【摘要】：目的:1、研究細胞微粒(Microparticles,MPs)血小板微粒(Platelets microparticles,PMPs)、CD41a+CD62P+PMPs、粒細胞微粒(Granulocyte microparticles,GMPs)、CD4+T細胞微粒(CD4 positive T cells microparticles,CD4+T-LMPs)、CD8+T細胞微粒(CD8 positive T cells microparticles,CD8+T-LMPs)和B細胞微粒(B-cell microparticles,B-LMPs)在強直性脊柱炎(Ankylosing Spondylitis,AS)患者和正常人中的變化,觀察AS患者MPs、PMP、CD41a+CD62P+PMPs、GMPs、CD4+T-LMPs、CD8+T-LMPs和B-LMPs水平與臨床炎癥指標(biāo)的關(guān)系。2、探討AS患者MPs對人臍靜脈內(nèi)皮細胞(human umbilical vein endothelial cell,HUVEC)的細胞存活率影響。進一步分析AS患者血清刺激形成的PMPs對HUVEC細胞間粘附分子-1(Intercellular adhesion molecule-1,ICAM-1)、血管細胞粘附分子-1(Vascular cell adhesion molecule-1,VCAM-1)和內(nèi)皮型一氧化氮合酶(Endothelial nitric oxide synthase,e NOS)基因表達的影響。方法:收集47例AS患者外周血,所有患者均符合1984年修訂的AS紐約標(biāo)準(zhǔn),同時收集15例來院體檢的健康人外周血作為對照組。(1)收集47例AS患者的空腹靜脈血并檢測血沉(ESR)、C反應(yīng)蛋白(CRP)水平、評定BASDAI評分。(2)采用流式細胞術(shù)(Flow cytometry,FCM)檢測AS患者和HC組MPs、PMPs、GMPs、B-LMPs、CD4+T-LMPs、CD8+T-LMPs和CD41a+CD62P+PMPs表達水平并分析AS患者MPs、PMPs、GMPs、B-LMPs、CD4+T-LMPs、CD8+T-LMPs和CD41a+CD62P+PMPs數(shù)量與臨床炎癥指標(biāo)的相關(guān)性。(3)CCK8法檢測HUVEC與MPs共孵育后的細胞存活率。(4)RT-PCR檢測HUVEC與AS患者和HC刺激形成PMPs共孵育24 h后ICAM-1、VCAM-1和e NOSm RNA水平表達。結(jié)果:(1)AS患者MPs、PMPs、CD41a+CD62P+PMPs和GMPs數(shù)量顯著高于健康對照組(p0.05)。AS患者B-LMPs,CD4+T-LMPs、CD8+T-LMPs數(shù)量與健康對照組相比,差異無統(tǒng)計學(xué)意義(p0.05)。(2)AS患者MPs、PMPs、CD41a+CD62P+PMPs、GMPs、B-LMPs、CD4+T-LMPs和CD8+T-LMPs數(shù)量與ESR,CRP無相關(guān)性。(3)AS患者MPs與BASDAI呈正相關(guān)(r=0.5548,P=0.0004)、PMPs與BASDAI呈正相關(guān)(r=0.5423,P=0.0006)、CD41a+CD62P+PMPs、GMPs與BASDAI無相關(guān)性(p0.05)。(4)AS患者與HC組MPs與HUVEC共培養(yǎng)24小時后,AS患者MPs能顯著降低HUVEC細胞存活率(p0.01)。(5)AS與HC組血清刺激形成的PMPs與HUVEC細胞共培養(yǎng)24小時后,AS組、HC組和空白組之間e NOS、VCAM-1m RNA表達無明顯統(tǒng)計學(xué)差異(p0.05)。而AS組ICAM-1m RNA表達顯著高于空白組和HC組,差異具有統(tǒng)計學(xué)意義(p0.01)。結(jié)論:AS患者MPs、PMPs、GMPs和CD41a+CD62P+PMP表達增加。并且AS患者MPs,PMPs數(shù)量與BASDAI呈正相關(guān),提示MPs,PMPs可能參與AS發(fā)病。此外AS患者MPs能夠明顯降低HUVEC細胞存活率,AS患者MPs可能損傷內(nèi)皮細胞而參與心血管功能調(diào)控。此外,AS患者血清刺激產(chǎn)生的PMPs能夠上調(diào)ICAM-1m RNA表達水平,提示MPs中的PMPs可能通過調(diào)節(jié)內(nèi)皮細胞相關(guān)功能基因損傷內(nèi)皮細胞,這可能在一定程度上導(dǎo)致AS患者心血管風(fēng)險增加。
[Abstract]:Objective: 1. To study the cellular microparticles (Microparticles,MPs), platelet microparticles (Platelets microparticles,PMPs), CD41a CD62P PMPs, granulocyte particles (Granulocyte microparticles,GMPs), CD4 T cell particles (CD4 positive T cells microparticles,CD4 T-LMPs), CD8 T cell particles (CD8 positive T cells microparticles,). The changes of CD8 T-LMPs and B cell microparticles (B-cell microparticles,B-LMPs) in patients with ankylosing spondylitis (Ankylosing Spondylitis,AS) and normal controls were observed. To investigate the effect of MPs on the survival rate of human umbilical vein endothelial cells (human umbilical vein endothelial cell,HUVEC) in patients with AS. 2. The relationship between the levels of CD8 T-LMPs and B-LMPs and clinical inflammatory indexes. Further analysis of HUVEC intercellular adhesion molecule-1 (Intercellular adhesion molecule-1,ICAM-1 (ICAM-1 (Intercellular adhesion molecule-1,ICAM-1), vascular cell adhesion molecule-1 (Vascular cell adhesion molecule-1,VCAM-1 (VCAM-1 (Vascular cell adhesion molecule-1,VCAM-1) and endothelial nitric oxide synthase (Endothelial nitric oxide synthase, (Enos) induced by serum-stimulated PMPs in AS patients was carried out. The effect of e NOS) gene expression. Methods: peripheral blood samples were collected from 47 patients with AS, all of whom met the AS New York standard revised in 1984. At the same time, the peripheral blood of 15 healthy people were collected as control group. (1) fasting venous blood was collected from 47 patients with AS and erythrocyte sedimentation rate (ESR) (ESR), C reactive protein (CRP) level was measured. (2) flow cytometry (Flow cytometry,FCM) was used to detect the expression of MPs,PMPs,GMPs,B-LMPs,CD4 T cells, CD 8 T-LMPs and CD41a CD62P PMPs in patients with AS and HC and to analyze MPs,PMPs,GMPs,B-LMPs, in patients with AS. (2) BASDAI scores were evaluated by flow cytometry (Flow cytometry,FCM). CD4 T\ x {e16c} LMPs, The relationship between the number of CD8 T-LMPs and CD41a CD62P PMPs and clinical inflammatory markers. (3) the survival rate of HUVEC-MPs co-incubated with HUVEC was detected by CCK8. (4) ICAM-1, was detected by RT-PCR after PMPs was co-incubated with AS and HC for 24 h. VCAM-1 and e NOSm RNA were expressed at the level. Results: (1) the number of MPs,PMPs,CD41a CD62P PMPs and GMPs in AS patients was significantly higher than that in healthy controls (p0.05). There was no significant difference (p0.05). (2) between the number of MPs,PMPs,CD41a CD62P PMPs,GMPs,B-LMPs,CD4 T-LMPs and CD8 T-LMPs and ESR,CRP in AS patients. (3) there was a positive correlation between MPs and BASDAI in AS patients (r = 0.555 8, P < 0.0004). There was a positive correlation between PMPs and BASDAI (r = 0.5423, P = 0.0006), but there was no correlation between CD41a CD62P PMPs,GMPs and BASDAI (p0.05). (4). After 24 hours of co-culture of MPs and HUVEC between AS patients and HC group, MPs and HUVEC were co-cultured for 24 hours. The survival rate of HUVEC cells was significantly decreased by MPs in AS patients (p0.01). (5). After 24 hours of co-culture of PMPs and HUVEC cells induced by AS and serum stimulation in HC group, e NOS, was observed between AS group, HC group and blank group. There was no significant difference in the expression of VCAM-1m RNA (p0.05). The expression of ICAM-1m RNA in AS group was significantly higher than that in blank group and HC group (p0.01). Conclusion: the expression of MPs,PMPs,GMPs and CD41a CD62P PMP increased in AS patients. There was a positive correlation between the number of MPs,PMPs and BASDAI in patients with AS, suggesting that MPs,PMPs may be involved in the pathogenesis of AS. In addition, MPs can significantly reduce the survival rate of HUVEC cells in AS patients, and MPs in AS patients may damage endothelial cells and participate in cardiovascular function regulation. In addition, PMPs produced by serum stimulation in patients with AS can up-regulate the expression of ICAM-1m RNA, suggesting that PMPs in MPs may damage endothelial cells by regulating endothelial cell-related functional genes. This may, to some extent, lead to increased cardiovascular risk in patients with AS.
【學(xué)位授予單位】：西南醫(yī)科大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2017
【分類號】：R593.23

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本文編號：2469594