亲环素A在平滑肌细胞泡沫化过程中表达的变化及氨氯地平的影响
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摘要
一、亲环素A在氧化低密度脂蛋白诱导的平滑肌细胞中表达的变化
目的通过氧化低密度脂蛋白(ox-LDL)诱导的平滑肌细胞,随着ox-LDL浓度和时间的变化,观察亲环素AmRNA和蛋白质的表达改变,为进一步分析亲环素A在ox-LDL诱导的平滑肌细胞中的作用奠定基础。
方法实验采用不同浓度(0,20,40,80,160 mg/L)ox-LDL与本室建立的酶消化法分离的大鼠主动脉血管平滑肌细胞[1],共同孵育72小时,用逆转录聚合酶链反应(RT-PCR)和免疫印迹(Western blot)分别检测亲环素A mRNA和蛋白质的表达改变;然后选择80mg/L ox-LDL与细胞共同孵育不同的时间(0,24,48,72,96小时),同样用RT-PCR和Western blot分别检测亲环素A mRNA和蛋白质的表达改变。结合文献,我们选择80mg/L ox-LDL与细胞共同孵育0,72小时两组,油红O染色观察细胞内脂滴的形成情况,高效液相色谱法检测细胞内胆固醇含量。
结果不同浓度ox-LDL与血管平滑肌细胞共同孵育,随ox-LDL浓度增加,亲环素A mRNA和蛋白质的表达呈现浓度依赖性减弱;80mg/L ox-LDL与细胞共同孵育不同的时间,随ox-LDL孵育时间延长,亲环素A mRNA和蛋白质的表达呈现时间依赖性减弱;差异有显著性(P< 0.05)。油红O染色和高效液相色谱法检测结果显示,80mg/L ox-LDL与细胞共同孵育72小时,细胞胞浆内大量脂滴存在,细胞内胆固醇酯/总胆固醇比值为57.9% (>50%),细胞符合泡沫细胞特征。
结论Ox-LDL可抑制血管平滑肌细胞中亲环素A的表达并诱导泡沫细胞形成。
二、氨氯地平对ox-LDL诱导的血管平滑肌细胞泡沫化过程中亲环素A表达的影响及其对细胞内胆固醇蓄积的作用
目的观察不同浓度(0,0.1,1.0,10.0μM)氨氯地平对ox-LDL诱导的平滑肌细胞泡沫化过程中亲环素A表达的影响及其对胆固醇蓄积的作用,从一个新的角度探讨氨氯地平的抗动脉粥样硬化作用。
方法在第一部分实验基础上,我们选择ox-LDL的处理浓度与时间分别为80 mg/L、72小时,建立血管平滑肌细胞泡沫化模型。实验分为6组:对照组、ox-LDL组、(0.1,1.0,10.0μM)氨氯地平预先处理细胞1小时后加入80 mg/L ox-LDL共同孵育72小时组及氨氯地平(10.0μM)单独处理组,油红O染色观察细胞内脂滴的形成情况,高效液相色谱法检测细胞内胆固醇含量;Western-blot和RT-PCR分别检测亲环素A mRNA和蛋白质的表达改变;进而分析氨氯地平对ox-LDL诱导的平滑肌细胞泡沫化过程中亲环素A表达的影响及其对胆固醇蓄积的作用。
结果用80mg/ L ox-LDL与平滑肌细胞共同孵育72小时,油红O染色显示,ox-LDL组细胞内大量脂滴形成,细胞内胆固醇酯/总胆固醇的比值为57.9%,符合泡沫细胞特征;预先予氨氯地平处理后,随着药物浓度的增加,细胞内脂滴逐渐减少,细胞内胆固醇酯/总胆固醇的比值逐渐降低,至10.0μM氨氯地平处理细胞组效果最明显,为37.8%;RT-PCR和Western-blot检测,发现ox-LDL处理组,细胞内亲环素A的表达明显减少;预处理氨氯地平,随着药物浓度的增加,可使细胞内亲环素A的表达逐渐增加,呈现浓度依赖性,尤其10.0μM氨氯地平预处理细胞组效果最明显,较ox-LDL组有显著性差异,P<0.05。
结论采用氨氯地平预先处理,可上调ox-LDL诱导的平滑肌细胞泡沫化过程中亲环素A的表达,减轻细胞内的胆固醇蓄积,从而延缓或抑制ox-LDL诱导的平滑肌细胞泡沫化过程。
PartⅠ. Change of Cyclophilin A Expression in Smooth Muscle Cells Induced by Ox-LDL
AIM To observe the mRNA and protein expression of Cyclophilin A in smooth muscle cells induced by oxidized low density lipoprotein(ox-LDL)of different concentration and different time and to analyze the effect of Cyclophilin A on smooth muscle cells induced by ox-LDL.
METHODS Rat aortic vascular smooth muscle cells isolated by enzyme digestion applicated generally in our own laboratory were treated with different concentration (0,20,40,80,160 mg/L)of ox-LDL for 72 hours. RT-polymerase chain reaction(RT-PCR)and Western blot analysis were used to detect the change of mRNA and protein expression of Cyclophilin A. Then 80 mg/L ox-LDL was chosen into smooth muscle cells for different tim(e0,24,48,72,96h). RT-PCR and Western blot analysis were also used to detect the change of mRNA and protein expression of Cyclophilin A respectively. Integrated with documents, we chose the cells were treated with 80mg/L ox-LDL for 0 and 72h. Oil Red O Dye experiment showed the lipid droplets in the cells of the two groups. High Performance Liquid Chromatography(HPLC) analysis was peformed the relative value of cellular cholesterol easter and total cholesterol.
RESULTS With the increasing of the concentration of ox-LDL, the mRNA and protein expression of Cyclophilin A in the cells was significantly decreased in a concentration-dependent manner. When 80mg/L ox-LDL was treated with the cells for different time, the expression of Cyclophilin A was also significantly decreased in a time-dependent manner. P< 0.05,there was significant discrepancy. Oil Red O dye showed that there were a lot of lipid droplets accumulated in the smooth muscle cells induced by 80mg/L ox-LDL for 72 hours, the relative value of cellular cholesterol easter and total cholesterol detected by HPLC analysis was 57.9%(>50%). This kind of cells were consistent with the character of the foam cells.
CONCLUSION Ox-LDL can suppress the expression of Cyclophilin A in the smooth muscle cells.
PartⅡ. Function of Amlodipine on Cellular Cholesterol Accumulation in Foam Cells Derived from Smooth Muscle Cells Induced by Ox-LDL and on the Expression of Cyclophilin A
AIM To study the function of Amlodipine of different concentration on cellular cholesterol accumulation in foam cells derived from smooth muscle cells induced by ox-LDL and on the expression of Cyclophilin A to explore the new antiatherosclerotic role of Amlodipine.
METHODS Considering the result above-mentioned,we chose the concentration and time of ox-LDL treated to be 80 mg/L and 72h, at the same time, we constructed the model of foam cells derived from smooth muscle cells induced by ox-LDL.The experiment groups included 6 groups:control group, ox-LDL group, different concentration(0.1,1.0,10.0μM)of Amlodipine pretreated for 1h and treated with 80 mg/L ox-LDL for 72h group and Amlodipine(10.0μM) treated alone without ox-LDL group. Oil Red O Dye experiment showed the lipid droplets in the cells of each group. High Performance Liquid Chromatography(HPLC) analysis was peformed the content of cellular total cholesterol and cholesterol ester. RT-PCR and Western-blot analysis were used to detect the mRNA and protein expression of Cyclophilin A. Then the function of Amlodipine on cellular cholesterol accumulation in foam cells derived from smooth muscle cells induced by ox-LDL and on the expression of Cyclophilin A was analyzed to explore the new antiatherosclerotic role of Amlodipine.
RESULTS Oil Red O dye showed that with the treatment of Amlodipine, the lipid droplets were greatly decreased in a concentration-dependent manner vs 80mg/L ox-LDL for 72 h group. HPLC analysis was peformed the content of cellular total cholesterol and cholesterol ester were both significantly increased with ox-LDL treatment but decreased after Amlodipine treatment. RT-PCR and Western-blot analysis showed that with ox-LDL and Amlodipine treatment, Cyclophilin A expression were greatly increased in a concentration-dependent manner.
CONCLUSION Amlodipine can up-regulate the expression of Cyclophilin A, relieve the cellular cholesterol accumulation and delay the foam cells derived from smooth muscle cells induced by ox-LDL.
目的通过氧化低密度脂蛋白(ox-LDL)诱导的平滑肌细胞,随着ox-LDL浓度和时间的变化,观察亲环素AmRNA和蛋白质的表达改变,为进一步分析亲环素A在ox-LDL诱导的平滑肌细胞中的作用奠定基础。
方法实验采用不同浓度(0,20,40,80,160 mg/L)ox-LDL与本室建立的酶消化法分离的大鼠主动脉血管平滑肌细胞[1],共同孵育72小时,用逆转录聚合酶链反应(RT-PCR)和免疫印迹(Western blot)分别检测亲环素A mRNA和蛋白质的表达改变;然后选择80mg/L ox-LDL与细胞共同孵育不同的时间(0,24,48,72,96小时),同样用RT-PCR和Western blot分别检测亲环素A mRNA和蛋白质的表达改变。结合文献,我们选择80mg/L ox-LDL与细胞共同孵育0,72小时两组,油红O染色观察细胞内脂滴的形成情况,高效液相色谱法检测细胞内胆固醇含量。
结果不同浓度ox-LDL与血管平滑肌细胞共同孵育,随ox-LDL浓度增加,亲环素A mRNA和蛋白质的表达呈现浓度依赖性减弱;80mg/L ox-LDL与细胞共同孵育不同的时间,随ox-LDL孵育时间延长,亲环素A mRNA和蛋白质的表达呈现时间依赖性减弱;差异有显著性(P< 0.05)。油红O染色和高效液相色谱法检测结果显示,80mg/L ox-LDL与细胞共同孵育72小时,细胞胞浆内大量脂滴存在,细胞内胆固醇酯/总胆固醇比值为57.9% (>50%),细胞符合泡沫细胞特征。
结论Ox-LDL可抑制血管平滑肌细胞中亲环素A的表达并诱导泡沫细胞形成。
二、氨氯地平对ox-LDL诱导的血管平滑肌细胞泡沫化过程中亲环素A表达的影响及其对细胞内胆固醇蓄积的作用
目的观察不同浓度(0,0.1,1.0,10.0μM)氨氯地平对ox-LDL诱导的平滑肌细胞泡沫化过程中亲环素A表达的影响及其对胆固醇蓄积的作用,从一个新的角度探讨氨氯地平的抗动脉粥样硬化作用。
方法在第一部分实验基础上,我们选择ox-LDL的处理浓度与时间分别为80 mg/L、72小时,建立血管平滑肌细胞泡沫化模型。实验分为6组:对照组、ox-LDL组、(0.1,1.0,10.0μM)氨氯地平预先处理细胞1小时后加入80 mg/L ox-LDL共同孵育72小时组及氨氯地平(10.0μM)单独处理组,油红O染色观察细胞内脂滴的形成情况,高效液相色谱法检测细胞内胆固醇含量;Western-blot和RT-PCR分别检测亲环素A mRNA和蛋白质的表达改变;进而分析氨氯地平对ox-LDL诱导的平滑肌细胞泡沫化过程中亲环素A表达的影响及其对胆固醇蓄积的作用。
结果用80mg/ L ox-LDL与平滑肌细胞共同孵育72小时,油红O染色显示,ox-LDL组细胞内大量脂滴形成,细胞内胆固醇酯/总胆固醇的比值为57.9%,符合泡沫细胞特征;预先予氨氯地平处理后,随着药物浓度的增加,细胞内脂滴逐渐减少,细胞内胆固醇酯/总胆固醇的比值逐渐降低,至10.0μM氨氯地平处理细胞组效果最明显,为37.8%;RT-PCR和Western-blot检测,发现ox-LDL处理组,细胞内亲环素A的表达明显减少;预处理氨氯地平,随着药物浓度的增加,可使细胞内亲环素A的表达逐渐增加,呈现浓度依赖性,尤其10.0μM氨氯地平预处理细胞组效果最明显,较ox-LDL组有显著性差异,P<0.05。
结论采用氨氯地平预先处理,可上调ox-LDL诱导的平滑肌细胞泡沫化过程中亲环素A的表达,减轻细胞内的胆固醇蓄积,从而延缓或抑制ox-LDL诱导的平滑肌细胞泡沫化过程。
PartⅠ. Change of Cyclophilin A Expression in Smooth Muscle Cells Induced by Ox-LDL
AIM To observe the mRNA and protein expression of Cyclophilin A in smooth muscle cells induced by oxidized low density lipoprotein(ox-LDL)of different concentration and different time and to analyze the effect of Cyclophilin A on smooth muscle cells induced by ox-LDL.
METHODS Rat aortic vascular smooth muscle cells isolated by enzyme digestion applicated generally in our own laboratory were treated with different concentration (0,20,40,80,160 mg/L)of ox-LDL for 72 hours. RT-polymerase chain reaction(RT-PCR)and Western blot analysis were used to detect the change of mRNA and protein expression of Cyclophilin A. Then 80 mg/L ox-LDL was chosen into smooth muscle cells for different tim(e0,24,48,72,96h). RT-PCR and Western blot analysis were also used to detect the change of mRNA and protein expression of Cyclophilin A respectively. Integrated with documents, we chose the cells were treated with 80mg/L ox-LDL for 0 and 72h. Oil Red O Dye experiment showed the lipid droplets in the cells of the two groups. High Performance Liquid Chromatography(HPLC) analysis was peformed the relative value of cellular cholesterol easter and total cholesterol.
RESULTS With the increasing of the concentration of ox-LDL, the mRNA and protein expression of Cyclophilin A in the cells was significantly decreased in a concentration-dependent manner. When 80mg/L ox-LDL was treated with the cells for different time, the expression of Cyclophilin A was also significantly decreased in a time-dependent manner. P< 0.05,there was significant discrepancy. Oil Red O dye showed that there were a lot of lipid droplets accumulated in the smooth muscle cells induced by 80mg/L ox-LDL for 72 hours, the relative value of cellular cholesterol easter and total cholesterol detected by HPLC analysis was 57.9%(>50%). This kind of cells were consistent with the character of the foam cells.
CONCLUSION Ox-LDL can suppress the expression of Cyclophilin A in the smooth muscle cells.
PartⅡ. Function of Amlodipine on Cellular Cholesterol Accumulation in Foam Cells Derived from Smooth Muscle Cells Induced by Ox-LDL and on the Expression of Cyclophilin A
AIM To study the function of Amlodipine of different concentration on cellular cholesterol accumulation in foam cells derived from smooth muscle cells induced by ox-LDL and on the expression of Cyclophilin A to explore the new antiatherosclerotic role of Amlodipine.
METHODS Considering the result above-mentioned,we chose the concentration and time of ox-LDL treated to be 80 mg/L and 72h, at the same time, we constructed the model of foam cells derived from smooth muscle cells induced by ox-LDL.The experiment groups included 6 groups:control group, ox-LDL group, different concentration(0.1,1.0,10.0μM)of Amlodipine pretreated for 1h and treated with 80 mg/L ox-LDL for 72h group and Amlodipine(10.0μM) treated alone without ox-LDL group. Oil Red O Dye experiment showed the lipid droplets in the cells of each group. High Performance Liquid Chromatography(HPLC) analysis was peformed the content of cellular total cholesterol and cholesterol ester. RT-PCR and Western-blot analysis were used to detect the mRNA and protein expression of Cyclophilin A. Then the function of Amlodipine on cellular cholesterol accumulation in foam cells derived from smooth muscle cells induced by ox-LDL and on the expression of Cyclophilin A was analyzed to explore the new antiatherosclerotic role of Amlodipine.
RESULTS Oil Red O dye showed that with the treatment of Amlodipine, the lipid droplets were greatly decreased in a concentration-dependent manner vs 80mg/L ox-LDL for 72 h group. HPLC analysis was peformed the content of cellular total cholesterol and cholesterol ester were both significantly increased with ox-LDL treatment but decreased after Amlodipine treatment. RT-PCR and Western-blot analysis showed that with ox-LDL and Amlodipine treatment, Cyclophilin A expression were greatly increased in a concentration-dependent manner.
CONCLUSION Amlodipine can up-regulate the expression of Cyclophilin A, relieve the cellular cholesterol accumulation and delay the foam cells derived from smooth muscle cells induced by ox-LDL.
引文
[1]涂永生,黄红林,朱炳阳,等. II型胶原酶/弹性蛋白酶消化法培养大鼠血管平滑肌细胞[J].中国动脉粥样硬化杂志, 2001, 9(5): 438-440.
[2]涂玉林,杨小毅,黄红林,等.尼群地平对食饵性家兔动脉粥样硬化的影响Ⅰ、对血浆胆固醇和主动脉病变的作用[J].中国动脉硬化杂志, 1994, 2(4): 153-155.
[3]涂玉林,杨小毅,刘练平,等.尼群地平对食饵性兔动脉粥样硬化的影响Ⅱ、血小板聚集性和血浆脂质过氧化的变化[J].中国动脉硬化杂志, 1995, 3 (4) : 312- 314.
[4]舒春兰,袁中华,雷小勇,等.尼群地平对猪主动脉平滑肌细胞泡沫化的影响[J].衡阳医学院学报, 1998, 26(3): 272, 273, 304.
[5] García Pérez B, Ayala I, Castells MT, et al. Planimetric and histological study of the aortae in atherosclerotic chickens treated with nifedipine, verapamil and diltiazem[J]. Histol Histopathol, 2003, 18(4): 1027- 1033.
[6]刘海宁,张兴华,耿庆信.拉西地平、氨氯地平对人脐静脉内皮细胞间粘附分子1表达的影响[J].中国动脉硬化杂志, 2005, 13(5): 575-578.
[7]于振香,王春艳,陈宇,等.氨氯地平对家兔实验性动脉粥样硬化的防治作用[J].中国老年学杂志, 2003, 10(23): 688-689.
[8] Toba H, Nakagawa Y, Miki S, et al. Calcium channel blockades exhibit anti-inflammatory and antioxidative effects by eugmentation of endothelial nitric oxide synthase and the inhibition of angiotensin converting enzyme in the n(g)-nitro-L-arginine methyl ester-induced hypertensive rat aorta: vasoprotective effects beyond the blood pressure-lowering effects of amlodipine and manidipine [J]. Hypertens Res, 2005, 28 (8): 689-700.
[9] Yoshii T, Iwai M, Li Z, et al. Regression of atherosclerosis by amlodipine via anti-inflammatory and anti-oxidative stress actions[J]. Hypertens Res, 2006, 29(6): 457– 466.
[10] Batova S, DeWever J, Godfraind T, et al. The calcium channel blocker amlodipine promotes the unclamping of eNOS from caveolin in endothelial cells [J]. Cardiovasc Res, 2006, 71(3) : 478-485.
[11] Toba H, Shimizu T, Miki S, et al. Calcium Channel Blockers Reduce AngiotensinII Induced Superoxide Generation and Inhibit Lectin-Like Oxidized Low-Density Lipoprotein Receptor-1 Expression in Endothelial Cells [J]. Hypertens Res, 2006, 29(2): 105-116.
[12] Kataoka C, Egashira K, Ishibashi M, et al. Novel anti-inflammatory actions of amlodipine in a rat model of arteriosclerosis induced by long-term inhibition of nitric oxide synthesis[J]. Am J Physiol Heart Circ Physiol, 2004, 286(2): 768-774.
[13] Ziesche R, Petkov V, Lambers C, et al. The calcium channel blocker amlodipine exerts its antiproliferative action via p21(Waf1/Cip1) gene activation[J]. FASEB J, 2004, 18(13): 1516-1523.
[14]王东,丁华.洛伐他汀、普罗布考对小鼠巨噬细胞源性泡沫细胞内胆固醇含量的影响[J].中国药理学通报, 2004, 20(6): 680-684.
[15]廖端芳,庹勤慧,罗迪贤,等.荷脂细胞胆固醇流出的体系构成与偶联关系[J].南华大学学报:医学版, 2004, 32(3): 347-351.
[16] Yancey PG, Jerome WG. Lysosomal cholesterol derived from mildly oxidized low density lipoprotein is resistant to efflux[J]. J Lipid Res, 2001, 42(3): 317-327.
[17] Dhaliwal BS, Steinbrecher UP. Cholesterol delivered to macrophages by oxidized low density lipoprotein is sequestered in lysosomes and fails to efflux normally[J]. J Lipid Res, 2000, 41(10): 1658-1665.
[18] Gelissen IC, Brown AJ, Mander EL, et al. Sterol efflux is impaired from macrophage foam cells selectively enriched with 7-ketocholesterol[J]. J Biol Chem, 1996, 271(30): 17852-17860.
[19] Young SG, and C. J. Fielding. The ABCs of cholesterol efflux[J]. Nat. Genet, 1999, 22: 316-318.
[20]唐朝克,杨永宗.三磷酸腺苷结合盒转运体A1在THP-1巨噬细胞源性泡沫细胞胆固醇流出中的作用[J].中国病理生理杂志, 2003, 19(8): 1084-1088.
[21]冯翔,凌文华.氧化低密度脂蛋白及溶血卵磷脂对巨噬泡沫细胞胆固醇外流的影响[J].中国病理生理杂志, 2003, 19(9): 1246-1249.
[22]吕湛.胆固醇流出调节蛋白与胆固醇逆向转动和动脉粥样硬化[J].国外医学:生理病理科学与临床分册, 2002, 22(4): 326-328.
[23] Smart EJ, Y.S. Ying, W.C. Donzell, et al. A role for caveolin in transport of cholesterol from endoplasmic reticulum to plasma membrane[J]. J Biol Chem, 1996, 271: 29427-29435.
[24] Uittenbogaard A, Ying Y, Smart EJ. Characterization of a cytosolic heat-shock protein-caveolin chaperone complex. Involvement in cholesterol trafficking[J]. J Biol Chem, 1998, 273(11): 6525-6532.
[25]严鹏科,廖端芳,杨永宗. Caveolin-1表达对血管平滑肌细胞胆固醇逆转运的调节作用[J].中国动脉硬化杂志, 2002, l0(5): 379-383.
[26]王蓉蓉,严鹏科,廖端芳,等.囊泡-囊泡素1在清道夫受体AⅠ转基因小鼠动脉粥样硬化发生中的作用[J].中国动脉粥样硬化杂志, 2002, 10(6): 461-464.
[27] Takahashi N, Hayano T, Suzuki M. Peptidyl-prolyl cis-trans isomerase is the cyclosporinA binding protein cyclophilin[J]. Nature, 1989, 337(6206): 473-475.
[28] Maki K, Ikura T, Hayano T, et al. Effects of proline mutations on the folding of staphylococcal nuclease[J]. Biochemistry, 1999, 38(7): 2213-2223.
[29] Zhao Y, Ke H. Crystal structure implies that cyclophilin predominantly catalyzes the trans to cis isomerization[J]. Biochemistry, 1996, 35(23): 7356-7361.
[30] Jing ZG, Melaragno MG, Liao DF, et al. CyclophilinA is a secreted growth factor induced by oxidative stress[J]. Circ Res, 2000, 87(9): 789-796.
[31] Jin ZG, Lungu AO, Xie L, et al. Cyclophilin A Is a Proinflammatory Cytokine that Activates Endothelial Cells[J]. Arterioscler Thromb Vasc Biol, 2004, 24(7): 1186-1191.
[32] Piotukh K, Gu W, Kofler M, et al. Cyclophilin A binds to linear peptide motifs containing a consensus that is present in many human proteins[J]. J Biol Chem, 2005, 280(25): 23668-23674.
[33]丁振华.低密度脂蛋白的氧化修饰及其理化,生物学性质的改变[J].国外医学:生理病理科学与临床分册, 1994, 14(2): 108-110.
[34]蒋佩,严鹏科,莫中成,等.促进细胞内胆固醇流出抑制单核细胞源性泡沫细胞凋亡[J ].中国病理生理杂志, 2005, 21 (6) : 1041- 1045.
[35]董军,陈文祥,李健斋.高效液相色谱测定微量胆固醇氧化产物[J].生物化学与生物物理进展, 1996, 23: 179-182.
[36] Takahashi N, Hayano T, Suzuki M. Peptidyl-prolyl cis-trans isomerase is the cyclosporinA binding protein cyclophilin[J]. Nature, 1989, 337(6206): 473-475.
[37] Maki K, Ikura T, Hayano T, et al. Effects of proline mutations on the folding of staphylococcal nuclease[J]. Biochemistry, 1999, 38(7): 2213-2223.
[38] Annette Uittenbogaard, Eric J Smart. Palmitoylation of Caveolin-1 is required forcholesterol binding, chaperone complex formation, and rapid transport of cholesterol to caveolae[J]. J Biol Chem, 2000, 75(33): 25595-25599.
[39] Kallen J, Walkinshaw MD. The X-ray structure of a tetrapeptide bound to the active site of human Cyclophilin A[J]. FEBS Lett, 1992, 300(3): 286-290.
[40] Zhao Y, Ke H. Crystal structure implies that cyclophilin predominantly catalyzes the trans to cis isomerization[J]. Biochemistry, 1996, 35(23): 7356-7361.
[41]涂剑,严鹏科,王北冰,等.亲环素A在巨噬细胞荷脂过程中的影响[J ].中国动脉硬化杂志, 2005, 13 (5): 541-544.
[1] Levitsky K, Boersma MD, Ciolli CJ, et al. Exo-Mechanism Proximity- Accelerated Alkylations: Investigations of Linkers, Electrophiles and Surface Mutations in Engineered Cyclophilin-Cyclosporin Systems[J]. Chembiochem, 2005, 6(5): 890-899.
[2] Uittenbogaard A, Everson WV, Matveev SV, et al. Cholesteryl ester is transported from caveolae to internal membranes as part of a caveolin-annexin II lipid-proteincomplex[J]. J Biol Chem, 2002, 277(7):4925-4931.
[3] Piotukh K, Gu W, Kofler M, et al. Cyclophilin A binds to linear peptide motifs containing a consensus that is present in many human proteins[J]. J Biol Chem, 2005, 280(25): 23668-23674.
[4] Jin ZG, Lungu AO, Xie L, et al. Cyclophilin A Is a Proinflammatory Cytokine that Activates Endothelial Cells[J]. Arterioscler Thromb Vasc Biol, 2004, 24(7):1186- 1191.
[5] Jing ZG, Melaragno MG, Liao DF, et al. Cyclophilin A is a secreted growth factor induced by oxidative stress[J]. Circ Res, 2000, 87(9): 789-796.
[6] Bennett MR , Even GI , Schwartz SM, et al. Apoptosis of hunan vascular smooth musde cells derived from noumal vessels and coronary atherosclerotic plaques [ J ] . J Clin Invest, 1995, 95 (5) : 2266– 2274.
[7]唐朝克,杨永宗.三磷酸腺苷结合盒转运体A1在THP-1巨噬细胞源性泡沫细胞胆固醇流出中的作用.中国病理生理杂志, 2003, 19(8): 1084-1088.
[8]冯翔,凌文华.氧化低密度脂蛋白及溶血卵磷脂对巨噬泡沫细胞胆固醇外流的影响.中国病理生理杂志, 2003, 19(9): 1246-1249.
[9]吕湛.胆固醇流出调节蛋白与胆固醇逆向转动和动脉粥样硬化.国外医学:生理病理科学与临床分册, 2002, 22(4): 326-328.
[10]廖端芳,庹勤慧,罗迪贤,等.荷脂细胞胆固醇流出的体系构成与偶联关系.南华大学学报·医学版, 2004, 32(3): 347-351.
[11]Andreea O, Lungu, Zheng - Gen Jin, et al. Cyclosporin A inhibits flow - mediated activation of endothelial nitric-oxide synthase by altering cholesterol content in caveolae[J]. J Biol Chem, 2004, 279: 48794-48800.
[12]Awasthi V, Mandal SK, Papanna V, et al. Modulation of Tissue Factor-Factor VIIa Signaling by Lipid Rafts and Caveolae[J]. Arterioscler Thromb Vasc Biol, 2007, 27(6): 1447-1455.
[13]Covey SD, Brunet RH, Gandhi SG, et al. Cholesterol depletion inhibits fatty acid uptake without affecting CD36 or caveolin-1 distribution in adipocytes[J]. Biochem Biophys Res Commun, 2007, 355(1):67-71.
[14]Theriault Y, Logan TM, Meadows R, et al. Solution structure of the cyclosphorin A/cyclophilin complex by NMR [J]. Nature, 1993, 361: 88-91.
[15]Muruyama T, Frutani M. Archaeal peptidyl prolyl cis-trans isomerases(PPIases)[J].Front Biosci, 2000, 5: 821-836.
[16]Luban J, Bossolt KL, Franke EK, et al. Human immunodeficiency virus type 1 Gag protein binds to cyclophilinsA and B[J]. Cell, 1993, 73(6):1067-1078.
[17]Scholz I, Arvidson B, Huseby D, et al. Virus particle core defects caused by mutations in the human immunodeficiency virus capsid N-terminal domain[J]. J Virol, 2005, 79(3): 1470-1479.
[18]Scholz C, Schaarschmidt P, Engel AM, et al. Functional solubilization of aggregation-prone HIV envelope proteins by covalent fusion with chaperone modules[J]. J Mol Biol, 2005, 345(5):1229-1241.
[19]Chatellard-Gruaz D, Saurat JH, Siegenthaler G. Differential expression of cyclophilin isoforms during keratinocyte differentiation[J]. Biochem J, 1994, 303 (3):863-867.
[20]Watanabe S, Tsuruoka S, Vijayakumar S, et al. Cyclosporin A produces distal renal tubular acidosis by blocking peptidyl prolyl cis-trans isomerase activity of cyclophilin[J]. Am J Physiol Renal Physiol, 2005, 288(1): F40-47.
[21]涂剑,严鹏科,王北冰,等.亲环素A在巨噬细胞荷脂过程中的影响[J].中国动脉硬化杂志, 2005, 13(5): 541-544.
[22]廖端芳,庹勤慧,罗迪贤,等.荷脂细胞胆固醇流出的体系构成与偶联关系.南华大学学报·医学版, 2004, 32(3): 347-351.
[23]廖端芳,严奉祥. Caveolin-1、CyPA与动脉粥样硬化的关系.中国病理生理杂志, 2002, 18(13): 1663-1665.
[24]Zhang YM, Wang KQ, Zhou GM, et al. Endothelin– 1 promoted proliferation of vascular smooth muscle cell through pathway of extracellular signal - regulated kinase and cyclin D1 [J] . Acta Pharmacol Sin, 2003, 24: 563 -568.
[25]Yang H, Li M, Chai H, et al. Effects of cyclophilin A on cell proliferation and gene expressions in human vascular smooth muscular cells and endothelial cells [J]. J Surg Res, 2005, 123(2) :312 - 319.
[26]周四桂,雷小勇,廖端芳.缺氧预适应对缺氧复氧诱导内皮细胞-中性粒细胞黏附的影响[J].中国危重病急救医学, 2003, 15(3): 159-163.
[27]涂光辉,王北冰,严鹏科,等.亲环素A在巨噬细胞荷脂过程中的表达及普罗布考干预的影响[J].中国动脉硬化杂志, 2006, 14(4): 293-296.
[28]Geng YJ, Libby K. Evidence for apoptosis in advanced human[J]. A m J Pathol, 1995, 147 (2) : 251– 266.
[29]Han DK, Haudenschild CC , Hong MK, et al. Evidence for apoptosis in human atherogenesis and in a rat vascular injury model [J]. A m J Pathol, 1995, 147 (2): 267– 277.
[30]Li Z, Zhao X, Bai S, et al. Proteomic identification of cyclophilin A as a potential prognostic factor and therapeutic target in endometrial carcinoma. Mol Cell Proteomics, 2008, 7(10): 1810-1823.
[2]涂玉林,杨小毅,黄红林,等.尼群地平对食饵性家兔动脉粥样硬化的影响Ⅰ、对血浆胆固醇和主动脉病变的作用[J].中国动脉硬化杂志, 1994, 2(4): 153-155.
[3]涂玉林,杨小毅,刘练平,等.尼群地平对食饵性兔动脉粥样硬化的影响Ⅱ、血小板聚集性和血浆脂质过氧化的变化[J].中国动脉硬化杂志, 1995, 3 (4) : 312- 314.
[4]舒春兰,袁中华,雷小勇,等.尼群地平对猪主动脉平滑肌细胞泡沫化的影响[J].衡阳医学院学报, 1998, 26(3): 272, 273, 304.
[5] García Pérez B, Ayala I, Castells MT, et al. Planimetric and histological study of the aortae in atherosclerotic chickens treated with nifedipine, verapamil and diltiazem[J]. Histol Histopathol, 2003, 18(4): 1027- 1033.
[6]刘海宁,张兴华,耿庆信.拉西地平、氨氯地平对人脐静脉内皮细胞间粘附分子1表达的影响[J].中国动脉硬化杂志, 2005, 13(5): 575-578.
[7]于振香,王春艳,陈宇,等.氨氯地平对家兔实验性动脉粥样硬化的防治作用[J].中国老年学杂志, 2003, 10(23): 688-689.
[8] Toba H, Nakagawa Y, Miki S, et al. Calcium channel blockades exhibit anti-inflammatory and antioxidative effects by eugmentation of endothelial nitric oxide synthase and the inhibition of angiotensin converting enzyme in the n(g)-nitro-L-arginine methyl ester-induced hypertensive rat aorta: vasoprotective effects beyond the blood pressure-lowering effects of amlodipine and manidipine [J]. Hypertens Res, 2005, 28 (8): 689-700.
[9] Yoshii T, Iwai M, Li Z, et al. Regression of atherosclerosis by amlodipine via anti-inflammatory and anti-oxidative stress actions[J]. Hypertens Res, 2006, 29(6): 457– 466.
[10] Batova S, DeWever J, Godfraind T, et al. The calcium channel blocker amlodipine promotes the unclamping of eNOS from caveolin in endothelial cells [J]. Cardiovasc Res, 2006, 71(3) : 478-485.
[11] Toba H, Shimizu T, Miki S, et al. Calcium Channel Blockers Reduce AngiotensinII Induced Superoxide Generation and Inhibit Lectin-Like Oxidized Low-Density Lipoprotein Receptor-1 Expression in Endothelial Cells [J]. Hypertens Res, 2006, 29(2): 105-116.
[12] Kataoka C, Egashira K, Ishibashi M, et al. Novel anti-inflammatory actions of amlodipine in a rat model of arteriosclerosis induced by long-term inhibition of nitric oxide synthesis[J]. Am J Physiol Heart Circ Physiol, 2004, 286(2): 768-774.
[13] Ziesche R, Petkov V, Lambers C, et al. The calcium channel blocker amlodipine exerts its antiproliferative action via p21(Waf1/Cip1) gene activation[J]. FASEB J, 2004, 18(13): 1516-1523.
[14]王东,丁华.洛伐他汀、普罗布考对小鼠巨噬细胞源性泡沫细胞内胆固醇含量的影响[J].中国药理学通报, 2004, 20(6): 680-684.
[15]廖端芳,庹勤慧,罗迪贤,等.荷脂细胞胆固醇流出的体系构成与偶联关系[J].南华大学学报:医学版, 2004, 32(3): 347-351.
[16] Yancey PG, Jerome WG. Lysosomal cholesterol derived from mildly oxidized low density lipoprotein is resistant to efflux[J]. J Lipid Res, 2001, 42(3): 317-327.
[17] Dhaliwal BS, Steinbrecher UP. Cholesterol delivered to macrophages by oxidized low density lipoprotein is sequestered in lysosomes and fails to efflux normally[J]. J Lipid Res, 2000, 41(10): 1658-1665.
[18] Gelissen IC, Brown AJ, Mander EL, et al. Sterol efflux is impaired from macrophage foam cells selectively enriched with 7-ketocholesterol[J]. J Biol Chem, 1996, 271(30): 17852-17860.
[19] Young SG, and C. J. Fielding. The ABCs of cholesterol efflux[J]. Nat. Genet, 1999, 22: 316-318.
[20]唐朝克,杨永宗.三磷酸腺苷结合盒转运体A1在THP-1巨噬细胞源性泡沫细胞胆固醇流出中的作用[J].中国病理生理杂志, 2003, 19(8): 1084-1088.
[21]冯翔,凌文华.氧化低密度脂蛋白及溶血卵磷脂对巨噬泡沫细胞胆固醇外流的影响[J].中国病理生理杂志, 2003, 19(9): 1246-1249.
[22]吕湛.胆固醇流出调节蛋白与胆固醇逆向转动和动脉粥样硬化[J].国外医学:生理病理科学与临床分册, 2002, 22(4): 326-328.
[23] Smart EJ, Y.S. Ying, W.C. Donzell, et al. A role for caveolin in transport of cholesterol from endoplasmic reticulum to plasma membrane[J]. J Biol Chem, 1996, 271: 29427-29435.
[24] Uittenbogaard A, Ying Y, Smart EJ. Characterization of a cytosolic heat-shock protein-caveolin chaperone complex. Involvement in cholesterol trafficking[J]. J Biol Chem, 1998, 273(11): 6525-6532.
[25]严鹏科,廖端芳,杨永宗. Caveolin-1表达对血管平滑肌细胞胆固醇逆转运的调节作用[J].中国动脉硬化杂志, 2002, l0(5): 379-383.
[26]王蓉蓉,严鹏科,廖端芳,等.囊泡-囊泡素1在清道夫受体AⅠ转基因小鼠动脉粥样硬化发生中的作用[J].中国动脉粥样硬化杂志, 2002, 10(6): 461-464.
[27] Takahashi N, Hayano T, Suzuki M. Peptidyl-prolyl cis-trans isomerase is the cyclosporinA binding protein cyclophilin[J]. Nature, 1989, 337(6206): 473-475.
[28] Maki K, Ikura T, Hayano T, et al. Effects of proline mutations on the folding of staphylococcal nuclease[J]. Biochemistry, 1999, 38(7): 2213-2223.
[29] Zhao Y, Ke H. Crystal structure implies that cyclophilin predominantly catalyzes the trans to cis isomerization[J]. Biochemistry, 1996, 35(23): 7356-7361.
[30] Jing ZG, Melaragno MG, Liao DF, et al. CyclophilinA is a secreted growth factor induced by oxidative stress[J]. Circ Res, 2000, 87(9): 789-796.
[31] Jin ZG, Lungu AO, Xie L, et al. Cyclophilin A Is a Proinflammatory Cytokine that Activates Endothelial Cells[J]. Arterioscler Thromb Vasc Biol, 2004, 24(7): 1186-1191.
[32] Piotukh K, Gu W, Kofler M, et al. Cyclophilin A binds to linear peptide motifs containing a consensus that is present in many human proteins[J]. J Biol Chem, 2005, 280(25): 23668-23674.
[33]丁振华.低密度脂蛋白的氧化修饰及其理化,生物学性质的改变[J].国外医学:生理病理科学与临床分册, 1994, 14(2): 108-110.
[34]蒋佩,严鹏科,莫中成,等.促进细胞内胆固醇流出抑制单核细胞源性泡沫细胞凋亡[J ].中国病理生理杂志, 2005, 21 (6) : 1041- 1045.
[35]董军,陈文祥,李健斋.高效液相色谱测定微量胆固醇氧化产物[J].生物化学与生物物理进展, 1996, 23: 179-182.
[36] Takahashi N, Hayano T, Suzuki M. Peptidyl-prolyl cis-trans isomerase is the cyclosporinA binding protein cyclophilin[J]. Nature, 1989, 337(6206): 473-475.
[37] Maki K, Ikura T, Hayano T, et al. Effects of proline mutations on the folding of staphylococcal nuclease[J]. Biochemistry, 1999, 38(7): 2213-2223.
[38] Annette Uittenbogaard, Eric J Smart. Palmitoylation of Caveolin-1 is required forcholesterol binding, chaperone complex formation, and rapid transport of cholesterol to caveolae[J]. J Biol Chem, 2000, 75(33): 25595-25599.
[39] Kallen J, Walkinshaw MD. The X-ray structure of a tetrapeptide bound to the active site of human Cyclophilin A[J]. FEBS Lett, 1992, 300(3): 286-290.
[40] Zhao Y, Ke H. Crystal structure implies that cyclophilin predominantly catalyzes the trans to cis isomerization[J]. Biochemistry, 1996, 35(23): 7356-7361.
[41]涂剑,严鹏科,王北冰,等.亲环素A在巨噬细胞荷脂过程中的影响[J ].中国动脉硬化杂志, 2005, 13 (5): 541-544.
[1] Levitsky K, Boersma MD, Ciolli CJ, et al. Exo-Mechanism Proximity- Accelerated Alkylations: Investigations of Linkers, Electrophiles and Surface Mutations in Engineered Cyclophilin-Cyclosporin Systems[J]. Chembiochem, 2005, 6(5): 890-899.
[2] Uittenbogaard A, Everson WV, Matveev SV, et al. Cholesteryl ester is transported from caveolae to internal membranes as part of a caveolin-annexin II lipid-proteincomplex[J]. J Biol Chem, 2002, 277(7):4925-4931.
[3] Piotukh K, Gu W, Kofler M, et al. Cyclophilin A binds to linear peptide motifs containing a consensus that is present in many human proteins[J]. J Biol Chem, 2005, 280(25): 23668-23674.
[4] Jin ZG, Lungu AO, Xie L, et al. Cyclophilin A Is a Proinflammatory Cytokine that Activates Endothelial Cells[J]. Arterioscler Thromb Vasc Biol, 2004, 24(7):1186- 1191.
[5] Jing ZG, Melaragno MG, Liao DF, et al. Cyclophilin A is a secreted growth factor induced by oxidative stress[J]. Circ Res, 2000, 87(9): 789-796.
[6] Bennett MR , Even GI , Schwartz SM, et al. Apoptosis of hunan vascular smooth musde cells derived from noumal vessels and coronary atherosclerotic plaques [ J ] . J Clin Invest, 1995, 95 (5) : 2266– 2274.
[7]唐朝克,杨永宗.三磷酸腺苷结合盒转运体A1在THP-1巨噬细胞源性泡沫细胞胆固醇流出中的作用.中国病理生理杂志, 2003, 19(8): 1084-1088.
[8]冯翔,凌文华.氧化低密度脂蛋白及溶血卵磷脂对巨噬泡沫细胞胆固醇外流的影响.中国病理生理杂志, 2003, 19(9): 1246-1249.
[9]吕湛.胆固醇流出调节蛋白与胆固醇逆向转动和动脉粥样硬化.国外医学:生理病理科学与临床分册, 2002, 22(4): 326-328.
[10]廖端芳,庹勤慧,罗迪贤,等.荷脂细胞胆固醇流出的体系构成与偶联关系.南华大学学报·医学版, 2004, 32(3): 347-351.
[11]Andreea O, Lungu, Zheng - Gen Jin, et al. Cyclosporin A inhibits flow - mediated activation of endothelial nitric-oxide synthase by altering cholesterol content in caveolae[J]. J Biol Chem, 2004, 279: 48794-48800.
[12]Awasthi V, Mandal SK, Papanna V, et al. Modulation of Tissue Factor-Factor VIIa Signaling by Lipid Rafts and Caveolae[J]. Arterioscler Thromb Vasc Biol, 2007, 27(6): 1447-1455.
[13]Covey SD, Brunet RH, Gandhi SG, et al. Cholesterol depletion inhibits fatty acid uptake without affecting CD36 or caveolin-1 distribution in adipocytes[J]. Biochem Biophys Res Commun, 2007, 355(1):67-71.
[14]Theriault Y, Logan TM, Meadows R, et al. Solution structure of the cyclosphorin A/cyclophilin complex by NMR [J]. Nature, 1993, 361: 88-91.
[15]Muruyama T, Frutani M. Archaeal peptidyl prolyl cis-trans isomerases(PPIases)[J].Front Biosci, 2000, 5: 821-836.
[16]Luban J, Bossolt KL, Franke EK, et al. Human immunodeficiency virus type 1 Gag protein binds to cyclophilinsA and B[J]. Cell, 1993, 73(6):1067-1078.
[17]Scholz I, Arvidson B, Huseby D, et al. Virus particle core defects caused by mutations in the human immunodeficiency virus capsid N-terminal domain[J]. J Virol, 2005, 79(3): 1470-1479.
[18]Scholz C, Schaarschmidt P, Engel AM, et al. Functional solubilization of aggregation-prone HIV envelope proteins by covalent fusion with chaperone modules[J]. J Mol Biol, 2005, 345(5):1229-1241.
[19]Chatellard-Gruaz D, Saurat JH, Siegenthaler G. Differential expression of cyclophilin isoforms during keratinocyte differentiation[J]. Biochem J, 1994, 303 (3):863-867.
[20]Watanabe S, Tsuruoka S, Vijayakumar S, et al. Cyclosporin A produces distal renal tubular acidosis by blocking peptidyl prolyl cis-trans isomerase activity of cyclophilin[J]. Am J Physiol Renal Physiol, 2005, 288(1): F40-47.
[21]涂剑,严鹏科,王北冰,等.亲环素A在巨噬细胞荷脂过程中的影响[J].中国动脉硬化杂志, 2005, 13(5): 541-544.
[22]廖端芳,庹勤慧,罗迪贤,等.荷脂细胞胆固醇流出的体系构成与偶联关系.南华大学学报·医学版, 2004, 32(3): 347-351.
[23]廖端芳,严奉祥. Caveolin-1、CyPA与动脉粥样硬化的关系.中国病理生理杂志, 2002, 18(13): 1663-1665.
[24]Zhang YM, Wang KQ, Zhou GM, et al. Endothelin– 1 promoted proliferation of vascular smooth muscle cell through pathway of extracellular signal - regulated kinase and cyclin D1 [J] . Acta Pharmacol Sin, 2003, 24: 563 -568.
[25]Yang H, Li M, Chai H, et al. Effects of cyclophilin A on cell proliferation and gene expressions in human vascular smooth muscular cells and endothelial cells [J]. J Surg Res, 2005, 123(2) :312 - 319.
[26]周四桂,雷小勇,廖端芳.缺氧预适应对缺氧复氧诱导内皮细胞-中性粒细胞黏附的影响[J].中国危重病急救医学, 2003, 15(3): 159-163.
[27]涂光辉,王北冰,严鹏科,等.亲环素A在巨噬细胞荷脂过程中的表达及普罗布考干预的影响[J].中国动脉硬化杂志, 2006, 14(4): 293-296.
[28]Geng YJ, Libby K. Evidence for apoptosis in advanced human[J]. A m J Pathol, 1995, 147 (2) : 251– 266.
[29]Han DK, Haudenschild CC , Hong MK, et al. Evidence for apoptosis in human atherogenesis and in a rat vascular injury model [J]. A m J Pathol, 1995, 147 (2): 267– 277.
[30]Li Z, Zhao X, Bai S, et al. Proteomic identification of cyclophilin A as a potential prognostic factor and therapeutic target in endometrial carcinoma. Mol Cell Proteomics, 2008, 7(10): 1810-1823.