兰州大学机构库 >学院待认领
香烟烟雾提取物对慢性阻塞性肺疾病慢患者单核细胞源性巨噬细胞吞噬功能及氧化损伤的影响
Alternative TitleEffects of Cigarette Smoke Extract on Phagocytosis of Monocyte-derived Macrophage and Oxidative Damage in COPD Patients
庞琪
Thesis Advisor刘晓菊
2013-05-10
Degree Grantor兰州大学
Place of Conferral兰州
Degree Name硕士
Keyword香烟烟雾提取物 慢性阻塞性肺疾病 单核细胞源性巨噬细胞 吞噬能力 氧化应激
Abstract目的 探讨香烟烟雾提取物(CSE)对慢性阻塞性肺疾病(COPD)患者单核细胞源性巨噬细胞(MDM)吞噬功能的影响及氧化损伤。 方法 选择COPD患者32例,健康体检者32例作为研究对象。分离外周血单个核细胞,收集贴壁的单核细胞,体外培养并诱导产生单核细胞源性巨噬细胞(MDM)。根据实验要求将COPD患者的MDM分为空白对照组(A组)、CSE干预组(B组);健康体检者的MDM也分为空白对照组(C组)及CSE干预组(D组)。采用MTT比色法检测CSE对MDM活性的影响;流式细胞术结合激光扫描共聚焦显微镜检测各组细胞吞噬荧光素异硫氰酸酯标记的大肠杆菌(FITC-E.coli)的吞噬能力;分光光度计检测CSE干预前后细胞上清液中总抗氧化能力(TAC)、丙二醛(MDA)及谷胱甘肽过氧化物酶(GSH-PX)的水平。 结果(1)随着CSE作用时间及浓度的增加,细胞存活率下降。(2)流式细胞术显示:A组吞噬FITC-E.coli的平均荧光强度(MFI)(20.22±2.22)低于C组(56.90±6.68)(P<0.01);B组的MFI(7.65 ±0.70)较A组下降(P<0.01);D组MFI(47.95±5.35)较C组下降(P<0.01);B组较A组MFI下降的程度(61.92±4.01%)较D组较C组下降程度(15.23±9.16%)更为明显(P<0.01)。(3)激光扫描共聚焦显微镜检测可观察到COPD患者MDM吞噬FITC-E.coli的能力较健康体检者明显降低,CSE对两组MDM干预后其吞噬能力均有所下降,A组吞噬FITC-E.coli的荧光吸光度值( 51.43±5.85)低于C组( 87.33±7.29)(P<0.01);B组的(14.11 ±3.36)较A组下降(P<0.01);D组(69.72±5.97)较C组下降(P<0.01);B组较A组下降的程度( 71.81±8.79%)较D组较C组下降程度( 20.06±8.20%)更为明显(P<0.01);(4)A组TAC、GSH-PX水平 (4.06±0.23,47.10±4.13)U/ml低于C组 (5.14 ±0.29,88.40±2.31)U/ml,MDA水平( 4.75±0.53)nmol/ml高于C组(2.10±0.28)nmol/ml(P均<0.01);B组( 3.07±0..32, 26.75±6.19)U/ml较A组TAC、GSH-PX水平下降(P均<0.01),MDA水平(7.72±0.85)升高(P<0.01);D组( 4.47±0.36、72.23±5.14)U/ml较C组TAC、GSH-PX水平下降(P均<0.01),MDA水平( 2.95±0.56)nmol/ml升高(P<0.01);B组较A组TAC、GSH-PX下降程度(24.51±6.71%、43.40±11.91%)较D组较C组下降程度(13.05±5.13%,18.21±5.79%)更明显(P均<0.01),B组较A组MDA的升高程度(38.20±5.58%)较D组较C组升高程度(26.17.±1.76%)明显(P<0.05)。(5)相关性分析显示:基础状态下COPD患者吞噬FITC-E.coli的MFI与TAC、GSH-PX成正相关(r值分别为0.523和0.818,P分别<0.05 ,P<0.01),与MDA成负相关(r值为0.501,P<0.05),TAC与GSH-PX成正相关(r值为0.615,P<0.01),MDA与TAC、GSH-PX成负相关(r值分别为0.596和0.615,P均<0.05)。CSE干预后COPD患者吞噬 FITC-E.coli的MFI与TAC、GSH-PX成正相关(r值分别为0.704和0.716,P均<0.01),与MDA成负相关(r值为0.522,P<0.05),TAC与GSH-PX成正相关(r值为0.714,P<0.01),MDA与TAC、GSH-PX成负相关(r值分别为0.712和0.532,P分别<0.01 ,<0.05)。 结论 CSE浓度增加,作用时间延长,对MDM的活性影响作用增大,呈剂量-效应关系和时间效应关系;COPD患者MDM的吞噬FITC-E.coli能力较健康者明...
Other AbstractObjective To explore the effccts of CSE on phagocytosis function of monocyte-derived macrophage in COPD patients and oxidative damage. Methods The peripheral blood mononuclear cells of 32 COPD patients and 32 healthy people were isolated. Adherent monocytes were collected from the cells, cultured in vitro and induced monocyte-derived macrophages(MDM) .The MDM from COPD patients were divided into 2groups: control group (group A), CSE treatment group (group B).The MDM from healthy controls were also divided into 2groups: control group (group C), CSE treatment group (group D). Toxin function of CSE on MDM was measured by MTT. Flow cytometry combined with laser scanning confocal microscopyto scan the phagocytic activity of Escherichia coli (E. coli) labeled by FITC among groups.The contents of TAC,MDA and GSH-PX in the cell cultured supernatants were tested by Spectrophotometer . Results (1) The longer time and higher concentration is used, the lower MDM survival rate.(2) Flow cytometry suggest that compared with MFI in group C (56.90±6.68) with E.d might be the reason that MDMrease, , the MFI in group A (20.22±2.22) and group D (47.95±5.35) decreased significantly (P<0.01);compared with MFI in group A, the data in group B (7.65 ±0.70) decreased significantly (P<0.01). The decreased rang of MFI between group A and group B (61.92±4.01%) was significantly higher than group C and group D (15.23±9.16%) (P<0.01).(3) Using laser under confocal microscope conducted that MDM Phagocytosis of E.coli labeled by FITC (FITC-E.coli) of COPD patients was lower than healthy controls, and with CSE treatments, MDM Phagocytosis of all groups were reduced,and compared with the fluorescence absorbance values in group C ( 87.33±7.2) with E.d might be the reason that MDMrease, , the fluorescence absorbance values in group A ( 51.43±5.85) and group D ( 69.72±5.97) decreased significantly (P<0.01);compared with the fluorescence absorbance values in group A, the data in group B (14.11 ±3.36) decreased significantly (P<0.01). The decreased rang of the fluorescence absorbance values between group A and group B (71.81±8.79%) was significantly higher than group C and group D (20.06±8.20%) (P<0.01).(4)The TAC and GSH-PX levels were lower in group A (4.06±0.23,47.10±4.13) U/ml than that in group C (5.14 ±0.29,88.40±2.31) U/ml (P all < 0.01), MDA level was significantly higher in group A( 4.75±0.53)nmol/ml than that in group C (2.10±0.28)nmol/ml.TAC and GSH-...
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Language中文
Document Type学位论文
Identifierhttps://ir.lzu.edu.cn/handle/262010/222509
Collection学院待认领
Affiliation临床医学院
Recommended Citation
GB/T 7714
庞琪. 香烟烟雾提取物对慢性阻塞性肺疾病慢患者单核细胞源性巨噬细胞吞噬功能及氧化损伤的影响[D]. 兰州. 兰州大学,2013.
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