本文選題：神經退行性病變 + 小膠質細胞�。� 參考：《第四軍醫(yī)大學》2012年碩士論文

【摘要】：【研究背景】 神經退行性病變是一類年齡相關的以神經元功能和結構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，臨床癥狀根據損傷的不同類型表現(xiàn)出不同的行為、精神、認知、運動性障礙。在病理學上，炎癥與氧化應激對神經退行性病變有十分重要的作用。 小膠質細胞作為神經系統(tǒng)的固有免疫單位，和神經退行性病變有非常重要的聯(lián)系。靜息的小膠質細胞對于神經系統(tǒng)有支持、監(jiān)視、修復等功能；小膠質細胞對腦內環(huán)境變化十分敏感，一些刺激能使小膠質細胞活化，小膠質細胞活化可以產生很多炎癥趨化因子和自由基，這些物質大部分具有細胞毒性，蓄積過高可以使神經元退化、變性、死亡，進一步會導致神經退行性病變。多種信號通路都參與了小膠質細胞的活化過程，如JAK-STAT、MAPK等，且不同的通路之間互相聯(lián)系，互相影響，信號通路調控對于調節(jié)小膠質細胞狀態(tài)具有重要意義。因此找到抑制小膠質細胞的活化的方法和途徑在預防和治療神經退行性病變中有重要的意義。以小膠質細胞作為神經退行性病變的治療靶點已經成為一種可行的方式，，目前已證實多種藥物均能調控小膠質細胞的活化，且具有一定的臨床治療或預防作用。烏司他丁（Ulinastain，UTI）是一種蛋白酶抑制劑，在腦及其他多種器官系統(tǒng)中均被證實具有明確的抗炎抗氧化作用，它對于小膠質細胞活化的過程是否有調控作用還未被證實，本文將基于此問題展開一些討論。 【目的】 通過利用脂多糖（lipopolysaccharide，LPS）建立體內外小膠質細胞活化的模型，觀察UTI治療對于小膠質細胞活化的作用和對學習記憶功能的影響。并進一步探討其中的信號通路機制。 【方法】 1、利用LPS注射大鼠建立小膠質細胞活化的體內模型。 2、利用LPS刺激BV2細胞建立小膠質細胞的體外模型。 3、 Morris水迷宮（Morris Water Maze，MWM）觀察大鼠學習記憶功能的變化。 4、免疫組織化學觀察小膠質細胞形態(tài)學的變化。 5、 ELISA檢測TNF-α和IL-1β的釋放。 6、 Western Blot檢測相關信號通路的變化。 【結果】 1、 LPS誘導小膠質細胞細胞活化模型的建立 體內實驗部分，LPS腹腔注射后，大鼠的Morris水迷宮成績下降，免疫組織化學可觀察到腦內海馬區(qū)大量活化的小膠質細胞，腦組織ELISA檢測提示TNF-α和IL-1β的含量增高。體外實驗部分，LPS刺激可以使BV2細胞TNF-α和IL-1β的釋放增高。 2、 UTI對LPS誘導的小膠質細胞活化的抑制效應 體內試驗部分，100kU/kg的UTI對于LPS注射引起的Morris水迷宮成績下降有改善作用，免疫組織化學結果顯示小膠質細胞的活化減少，ELISA結果提示TNF-α和IL-1β含量下降。體外實驗部分，UTI處理的BV2細胞在LPS刺激下TNF-α和IL-1β的釋放減少。 3、 UTI抑制LPS誘導的小膠質細胞活化的信號轉導機制 體內外實驗均發(fā)現(xiàn)，在LPS的刺激可誘導海馬組織和BV2細胞的MAPKs和JAK-STAT通路活性上調，在UTI處理后他們的活性下降。通過抑制部分MAPKs通路的表達可以抑制LPS誘導的STAT3活性的升高。 【結論】 1、 LPS可以使小膠質細胞活化，炎癥因子釋放增多，并影響學習記憶功能。 2、 UTI對LPS引起的小膠質細胞活化有抑制作用，從而降低炎癥因子的釋放，改善學習記憶功能。 3、 UTI對LPS引起的小膠質細胞活化的抑制作用可能是通過抑制MAPKs和JAK-STAT通路實現(xiàn)的。在這個過程中，MAPKs和JAK-STAT通路存在聯(lián)系，MAPKs參與調控STAT3的磷酸化。 綜上所述，UTI對于神經系統(tǒng)有一定的保護作用，可望作為一種新的藥物用于神經退行性病變的治療和預防。
[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.
【學位授予單位】：第四軍醫(yī)大學
【學位級別】：碩士
【學位授予年份】：2012
【分類號】：R741;G804.2

【共引文獻】

相關期刊論文 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促進骨髓源性神經干細胞增殖的實驗研究[J];中華神經醫(yī)學雜志;2005年06期

2 吳軍;王愛桃;閔U

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