化。 綜上所述，UTI對(duì)于神經(jīng)系統(tǒng)有一定的保護(hù)作用，可望作為一種新的藥物用于神經(jīng)退行性病變的治療和預(yù)防。
[Abstract]:[research background]
Neurodegenerative disease is a class of age-related diseases characterized by neuronal function and loss of structure or even death. Typical diseases such as Alzheimer 's disease (AD), Parkinson's (Parkinson' s disease, PD) and so on. Clinical symptoms show different behavior, mental, cognitive, and exercise according to the different types of damage. In pathology, inflammation and oxidative stress play an important role in neurodegenerative diseases.
Microglia, as an intrinsic immune unit of the nervous system, has a very important relationship with neurodegenerative diseases. Resting microglia can support, monitor, repair and other functions of the nervous system. Microglia are sensitive to changes in the brain environment. Some stimulations can activate microglia and microglia can be activated. Many inflammatory chemokines and free radicals are produced. Most of these substances are cytotoxic. Excessive accumulation of these substances can cause degeneration, degeneration, death, and further cause neurodegenerative diseases. Many signal pathways are involved in the activation process of microglia, such as JAK-STAT, MAPK, and the interconnections between different pathways. It is of great significance to regulate the state of microglia. Therefore, it is important to find ways and ways to prevent the activation of microglia in the prevention and treatment of neurodegenerative diseases. It is a feasible way to treat microglia as a target for treatment of neurodegenerative diseases. It has been confirmed that a variety of drugs can regulate the activation of microglia and have a certain clinical or preventive effect. Ulinastain (UTI) is a protease inhibitor, which has been proved to have a clear anti-inflammatory and antioxidant effect in the brain and many other organ systems. It has a regulation on the process of microglia activation. The effect has not been confirmed. This article will discuss some problems based on this problem.
[Objective]
By using lipopolysaccharide (LPS) to build a stereoscopic model of microglia activation, the effect of UTI therapy on the activation of microglia and the effects on the learning and memory function were observed and the signal pathway mechanism was further explored.
[method]
1, the rat model of microglia activation was established by injecting LPS into rats.
2, we use LPS to stimulate BV2 cells to establish microglia models in vitro.
3, Morris Water Maze (MWM) was used to observe the changes of learning and memory function in rats. Morris.
4, the morphological changes of microglia were observed by immunohistochemistry.
5, ELISA was used to detect the release of TNF- alpha and IL-1 beta.
6, Western Blot detected the changes in the related signaling pathways.
[results]
1, LPS induced microglia cell activation model was established.
In the experimental part of the body, after LPS intraperitoneal injection, the results of Morris water maze decreased in rats. Immunohistochemical staining could observe a large number of activated microglia in the hippocampus of the brain. The ELISA detection of brain tissue suggested that the content of TNF- A and IL-1 beta was increased. In vitro, LPS stimulation could increase the release of TNF- A and IL-1 beta in BV2 cells.
2, the inhibitory effect of UTI on microglia activation induced by LPS.
In the body test part, UTI of 100kU/kg could improve the decrease of Morris water maze caused by LPS injection. The results of immunohistochemistry showed that the activation of microglia decreased, and the result of ELISA suggested that the content of TNF- alpha and IL-1 beta decreased. In vitro experimental part, BV2 cells treated by UTI were reduced by LPS stimulated TNF- and IL-1 beta.
3, the signal transduction mechanism of UTI inhibiting LPS induced microglial activation.
Both in vitro and in vivo experiments showed that the activation of MAPKs and JAK-STAT pathways in hippocampal and BV2 cells was up-regulated by LPS stimulation, and their activity decreased after UTI treatment. By inhibiting the expression of partial MAPKs pathway, the activity of STAT3 induced by LPS could be inhibited.
[Conclusion]
1, LPS can activate microglia, increase the release of inflammatory factors, and affect learning and memory function.
2, UTI inhibits the activation of microglia induced by LPS, thereby reducing the release of inflammatory factors and improving learning and memory function.
3, the inhibitory effect of UTI on the activation of microglia caused by LPS may be achieved by inhibiting the MAPKs and JAK-STAT pathways. In this process, there is a link between the MAPKs and JAK-STAT pathways, and MAPKs is involved in the regulation of STAT3 phosphorylation.
To sum up, UTI has a certain protective effect on the nervous system, and is expected to be used as a new drug for the treatment and prevention of neurodegenerative diseases.
【學(xué)位授予單位】：第四軍醫(yī)大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2012
【分類(lèi)號(hào)】：R741;G804.2

【共引文獻(xiàn)】

相關(guān)期刊論文 前10條

1 周虎田,徐如祥,姜曉丹,徐宗俊,蔡穎謙,杜謀選,鄒雨汐,鄧鎮(zhèn),秦玲莎;IL-6促進(jìn)骨髓源性神經(jīng)干細(xì)胞增殖的實(shí)驗(yàn)研究[J];中華神經(jīng)醫(yī)學(xué)雜志;2005年06期

2 吳軍;王愛(ài)桃;閔U

本文編號(hào)：2096989