中国心血管杂志
中國心血管雜誌
중국심혈관잡지
CHINESE JOURNAL OF CARDIOVASOLOGY
2015年
4期
278-283
,共6页
动脉粥样硬化%糖尿病%脂蛋白,高密度脂蛋白%细胞粘附分子%血清淀粉样蛋白A%载脂蛋白A-Ⅰ
動脈粥樣硬化%糖尿病%脂蛋白,高密度脂蛋白%細胞粘附分子%血清澱粉樣蛋白A%載脂蛋白A-Ⅰ
동맥죽양경화%당뇨병%지단백,고밀도지단백%세포점부분자%혈청정분양단백A%재지단백A-Ⅰ
Atherosclerosis%Diabetes mellitus%Lipoproteins,HDL%Cell adhesion molecules%Serum amyloid A protein%Apolipoprotein A-Ⅰ
目的：比较2型糖尿病和冠心病患者与健康人高密度脂蛋白(HDL)对内皮细胞表达粘附分子的抑制能力,并研究其与 HDL 中血清淀粉样蛋白 A(SAA)含量改变的相关性。方法通过密度梯度离心法从健康人、单纯糖尿病患者、糖尿病合并冠心病患者以及单纯冠心病患者血浆样本中提取 HDL,用酶联免疫吸附法检测 HDL 中 SAA 及载脂蛋白 AⅠ(apoAⅠ)含量。将健康人及不同患者的 HDL 与脐静脉内皮细胞(HUVEC)共同孵育,通过肿瘤坏死因子α刺激,分别与抗人 CD54-PE、CD106-FITC 单克隆抗体及 Calcein-AM 荧光染色剂标记后的 THP-1细胞共同孵育后,流式细胞术检测平均荧光强度获得 HUVEC 细胞间粘附分子1(ICAM-1)、血管细胞粘附分子1(VCAM-1)表达数量及 THP-1细胞粘附数量,从而比较不同组别 HDL 抑制粘附分子表达及抑制细胞粘附的能力。结果健康人群 HDL 中 SAA 含量与单纯糖尿病及单纯冠心病患者 HDL 中 SAA 含量相比均明显降低[(4.89±1.46)ng/μg 比(22.54±7.40)ng/μg 及(13.28±5.05)ng/μg,均为 P ﹤0.05]。健康对照组及各疾病组 HDL 均可使 HUVEC 表达 ICAM-1、VCAM-1水平明显下降,其中健康对照组 HDL 抑制ICAM-1表达能力均明显强于糖尿病组及糖尿病合并冠心病组(17318.60±4364.77比34171.67±13918.07及39633.60±8728.22,均为 P ﹤0.05)。健康对照组 HDL 抑制 VCAM-1表达能力明显强于糖尿病合并冠心病组(244.80±119.55比388.80±75.76,P =0.04)。健康对照组及各疾病组 HDL均可抑制 THP-1细胞粘附 HUVEC 的能力,但健康对照组 HDL 抑制细胞粘附能力明显强于糖尿病组、冠心病组及糖尿病合并冠心病组(503.24±63.50比2918.00±0.00、1510.33±1353.87及4196.50±412.60,均为 P ﹤0.05)。 HDL 中 SAA 的含量与其抗 ICAM-1表达(回归系数为1.57,t =3.22,P ﹤0.01)及 THP-1细胞粘附能力(回归系数为1.16,t =3.05,P =0.01)具有明显相关性。 HDL 中apoAⅠ的含量与其抗 ICAM-1表达能力具有明显相关性(回归系数为-0.35,t =-2.40,P =0.02)。结论糖尿病和冠心病患者 HDL 抑制血管内皮细胞粘附分子表达及抑制单核细胞粘附能力均显著减弱。SAA 含量越多,HDL 抑制粘附能力越弱；apoAⅠ含量越高,HDL 抑制细胞粘附能力越强。
目的：比較2型糖尿病和冠心病患者與健康人高密度脂蛋白(HDL)對內皮細胞錶達粘附分子的抑製能力,併研究其與 HDL 中血清澱粉樣蛋白 A(SAA)含量改變的相關性。方法通過密度梯度離心法從健康人、單純糖尿病患者、糖尿病閤併冠心病患者以及單純冠心病患者血漿樣本中提取 HDL,用酶聯免疫吸附法檢測 HDL 中 SAA 及載脂蛋白 AⅠ(apoAⅠ)含量。將健康人及不同患者的 HDL 與臍靜脈內皮細胞(HUVEC)共同孵育,通過腫瘤壞死因子α刺激,分彆與抗人 CD54-PE、CD106-FITC 單剋隆抗體及 Calcein-AM 熒光染色劑標記後的 THP-1細胞共同孵育後,流式細胞術檢測平均熒光彊度穫得 HUVEC 細胞間粘附分子1(ICAM-1)、血管細胞粘附分子1(VCAM-1)錶達數量及 THP-1細胞粘附數量,從而比較不同組彆 HDL 抑製粘附分子錶達及抑製細胞粘附的能力。結果健康人群 HDL 中 SAA 含量與單純糖尿病及單純冠心病患者 HDL 中 SAA 含量相比均明顯降低[(4.89±1.46)ng/μg 比(22.54±7.40)ng/μg 及(13.28±5.05)ng/μg,均為 P ﹤0.05]。健康對照組及各疾病組 HDL 均可使 HUVEC 錶達 ICAM-1、VCAM-1水平明顯下降,其中健康對照組 HDL 抑製ICAM-1錶達能力均明顯彊于糖尿病組及糖尿病閤併冠心病組(17318.60±4364.77比34171.67±13918.07及39633.60±8728.22,均為 P ﹤0.05)。健康對照組 HDL 抑製 VCAM-1錶達能力明顯彊于糖尿病閤併冠心病組(244.80±119.55比388.80±75.76,P =0.04)。健康對照組及各疾病組 HDL均可抑製 THP-1細胞粘附 HUVEC 的能力,但健康對照組 HDL 抑製細胞粘附能力明顯彊于糖尿病組、冠心病組及糖尿病閤併冠心病組(503.24±63.50比2918.00±0.00、1510.33±1353.87及4196.50±412.60,均為 P ﹤0.05)。 HDL 中 SAA 的含量與其抗 ICAM-1錶達(迴歸繫數為1.57,t =3.22,P ﹤0.01)及 THP-1細胞粘附能力(迴歸繫數為1.16,t =3.05,P =0.01)具有明顯相關性。 HDL 中apoAⅠ的含量與其抗 ICAM-1錶達能力具有明顯相關性(迴歸繫數為-0.35,t =-2.40,P =0.02)。結論糖尿病和冠心病患者 HDL 抑製血管內皮細胞粘附分子錶達及抑製單覈細胞粘附能力均顯著減弱。SAA 含量越多,HDL 抑製粘附能力越弱；apoAⅠ含量越高,HDL 抑製細胞粘附能力越彊。
목적：비교2형당뇨병화관심병환자여건강인고밀도지단백(HDL)대내피세포표체점부분자적억제능력,병연구기여 HDL 중혈청정분양단백 A(SAA)함량개변적상관성。방법통과밀도제도리심법종건강인、단순당뇨병환자、당뇨병합병관심병환자이급단순관심병환자혈장양본중제취 HDL,용매련면역흡부법검측 HDL 중 SAA 급재지단백 AⅠ(apoAⅠ)함량。장건강인급불동환자적 HDL 여제정맥내피세포(HUVEC)공동부육,통과종류배사인자α자격,분별여항인 CD54-PE、CD106-FITC 단극륭항체급 Calcein-AM 형광염색제표기후적 THP-1세포공동부육후,류식세포술검측평균형광강도획득 HUVEC 세포간점부분자1(ICAM-1)、혈관세포점부분자1(VCAM-1)표체수량급 THP-1세포점부수량,종이비교불동조별 HDL 억제점부분자표체급억제세포점부적능력。결과건강인군 HDL 중 SAA 함량여단순당뇨병급단순관심병환자 HDL 중 SAA 함량상비균명현강저[(4.89±1.46)ng/μg 비(22.54±7.40)ng/μg 급(13.28±5.05)ng/μg,균위 P ﹤0.05]。건강대조조급각질병조 HDL 균가사 HUVEC 표체 ICAM-1、VCAM-1수평명현하강,기중건강대조조 HDL 억제ICAM-1표체능력균명현강우당뇨병조급당뇨병합병관심병조(17318.60±4364.77비34171.67±13918.07급39633.60±8728.22,균위 P ﹤0.05)。건강대조조 HDL 억제 VCAM-1표체능력명현강우당뇨병합병관심병조(244.80±119.55비388.80±75.76,P =0.04)。건강대조조급각질병조 HDL균가억제 THP-1세포점부 HUVEC 적능력,단건강대조조 HDL 억제세포점부능력명현강우당뇨병조、관심병조급당뇨병합병관심병조(503.24±63.50비2918.00±0.00、1510.33±1353.87급4196.50±412.60,균위 P ﹤0.05)。 HDL 중 SAA 적함량여기항 ICAM-1표체(회귀계수위1.57,t =3.22,P ﹤0.01)급 THP-1세포점부능력(회귀계수위1.16,t =3.05,P =0.01)구유명현상관성。 HDL 중apoAⅠ적함량여기항 ICAM-1표체능력구유명현상관성(회귀계수위-0.35,t =-2.40,P =0.02)。결론당뇨병화관심병환자 HDL 억제혈관내피세포점부분자표체급억제단핵세포점부능력균현저감약。SAA 함량월다,HDL 억제점부능력월약；apoAⅠ함량월고,HDL 억제세포점부능력월강。
Objective In this case-control study we enrolled healthy subjects and subjects with type 2 diabetes mellitus (DM), coronary heart disease(CHD) only, or both disorders. The objective is to study the inhibition of intracellular adhesion molecule-1 (ICAM-1), vascular cell adhesion molecule-1(VCAM-1) expression and the cell adhesion by high density lipoprotein(HDL) in different groups; and to investigate the relationship between the level of HDL-serum amyloid A ( SAA) and anti-adhesion function. Methods HDLs were isolated from plasma with ultracentrifugation. The levels of lipoprotein-associated SAA and apolipoprotein A-Ⅰ (apoAⅠ) were detected with Enzyme-Linked Immuno Sorbent Assay(ELISA). Human umbilical vein endothelial cells (HUVECs) were incubated with HDLs and stimulated with TNF-α, then labeled with antiCD54-PE, or antiCD106-FITC monoclonal antibodies respectively, then incubated with THP-1 cells, which were labeled by calcein-AM antibody. The expression of cell-surface molecules and the number of adhering THP-1 cells were measured as fluorescence intensity by flow cytometer. Results The levels of HDL-SAA in the healthy controls were lower than patients with DM only and CHD only [(4. 89 ± 1. 46) ng/ μg vs. (22. 54 ± 7. 40) ng/ μg and (13. 28 ± 5. 05) ng/ μg, both P ﹤ 0. 05]. The HDLs of all groups could inhibit the expression of ICAM-1, VCAM-1, while the HDLs obtained from healthy subjects could inhibit the expression of ICAM-1 significantly greater than HDLs obtained from DM and DM + CHD subjects (17 318. 60 ± 4 364. 77 vs. 34 171. 67 ± 13 918. 07 and 39 633. 60 ± 8 728. 22, both P ﹤ 0. 05), and the inhibition of VCAM-1 expression by normal HDLs was significantly greater than HDLs obtained from DM + CHD subjects (244. 80 ± 119. 55 vs. 388. 80 ± 75. 76, P = 0. 04) . The HDLs of all groups could inhibit the adhesion of THP-1. The HDLs from healthy subjects could inhibit the adhesion of THP-1 significantly greater than those from DM, CHD and DM + CHD subjects (503. 24 ± 63. 50 vs. 2 918. 00 ± 0. 00, 1 510. 33 ± 1 353. 87 and 4 196. 50 ± 412. 60, all P ﹤ 0. 05). The level of HDL-SAA was correlated to the level of ICAM-1 expression (the regression coefficient was 1. 57, t = 3. 22, P ﹤ 0. 01) and THP-1 adhesion (the regression coefficient was 1. 16, t = 3. 05, P = 0. 01). In addition, the level of apoAⅠwas reversely correlated with the level of ICAM-1 expression (the regression coefficient was - 0. 35, t = - 2. 40, P = 0. 02) . Conclusions The levels of ICAM-1, VCAM-1 expression and THP-1 adhesion induced by HDL are significantly attenuated in DM subjects and CHD subjects, which are positively correlated with the level of HDL-SAA and reversely correlated with the level of apoAⅠ.
