本文選題：系統(tǒng)性紅斑狼瘡 + 紅細胞凋亡��； 參考：《安徽醫(yī)科大學》2017年碩士論文

【摘要】：研究背景 系統(tǒng)性紅斑狼瘡(SLE)患者隨病情進展可出現(xiàn)不同程度的貧血癥狀,貧血率可高達70%以上,慢性病性貧血最常見。SLE患者因伴有自由基代謝障礙,活性氧類聚集從而形成機體過氧化狀態(tài)。研究證實紅細胞在氧化壓力、能量供給不足、感染等因素作用下凋亡率增加。探討紅細胞凋亡損傷可能是對研究SLE患者貧血病因的科學補充。目的 研究系統(tǒng)性紅斑狼瘡(SLE)貧血患者紅細胞凋亡損傷,并探討其可能的發(fā)生機制。方法 本研究分別以SLE患者和健康獻血員作為實驗組和對照組,分為紅細胞實驗、血漿孵育紅細胞實驗和血漿抗氧化能力實驗三部分。紅細胞實驗為研究兩組對象紅細胞凋亡損傷。分別收集20例兩組對象的新鮮全血,測量血細胞參數(shù)。常規(guī)分離出紅細胞,制成一定濃度的紅細胞懸液。經(jīng)Annexin V-FITC熒光染色,利用流式細胞儀檢測細胞凋亡率及細胞體積大小,同時用激光共聚焦顯微鏡觀察凋亡細胞的形態(tài);經(jīng)熒光探針Fluo-3/AM和DCFH-DA標記,利用流式細胞儀檢測細胞內(nèi)鈣離子濃度和胞內(nèi)活性氧類含量;熒光標記二抗-流式法檢測胞內(nèi)神經(jīng)酰胺含量。分別比較兩組各項指標差異。血漿孵育紅細胞實驗為研究SLE患者血漿對紅細胞調(diào)亡損傷的影響,分別收集20例兩組對象新鮮全血,分離出的血漿用以孵育健康人O型紅細胞。24小時后用相同方法檢測兩組紅細胞凋亡率及體積、胞內(nèi)鈣離子濃度、神經(jīng)酰胺和活性氧類含量,比較兩組差異。另外利用酶標儀測量比較兩組血漿總抗氧化能力差異。結(jié)果 實驗組血紅蛋白含量及血細胞比容均明顯低于正常對照組,差異有統(tǒng)計學意義(P0.001);紅細胞實驗中,實驗組細胞凋亡率、胞內(nèi)鈣離子濃度顯著高于對照組,差異有統(tǒng)計學意義(P0.001);實驗組胞內(nèi)活性氧類含量明顯高于對照組(P0.01);相比健康獻血員,患者紅細胞體積縮小,差異有統(tǒng)計學意義(P0.01);兩組細胞胞內(nèi)神經(jīng)酰胺含量無明顯差異;激光共聚焦熒光顯微鏡下可以觀察到凋亡紅細胞膜上的FITC熒光信號。血漿孵育紅細胞實驗中,實驗組細胞凋亡率、胞內(nèi)鈣離子濃度顯著高于對照組,差異有統(tǒng)計學意義(P0.001,P0.05);細胞大小、胞內(nèi)神經(jīng)酰胺含量以及活性氧類均無明顯差異。兩組血漿總抗氧化能力也無顯著差異。結(jié)論 紅細胞凋亡增加是引起SLE患者貧血的原因之一。胞內(nèi)鈣離子濃度增加以及細胞自身活性氧類代謝障礙可能是紅細胞凋亡的發(fā)生機制。
[Abstract]:Background: patients with systemic lupus erythematosus (SLEs) may develop anemia symptoms of varying degrees with the progression of the disease, the anemia rate can be as high as more than 70%. The most common chronic anemia. SLE patients with free radical metabolic disorders. Reactive oxygen species accumulate to form a state of peroxidation. The results showed that the apoptosis rate of erythrocytes increased under the action of oxidative pressure, insufficient energy supply and infection. To explore the damage of erythrocyte apoptosis may be a scientific supplement to study the etiology of anemia in patients with SLE. Objective to study the damage of erythrocyte apoptosis in anemia patients with systemic lupus erythematosus (SLEA) and its possible mechanism. Methods in this study, SLE patients and healthy blood donors were divided into three parts: erythrocyte experiment, plasma incubating erythrocyte experiment and plasma antioxidant capacity experiment. Erythrocyte apoptosis injury was studied in two groups. Fresh whole blood was collected from 20 cases of two groups and blood cell parameters were measured. Red blood cells were routinely separated and made into a certain concentration of red blood cell suspension. The apoptosis rate and cell volume were detected by flow cytometry by Annexin V-FITC fluorescence staining, and the morphology of apoptotic cells was observed by confocal laser microscope. The apoptotic cells were labeled with fluorescence probe Fluo-3/AM and DCFH-DA. Intracellular calcium concentration and intracellular reactive oxygen species were detected by flow cytometry and ceramide content was detected by fluorescence labeled second antibody flow cytometry. The difference of each index between the two groups was compared. In order to study the effect of plasma on erythrocyte apoptosis injury in patients with SLE, fresh whole blood samples were collected from 20 patients in two groups. The isolated plasma was used to incubate healthy human type O red blood cells for 24 hours. The apoptosis rate and volume of erythrocytes, intracellular calcium concentration, ceramide and reactive oxygen species contents in the two groups were measured by the same method, and the differences were compared between the two groups. In addition, the difference of plasma total antioxidant capacity between the two groups was measured by enzyme labeling instrument. Results the hemoglobin content and blood cell volume in the experimental group were significantly lower than those in the normal control group (P 0.001), the apoptosis rate and intracellular calcium concentration in the experimental group were significantly higher than those in the control group. The content of reactive oxygen species in the experimental group was significantly higher than that in the control group (P 0.01), the volume of erythrocytes in the patients was smaller than that in the healthy blood donors, and the difference was statistically significant (P 0.01), and there was no significant difference in the content of ceramide between the two groups. The FITC fluorescence signals on the membrane of apoptotic erythrocytes can be observed under laser confocal fluorescence microscope. The apoptosis rate and intracellular calcium concentration in the experimental group were significantly higher than those in the control group (P 0.001 P 0.05), but there was no significant difference in cell size, ceramide content and reactive oxygen species. There was no significant difference in total antioxidant capacity between the two groups. Conclusion the increase of erythrocyte apoptosis is one of the causes of anemia in SLE patients. The increase of intracellular Ca ~ (2 +) concentration and the metabolic disturbance of reactive oxygen species may be the mechanism of erythrocyte apoptosis.
【學位授予單位】：安徽醫(yī)科大學
【學位級別】：碩士
【學位授予年份】：2017
【分類號】：R593.241;R446.11

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相關(guān)期刊論文 前7條

1 Junichi Fujii;Toshihiro Kurahashi;Tasuku Konno;Takujiro Homma;Yoshihito Iuchi;;Oxidative stress as a potential causal factor for autoimmune hemolytic anemia and systemic lupus erythematosus[J];World Journal of Nephrology;2015年02期

2 趙永琴;林s，

本文編號：1934181