Ang-Ⅱ-p22phox-ROS通路在川崎病发病机制中的作用

发布时间：2018-03-16 20:09

本文选题：川崎病　切入点：冠脉损害　出处：《川北医学院》2017年硕士论文　论文类型：学位论文

【摘要】：第一部分Ang-Ⅱ、p22phox与川崎病及其冠脉损害相关性的研究目的:探索Ang-Ⅱ、p22phox二者在川崎病及冠脉损害中的作用。方法:1.酶联免疫吸附实验(enzyme-linked immunosorbent assay,ELISA)检测51(15例冠脉损害,36例冠脉正常)例川崎病患儿及28名正常体检儿童血浆Ang-Ⅱ。2.实施荧光定量聚合酶链反应法(real-time polymerase chain reaction,RT-PCR)检测40例川崎病患儿(15例冠脉损害,25例冠脉正常)和20名正常患儿体检外周血单个核细胞p22phox m RNA表达。结果:1.川崎病血浆Ang-Ⅱ浓度冠脉损害组(Coronary artery lesions,CALs)与无冠脉损害组(Non Coronary artery lesions,NCALs)表达无明显差异(P0.05),CALs及NCALs均显著高于正常组(P0.01)。2.川崎病外周血单个核细胞p22phox m RNA表达水平CALs与无冠脉损害组NCALs表达无明显差异(P0.05),CALs与NCALs均高于正常组(P0.05)。3.川崎病外周血单个核细胞p22phox m RNA表达水平与血浆Ang-Ⅱ浓度呈正相关关系(r=0.358,P=0.023)。结论:Ang-Ⅱ、p22phox可能参与了川崎病的发病机制,是否在川崎病冠脉损害中起作用有待进一步研究。第二部分静脉用丙种球蛋白治疗对川崎病Ang-Ⅱ及p22phoxm RNA表达的影响目的:探索静脉丙种球蛋白(intravenous immunoglobulin,IVIG)治疗的作用机制,以及IVIG治疗对川崎病Ang-Ⅱ及p22phox m RNA表达的影响。方法:1.ELISA检测51例川崎病患儿治疗前、治疗后及28名健康儿童血浆Ang-Ⅱ浓度。2.RT-PCR检测40例川崎病患儿IVIG治疗前、治疗后(15例冠脉损害和25例无冠脉损害)及20名健康儿童外周血单个核细胞p22phox m RNA表达水平。结果:1.患儿血浆Ang-Ⅱ浓度,治疗前明显高于治疗后及正常组(P0.01),治疗后明显高于正常组(P0.01)。2.川崎病患儿外周血单个核细胞p22phox m RNA表达水平治疗前高于治疗后(P0.05),明显高于正常组(P0.01),治疗后与正常组比较无明显差异(P0.05)。结论:IVIG治疗对Ang-Ⅱ及p22phox m RNA表达在急性期均有影响。第三部分川崎病患儿血清对体外培养的血管内皮细胞的损伤作用目的:探讨川崎病患儿治疗前后血清干预对体外培养的血管内皮细胞的作用及临床意义方法:1.分别以川崎病患儿治疗前、治疗后和健康儿童血清(体积分数均为10%)处理人脐静脉内皮细胞(HUVEC)(川崎病患儿治疗前、后血清组n=36;健康儿童血清组n=19)24小时。2.倒置相差显微镜观察细胞形态;3.流式细胞术检测细胞凋亡率;4.酶联免疫ELISA法检测10例患儿培养细胞上清液中血管紧张素Ⅱ(AngⅡ)的质量浓度;5.Real-Time PCR检测内皮细胞p22phox m RNA表达。结果:1.与健康血清组和空白对照组比较,治疗前及治疗后血清组HUVEC的细胞数量明显减少,细胞核固缩、边缘化,胞质浓缩。2.治疗前血清组细胞凋亡率为(17.48±1.14)%,治疗后血清组细胞凋亡率为(8.33±0.79)%,健康血清组细胞凋亡率为(6.09±0.45)%,空白对照组细胞凋亡率为(4.22±0.39)%。3.治疗前、后血清组细胞培养上清中AngⅡ的质量浓度无明显差异(P0.05)。4.与治疗后血清组及健康血清组比较,治疗前血清组p22phox m RNA表达增高(P0.01),治疗后血清组与正常组表达无明显差异(P0.05)。结论:川崎病患儿治疗前血清干预使HUVEC细胞形态发生改变,抑制细胞增殖并促进细胞凋亡,p22phoxm RNA表达增高可能导致内皮细胞功能受损。
[Abstract]:The first part of Ang-, p22phox and Kawasaki disease and coronary artery lesion Objective: To explore the correlation between Ang- II and p22phox two role in disease and coronary artery lesion in Kawasaki. Methods: 1. enzyme-linked immunosorbent assay (enzyme-linked immunosorbent, assay, ELISA) was detected in 51 (15 cases of coronary artery lesion, 36 cases with normal coronary arteries) with Kawasaki sick children and 28 normal children on plasma Ang-.2. fluorescence quantitative polymerase chain reaction (real-time polymerase chain reaction, RT-PCR) were detected in 40 patients (Kawasaki 15 cases of coronary artery damage, 25 cases of normal coronary artery) and expression of p22phox in mononuclear cells of M RNA blood of 20 normal children peripheral examination. Results: 1. Kawasaki disease plasma Ang- II coronary artery lesion group (Coronary artery lesions, CALs) and non coronary artery lesion group (Non Coronary, artery lesions, NCALs) showed no significant difference (P0.05), CALs and NCALs were significantly higher than normal group (P 0.01) Kawasaki.2. disease p22phox in peripheral blood mononuclear cells m RNA expression level of CALs and coronary artery lesion group NCALs showed no significant difference (P0.05), CALs and NCALs were higher than the normal group (P0.05).3. Kawasaki disease p22phox in peripheral blood mononuclear cells m RNA expression level correlated with plasma Ang- II concentration was positively related (r=0.358, P=0.023). Conclusion: Ang- II, p22phox may be involved in pathogenesis of the disease is in Kawasaki, Kawasaki plays a role in coronary artery lesion disease needs further study. The second part of intravenous immunoglobulin therapy effect on the expression of Kawasaki Ang- II and p22phoxm disease RNA Objective: To explore the intravenous immunoglobulin (intravenous immunoglobulin. IVIG) the mechanism of therapeutic effect, and the effect of IVIG treatment on the expression of Kawasaki Ang- II and p22phox m RNA disease. Methods: with Kawasaki 1.ELISA detection in the treatment of 51 cases of disease before and after treatment and 28 healthy children on plasma Ang- concentration Kawasaki.2.RT-PCR IVIG disease detection treatment in 40 cases before and after treatment (15 cases of coronary artery lesion and 25 cases without coronary artery lesions) the level of p22phox in peripheral blood mononuclear cells m expression of RNA and 20 healthy children. Results: 1. patients with plasma Ang- II concentration before treatment was significantly higher than that after treatment and the normal control group (P0.01), after treatment was significantly higher than the normal group (P0.01) p22phox in peripheral blood mononuclear cells m RNA expression of.2. in children with Kawasaki disease levels before treatment than after treatment (P0.05), significantly higher than the normal group (P0.01), after treatment and control group had no significant difference (P0.05). Conclusion: the expression of IVIG in acute treatment effect all the stages of Ang- II and p22phox m RNA. The third part Kawasaki injury disease serum on cultured vascular endothelial cells Objective: To investigate the serum before and after treatment in children with Kawasaki disease intervention on vascular endothelial cells cultured in vitro and its clinical significance 1. respectively in the treatment of children with Kawasaki disease before after treatment and serum of healthy children (volume 10%) treated human umbilical vein endothelial cells (HUVEC) (treated with Kawasaki before disease, serum n=36 group; serum n=19 healthy children group) 24 hours.2. inverted microscope to observe cell morphology; cell apoptosis was detected in 3. flow cytometry detection rate; 4. enzyme-linked immunosorbent ELISA method in 10 cases of vascular cell culture supernatant of angiotensin II (Ang II) mass concentration; the expression of 5.Real-Time PCR p22phox m RNA to detect endothelial cells. Results: 1. with healthy serum group and blank control group, the number of HUVEC cells decreased significantly in serum group before and after treatment, nuclear condensation, marginalization, cytoplasm condensed.2. before treatment serum cell apoptosis rate was (17.48 + 1.14)% after treatment, serum cell apoptosis rate was (8.33 + 0.79)%, healthy blood group apoptosis rate was (6.09 + 0.45)%, Blank control group apoptosis rate was (4.22 + 0.39)%.3. before treatment, serum concentration of Ang II cells in the supernatant had no significant difference (P0.05) after the treatment with.4. serum group and healthy serum group, before treatment serum p22phox m increased expression of RNA (P0.01) after treatment, serum and the normal group showed no significant difference (P0.05). Conclusion: children with Kawasaki disease before treatment serum HUVEC changes in cell morphology, inhibit cell proliferation and promote cell apoptosis, p22phoxm expression of RNA may lead to endothelial cell dysfunction.

【学位授予单位】：川北医学院
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：R725.4

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