本文關(guān)鍵詞：TREM2在小膠質(zhì)細(xì)胞介導(dǎo)的帕金森病神經(jīng)炎癥中的作用研究　出處：《南方醫(yī)科大學(xué)》2016年碩士論文　論文類型：學(xué)位論文

  更多相關(guān)文章： 帕金森病 神經(jīng)炎癥 輕度認(rèn)知功能障礙 TREM2 小膠質(zhì)細(xì)胞 慢病毒轉(zhuǎn)染

【摘要】：帕金森病(Parkinson's disease, PD)是常見的中樞神經(jīng)系統(tǒng)退行性疾病,由于早期黑質(zhì)致密部的多巴胺(dopamine, DA)能神經(jīng)元死亡而出現(xiàn)多巴胺耗竭,導(dǎo)致出現(xiàn)以運動遲緩、靜止性震顫、肌強(qiáng)直等為特征的運動障礙。PD也會出現(xiàn)很多非運動癥狀,如輕度認(rèn)知功能障礙(Mild cognitive impairment in Parkisnon's disease, PD-MCI)、癡呆、情緒和精神障礙等。中腦黑質(zhì)DA能神經(jīng)元缺失、α-突觸核蛋白(a-synuclein)沉積和路易小體形成是PD的主要病理特征。目前PD的治療是以增加DA濃度及直接的DA受體激動劑為主的癥狀性治療。疾病修飾治療可減緩神經(jīng)變性或遏制疾病的進(jìn)展,但目前PD的疾病修飾治療仍有待進(jìn)一步探索。因此,闡明PD的發(fā)病機(jī)制將有助于疾病修飾治療的發(fā)展。由于大部分PD患者為散發(fā)性,很可能由遺傳因素和環(huán)境因素相互作用而導(dǎo)致。自從20年前在PD患者中首次報道小膠質(zhì)細(xì)胞活化介導(dǎo)的神經(jīng)炎癥,PD神經(jīng)炎癥成為了研究的焦點。眾所周知,小膠質(zhì)細(xì)胞同時具有細(xì)胞毒性及細(xì)胞保護(hù)性功能。小膠質(zhì)細(xì)胞既可活化為致炎的M1型(或過度活化型),也可活化為抗炎的M2型。M1型小膠質(zhì)細(xì)胞可產(chǎn)生并釋放多種致炎因子,促進(jìn)清除中樞神經(jīng)系統(tǒng)的病原體如伐-突觸核蛋白,但過度活化的小膠質(zhì)細(xì)胞可因產(chǎn)生過多具有神經(jīng)毒性的細(xì)胞因子,反而造成神經(jīng)元損傷繼而加重PD。相反,M2型小膠質(zhì)細(xì)胞可產(chǎn)生抗炎因子、抑制炎癥反應(yīng)、促進(jìn)組織修復(fù)。因此,通過調(diào)控小膠質(zhì)細(xì)胞的亞型分化,促進(jìn)小膠質(zhì)細(xì)胞向抗炎的M2型小膠質(zhì)細(xì)胞轉(zhuǎn)化,將可能為PD提供新的治療策略,但目前小膠質(zhì)細(xì)胞亞型分化的調(diào)控機(jī)制尚未明確。髓樣細(xì)胞表達(dá)的觸發(fā)受體-2 (triggering receptor expressed on myeloid cells 2, TREM2)是表達(dá)于小膠質(zhì)細(xì)胞表面的受體,可減少小膠質(zhì)細(xì)胞介導(dǎo)的炎癥因子導(dǎo)致的促炎反應(yīng),并促進(jìn)小膠質(zhì)細(xì)胞對神經(jīng)元碎片及細(xì)菌的吞噬作用。最近一項大型的GWAS研究表明TREM2基因突變與PD、AD、FTD等發(fā)病風(fēng)險增加有關(guān)。TREM2可能是調(diào)節(jié)PD中小膠質(zhì)細(xì)胞M1/M2亞型平衡的重要因素。本研究將通過臨床研究PD患者外周血中TREM2表達(dá)水平與對照組的差異,并建立PD小鼠模型初步探討TREM2及其下游神經(jīng)炎癥因子的表達(dá)水平,進(jìn)一步通過TREM2-shRNA及wtTREM2慢病毒轉(zhuǎn)染小膠質(zhì)細(xì)胞以使TREM2表達(dá)沉默或過表達(dá),然后分析TREM2表達(dá)水平對小膠質(zhì)細(xì)胞M1/M2亞型分化的影響,以探討TREM2對小膠質(zhì)細(xì)胞亞型分化的調(diào)控作用,以闡明TREM2在小膠質(zhì)細(xì)胞介導(dǎo)的神經(jīng)炎癥中的作用。第一章可溶性TREM2在PD及PD認(rèn)知障礙中的表達(dá)水平研究目的：研究sTREM2在PD患者和正常對照者間的表達(dá)差異,并探討sTREM2對PD認(rèn)知功能障礙的影響。方法：研究納入114例PD患者和120例健康對照者,收集一般臨床資料,采用統(tǒng)一帕金森病評分量表(Unified Parkinson Disease Rating Scale, UPDRS)第1U部分評估PD患者的運動功能,通過MoCA(Montreal Cognitive Assessment, MoCA)量表評估整體認(rèn)知功能、韋氏智力和韋氏記憶量表評估執(zhí)行功能、記憶、注意力及視空間空能。進(jìn)一步根據(jù)PD輕度認(rèn)知功能障礙的診斷標(biāo)準(zhǔn),將PD組分為PD認(rèn)知正常組(PD patients with normal cognition, PD-CN組)和PD輕度認(rèn)知功能障礙組(PD patients with cognitive impairment,PD-MCI組),其中PD-CN組66例,PD-MCI組48例。并采集PD組及對照組的外周血,用ELISA法檢測外周血中sTREM2的表達(dá)水平,分析各組外周血中sTREM2表達(dá)水平的差異,并進(jìn)一步探討sTREM2在PD認(rèn)知功能中的作用。結(jié)果：①114例PD思看中,PD-MCI思者48例,占42.11%。PD-MCI組的受教育水平較PD-CN組低(P0.05)；運動功能(UPDRS-III評分、H-Y分級)及整體認(rèn)知功能(MMSE評分及MoCA評分)均較PD-CN組更差,差異有統(tǒng)計學(xué)意義(P0.05)；②PD患者外周血中sTREM2表達(dá)水平顯著高于健康對照組,差異有統(tǒng)計學(xué)意義(P0.001)；③健康對照組、PD-CN組和PD-MCI組三組間外周血sTREM2表達(dá)水平有顯著性差異(P0.001), PD-MCI組患者外周血中的sTREM2表達(dá)水平顯著高于PD-CN組,差異有統(tǒng)計學(xué)意義(P0.001)；④PD患者外周血中的sTREM2水平與MoCA評分呈中度負(fù)相關(guān)(r=-0.464,P0.001);⑤PD患者外周血中的sTREM2水平升高與受教育年限水平低(P=0.023)、相似性量表分(P=-0.015)及再認(rèn)量表分(P=-0.018)下降相關(guān)。結(jié)論：PD患者外周血的sTREM2表達(dá)水平升高與PD患者受教育年限低、全面認(rèn)知功能下降、執(zhí)行功能及記憶功能下降有關(guān),外周血的sTREM2表達(dá)水平可能有助于監(jiān)測PD及PD認(rèn)知障礙的病情進(jìn)展,表明TREM2可能參與了PD及PD認(rèn)知功能障礙的發(fā)生發(fā)展,但內(nèi)在機(jī)制仍有待進(jìn)一步研究。第二章TREM2對MPTP誘導(dǎo)的PD小鼠模型神經(jīng)炎癥的影響目的：分析MPTP誘導(dǎo)的PD小鼠模型和正常對照小鼠腦組織中TREM2、促炎因子及抗炎因子的表達(dá)水平差異,探討TREM2在PD小鼠神經(jīng)炎癥中的作用。方法：通過C57BL/6J小鼠小劑量長期腹腔注射MPTP和丙磺舒建立PD慢性模型,另設(shè)丙磺舒對照組和生理鹽水對照組。建立PD模型后,取各組小鼠中腦、皮層、海馬、紋狀體等部位腦組織,通過酪氨酸羥化酶(tyrosine hydroxylase, TH)染色觀察中腦DA能神經(jīng)元死亡情況,并采用熒光定量PCR方法檢測：rREM2及炎癥因子IL-6、IL-1β、TNF-a基因的mRNA表達(dá)水平,進(jìn)一步通過western blot檢測TREM2及抗炎因子Arginase-1蛋白的表達(dá)水平,最后應(yīng)用單因素方差分析比較三組間DA能神經(jīng)元缺失、TREM2、炎癥因子及抗炎因子的表達(dá)水平差異。結(jié)果：①MPTP模型組中TH染色陽性DA能神經(jīng)元數(shù)目較丙磺舒對照組及生理鹽水對照組顯著減少(P0.001),MPTP組小鼠中腦TH+神經(jīng)元數(shù)目較生理鹽水對照組減少39.7%：②MPTP模型組、丙磺舒對照組及生理鹽水對照組之間中腦及皮層中TREM2、 IL-1β、IL-6, TNF-a基因的mRNA相對表達(dá)水平有顯著性差異(P0.05),其中MPTP組表達(dá)水平最高,差異有統(tǒng)計學(xué)意義(P0.05)；③MPTP模型組、丙磺舒對照組及生理鹽水對照組三組間中腦、皮層、紋狀體及海馬組織中TREM2及抗炎因子Arginase-1蛋白的表達(dá)水平有顯著性差異(P0.05),其中MPTP組表達(dá)水平高于丙磺舒對照組及生理鹽水對照組,差異有統(tǒng)計學(xué)意義(P0.05)；結(jié)論：MPTP誘導(dǎo)的PD小鼠模型中腦、紋狀體、海馬及皮層等部位腦組織均出現(xiàn)TREM2及抗炎因子Arginase-1蛋白表達(dá)增加,伴有中腦DA能神經(jīng)元缺失及顯著的神經(jīng)炎癥,提示TREM2與PD中腦DA能神經(jīng)元缺失及神經(jīng)炎癥有密切的聯(lián)系,可能是PD出現(xiàn)神經(jīng)炎癥的情況下為了阻止炎癥反應(yīng)而出現(xiàn)的代償性升高,但其確切機(jī)制有待進(jìn)一步闡明。第三章TREM2對小膠質(zhì)細(xì)胞亞型分化的調(diào)節(jié)作用目的：初步探討TREM2是否在小膠質(zhì)細(xì)胞M1/M2亞型分化中發(fā)揮調(diào)控作用。方法：選用BV2細(xì)胞作為小膠質(zhì)細(xì)胞模型,用TREM2-shRNA'慢病毒感染小膠質(zhì)細(xì)胞使TREM2沉默表達(dá),wtTREM2'慢病毒使小膠質(zhì)細(xì)胞過表達(dá)TREM2,并分別設(shè)陰性對照病毒感染組,感染96小時后通過免疫熒光、熒光定量PCR及western blot驗證感染效率。各組慢病毒感染后根據(jù)加入M1型小膠質(zhì)細(xì)胞誘導(dǎo)劑(IL-4+IL-13)或M2型小膠質(zhì)細(xì)胞誘導(dǎo)劑(LPS+IFN-y)的不同,再將小膠質(zhì)細(xì)胞分為3組：對照組、IL-4 (1 Ong/ml)+IL-13(10ng/ml)組、LPS(100ng/ml)) +IFN-y (1 Ong/ml)組。感染96小時后各組分別加入相應(yīng)濃度的M1/M2型誘導(dǎo)劑,24小時后通過免疫熒光、western blot及熒光定量PCR檢測小膠質(zhì)細(xì)胞上TREM2及抗炎因子Arginase-1的表達(dá)水平,并采用Griess法檢測炎癥因子NO的表達(dá)水平。結(jié)果：①TREM2-shRNA慢病毒及scramble TREM2陰性對照病毒、wtTREM2慢病毒及vector陰性對照病毒感染小膠質(zhì)細(xì)胞后,免疫熒光結(jié)果表明TREM2-shRNA'隉病毒及scramble TREM2陰性對照病毒、vector陰性對照病毒在小膠質(zhì)細(xì)胞中的感染效率均達(dá)90%以上,wtTREM2慢病毒的感染效率達(dá)80%以上,可保證后續(xù)研究的進(jìn)行；②慢病毒感染小膠質(zhì)細(xì)胞后,TREM2-shRNA組TREM2及M2型小膠質(zhì)細(xì)胞標(biāo)志物Arginase-1蛋白的表達(dá)水平較scramble TREM2組顯著下降(P0.05),相反,wtTREM2組TREM2及Arginase-1蛋白的表達(dá)水平較vector組顯著升高(P0.05)：③慢病毒感染小膠質(zhì)細(xì)胞并利用M1/M2型小膠質(zhì)細(xì)胞誘導(dǎo)劑后,IL-4+IL-13(M2型小膠質(zhì)細(xì)胞誘導(dǎo)劑)組的TREM2及M2型小膠質(zhì)細(xì)胞標(biāo)志物Arginase-1表達(dá)水平較對照組升高,M1型小膠質(zhì)細(xì)胞標(biāo)志物NO表達(dá)減少(P0.05),而LPS+IFN-γ(M1型小膠質(zhì)細(xì)胞誘導(dǎo)劑)組恰好相反,TREM2及Arginase-1表達(dá)水平較對照組降低,NO表達(dá)增加(P0.05),表明小膠質(zhì)細(xì)胞在M1型/M2型誘導(dǎo)劑下出現(xiàn)了亞型轉(zhuǎn)化；④沉默TREM2表達(dá)后,TREM2及M2型小膠質(zhì)細(xì)胞標(biāo)志物Arginase-1表達(dá)水平顯著下降,而使用M2型小膠質(zhì)細(xì)胞誘導(dǎo)劑IL-4/IL-13可誘導(dǎo)其表達(dá)增加；相反,TREM2過表達(dá)后,TREM2及Arginase-1表達(dá)水平顯著增加,而使用M1型小膠質(zhì)細(xì)胞誘導(dǎo)劑LPS/IFN-γ可誘導(dǎo)其表達(dá)下降；表明TREM2可促進(jìn)小膠質(zhì)細(xì)胞從M1型向M2型轉(zhuǎn)化。結(jié)論：(1)在沒有使用M1型誘導(dǎo)劑的情況下,TREM2沉默表達(dá)即可減弱小膠質(zhì)細(xì)胞向M2型分化的功能,表明TREM2在M2型小膠質(zhì)細(xì)胞分化中發(fā)揮著重要的作用,可能參與了PD神經(jīng)炎癥的發(fā)病機(jī)制；(2) TREM2沉默表達(dá)可抑制小膠質(zhì)細(xì)胞向M2型分化而加重M1型小膠質(zhì)細(xì)胞介導(dǎo)的炎癥反應(yīng),相反,TREM2過表達(dá)可誘導(dǎo)小膠質(zhì)細(xì)胞向M2型轉(zhuǎn)化,抑制M1型小膠質(zhì)細(xì)胞介導(dǎo)的炎癥反應(yīng)并產(chǎn)生抗炎因子,表明TREM2在PD神經(jīng)炎癥中發(fā)揮保護(hù)作用,但TREM2如何調(diào)節(jié)小膠質(zhì)細(xì)胞M1/M2亞型分化的具體分子機(jī)制有待進(jìn)一步闡明。
[Abstract]:Parkinson's disease (Parkinson's disease PD) is a common neurodegenerative disease of the central nervous system, due to the early substantia nigra pars compacta (dopamine, DA) to the death of neurons and dopamine depletion, resulting in bradykinesia, resting tremor, rigidity and other characteristics of movement disorders.PD will also be a lot of non motor symptoms such as, mild cognitive impairment (Mild cognitive impairment in Parkisnon's disease, PD-MCI), dementia, emotional and mental disorders. The substantia nigra DA neurons lack of alpha synuclein (a-synuclein) deposition and Louis body formation is the main pathological features of PD. The PD treatment is symptomatic treatment to increase the concentration of DA and direct DA receptor agonist based. Progress of disease modifying therapy can alleviate nerve degeneration or prevent disease, but the modified PD disease treatment remains to be further explored for. This, to elucidate the mechanism of PD will contribute to the development of disease modifying therapy. Because most PD patients are sporadic, probably by the interaction of genetic and environmental factors caused. Since 20 years ago in PD were reported for the first time in the activation of microglia mediated neuroinflammation, PD nerve inflammation has become a research focus. As everyone knows, microglia also have cell toxicity and cell protective function. Microglia can be activated as M1 type of inflammatory (or excessive activation), can also be activated as anti-inflammatory M2 type.M1 type of microglia can produce and release a variety of inflammatory factors, promote the removal of the central nervous system the pathogens such as cutting - synuclein, but excessive activation of microglial cells by cytokines produced more neurotoxic, but cause neuronal injury and aggravation of PD. instead of M2 type of microglia can produce anti-inflammatory Factor, inhibit inflammation, promote tissue repair. Therefore, the subtype differentiation and regulation of microglial cells, promote the transformation of microglia to anti-inflammatory M2 microglial cells may provide a new therapeutic strategy for PD, but the regulation mechanism of microglia subtype differentiation is not clear. The expression of myeloid cell triggering receptor (triggering receptor expressed -2 on myeloid cells 2, TREM2) is expressed in microglia cell surface receptors, proinflammatory response can reduce inflammatory factors lead to microglia mediated phagocytosis, and promote microglia to neurons and bacterial debris. A recent study showed that large GWAS PD and AD, TREM2 gene mutation, FTD increased risk of.TREM2 may be an important factor in the regulation of microglia in PD M1/M2 subtype balance. This will be through the clinical study of PD in peripheral blood of patients with TREM The difference between the 2 expression levels compared with the control group, and the establishment of a mouse model of PD to investigate the expression level of TREM2 and its downstream nerve inflammation factor, further TREM2-shRNA and wtTREM2 lentiviral transfection of microglia by silencing TREM2 expression or overexpression, and then analyze the influence of TREM2 expression level on microglia M1/M2 subtype differentiation, to to discuss the effects of TREM2 on microglia subtype differentiation regulation, to elucidate the role of TREM2 in microglia mediated neuroinflammation. Objective to study the expression of in the first chapter of soluble TREM2 in PD and PD cognitive impairment: a study of sTREM2 in the differential expression of PD between patients and normal controls, and to investigate the effect of sTREM2 PD of cognitive dysfunction. Methods: the study included 114 PD patients and 120 healthy controls, collected clinical data, using the unified Parkinson's Disease Rating Scale (Unified Parkinson Disease Rating Scale, UPDRS) assessment of motor function in patients with PD and part 1U, through MoCA (Montreal Cognitive Assessment, MoCA) scale to assess the overall cognitive function, intelligence and Wechsler Memory Scale assessment of executive function, memory, attention and visual space can be empty. Further according to the PD diagnostic criteria of mild cognitive impairment. The PD group was divided into PD normal group (PD patients with cognitive normal cognition, PD-CN group) and PD group (PD patients MCI with cognitive impairment, PD-MCI group), including 66 cases of PD-CN group, PD-MCI group of 48 cases. Peripheral blood samples were collected in PD group and control group, the expression level of sTREM2 detection blood samples were analyzed by ELISA method. The expression of sTREM2 in peripheral blood of different levels, and to further explore the role of sTREM2 in PD cognitive function. Results: 114 cases of PD, 48 cases of fancy, in patients with PD-MCI, accounted for 42.11%.PD-MCI group The education level was lower than that of PD-CN group (P0.05); motor function (UPDRS-III score, H-Y grade) and overall cognitive function (MMSE score and MoCA score) were compared with those in PD-CN group, the difference was statistically significant (P0.05); the PD in peripheral blood of patients with the expression level of sTREM2 was significantly higher than that in healthy controls, there are statistically significant difference (P0.001); the healthy control group, PD-CN group and PD-MCI group between the three groups of peripheral blood sTREM2 expression level had significant difference (P0.001), the expression level of PD-MCI group in peripheral blood of patients with sTREM2 was significantly higher than that of PD-CN group, the difference was statistically significant (P0.001); the patients with PD the blood sTREM2 level and MoCA score were negatively correlated (r=-0.464, P0.001); the PD in peripheral blood of patients with elevated levels of sTREM2 and low level of Education (P=0.023), similarity scale (P=-0.015) and recognition scale (P=-0.018) decreased. Conclusion: Patients with PD The high expression level of peripheral blood sTREM2 in patients with PD years of education is low, overall decline in cognitive function, executive function and memory function decline, progress may help monitor PD and cognitive impairment of PD disease of peripheral blood sTREM2 expression showed that TREM2 may be involved in the development of cognitive dysfunction in PD and PD. But the internal mechanism still need further research. The second chapter TREM2 on MPTP induced mouse model of PD nerve inflammation Objective: TREM2 analysis of PD mice model induced by MPTP and normal mouse brain tissue, inflammatory cytokines and anti-inflammatory due to the expression level of sub differences, to explore the role of TREM2 in PD mice. Methods in neuroinflammation PD: to establish a model of chronic C57BL/6J mice by long-term low-dose intraperitoneal injection of MPTP and probenecid, another probenecid control group and saline control group. After the establishment of PD model, the mice in each group, The cerebral cortex, hippocampus, striatum and other parts of the brain, by tyrosine hydroxylase (tyrosine hydroxylase, TH) staining observed in the midbrain DA neurons death, and using fluorescent quantitative PCR method to detect rREM2 and inflammatory cytokines IL-6, IL-1 beta, TNF-a mRNA gene expression level, further through the expression level of Western blot detection of TREM2 and anti-inflammatory factor Arginase-1 protein, single factor variance analysis finally the application of comparison between the three groups of DA neurons lack TREM2, expression of inflammatory and anti-inflammatory factors. Results: the MPTP in the model group TH staining DA positive neurons compared with the number of probenecid control group and saline control group significantly decreased (P0.001), the number of midbrain TH+ neurons. Mice in group MPTP compared with the saline control group reduced 39.7%: the MPTP model group, probenecid between the control group and saline control group of midbrain and cortical layer TREM2, IL-1 beta, I L-6, the relative expression of TNF-a gene mRNA level had significant difference (P0.05), in MPTP group, the highest expression level, the difference was statistically significant (P0.05); the MPTP model group, probenecid control group and saline control group between the three groups in the midbrain, cortex, striatum and hippocampus tissue in the expression level of TREM2 and anti-inflammatory factor Arginase-1 the protein had significant difference (P0.05), the group MPTP expression level was higher than probenecid control group and saline control group, the difference was statistically significant (P0.05); conclusion: mesencephalic PD mice model induced by MPTP, striatum, cerebral cortex and hippocampus and other parts are TREM2 and anti-inflammatory factor Arginase-1 protein expression increased with midbrain DA neurons, and lack of significant nerve inflammation, suggesting that TREM2 and PD in DA is closely related to neuronal loss and nerve inflammation, PD may be the case for nerve inflammation Compensatory to prevent inflammation and increased, but the exact mechanism remains to be clarified. The third chapter TREM2 regulation of microglia subtype differentiation Objective: To investigate whether TREM2 play a regulatory role in microglia M1/M2 subtype differentiation. Methods: using BV2 cells as microglia with TREM2-shRNA'model. Slow virus infection of microglia TREM2 to silence the expression of TREM2 over expression of microglia wtTREM2' lentivirus, and negative control virus infection group, 96 h after infection by immunofluorescence and fluorescence quantitative PCR and Western blot to verify the efficiency of infection. The infected according to the M1 type of microglia induced by agent (IL-4+IL-13) or type M2 microglia inducer (LPS+IFN-y) is different, the microglial cells were divided into 3 groups: control group, IL-4 (1 Ong/ml) +IL-13 (10ng/ml) group, LPS (100ng/m L)) +IFN-y (1 Ong/ml) group. After 96 hours of infection in each group were added to the corresponding M1/M2 type concentration of inducer, after 24 hours by immunofluorescence, the expression level of Western blot and fluorescence quantitative PCR detection of microglia on TREM2 and anti-inflammatory factor Arginase-1, and the expression level of Griess was used to detect the inflammatory factor NO. Results: TREM2-shRNA lentivirus and scramble TREM2 negative control virus, wtTREM2 vector lentivirus and negative control virus infection of microglia after immunofluorescence showed that invasion of TREM2-shRNA'virus and scramble virus vector TREM2 negative control, negative control efficiency of infection in microglia are more than 90%, wtTREM2 lentiviral infection efficiency more than 80%, can ensure the follow-up research; the lentivirus infected microglia after TREM2-shRNA, group TREM2 and M2 microglia marker Arginase-1 egg The expression level of scramble TREM2 than the white group decreased significantly (P0.05), on the contrary, the expression level of TREM2 and Arginase-1 protein in wtTREM2 group were higher than those of group vector (P0.05): the lentivirus infected microglia and microglia induced by M1/M2, IL-4+IL-13 (M2 microglia inducer) group TREM2 and M2 type microglia marker expression level of Arginase-1 was higher than the control group, the expression of type M1 microglia marker NO (P0.05), and LPS+IFN- (gamma M1 microglia inducer) Group on the contrary, the expression level of TREM2 and Arginase-1 were lower than the control group, the increased expression of NO (P0.05). That microglia in M1 type /M2 type inducer appeared under the subtype transformation; the expression of TREM2 silencing and M2 type TREM2, microglia marker Arginase-1 expression level was significantly decreased, and the use of M2 type of microglia induced by IL-4/IL-13 Can induce its expression increase; conversely, overexpression of TREM2, the expression level of TREM2 and Arginase-1 increased significantly, while the use of type M1 microglia LPS/IFN- inducer can induce its expression decreased; showed that TREM2 can promote the transformation of microglia from M1 type to M2 type. Conclusion: (1) without the use of M1 type induction agent, TREM2 silencing can weaken microglial cells to differentiate into type M2 functions show that TREM2 plays an important role in the differentiation of M2 in microglia, may be involved in the pathogenesis of PD nerve inflammation; (2) TREM2 silencing could inhibit the expression of microglia to M2 differentiation and inflammation reaction type M1 microglia mediated instead, overexpression of TREM2 can be transformed to M2 induced microglial cells, inhibiting the inflammatory reaction of M1 microglia mediated and produce anti-inflammatory factors, suggest that TREM2 play a protective PD neural inflammation However, the specific molecular mechanism of how TREM2 regulates the differentiation of M1/M2 subtypes in microglia remains to be further elucidated.

【學(xué)位授予單位】：南方醫(yī)科大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2016
【分類號】：R742.5

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