姜黄素对慢性阻塞性肺疾病患者外周血内皮祖细胞血管生成能力的影响
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摘要
目的探讨姜黄素防治慢性阻塞性肺疾病(COPD)的可行性及可能作用机制。方法 30例COPD患者采用密度梯度离心法从外周血获取单个核细胞,将其接种在人纤维连接蛋白包被的培养板培养7天后,通过荧光染色鉴定为内皮祖细胞(EPCs)。EPCs分成对照组和姜黄素高、中、低剂量组,每组6个样本。对照组用不含胎牛血清的EGM-2培养基培养24 h后,加含20%胎牛血清的EGM-2培养液;姜黄素低、中、高剂量组用不含胎牛血清的EGM-2培养液培养24 h后,再分别用5、10、15μmol/L姜黄素的条件培养液培养。各组培养24 h后检测EPCs增殖能力和血管生成能力。再比较效果最好的姜黄素剂量组和对照组分别培养0 h、12 h、24 h、48 h时EPCs增殖能力和血管生成能力。结果培养24 h时,姜黄素中、高剂量组细胞数量和增殖能力均明显高于对照组,姜黄素各剂量组血管生成能力高于对照组(P <0. 05或P <0. 01),并且随着姜黄素剂量的增加,细胞增殖能力和血管生成能力也增强(P <0. 05或P <0. 01),故选择姜黄素高剂量组与对照组在不同时间进行比较。与对照组同时间比较,姜黄素高剂量组在12 h、24 h、48 h时细胞增殖能力和血管生成能力均升高(P <0. 05或P <0. 01)。结论姜黄素可促进COPD患者EPCs的增殖能力和血管生成能力,且姜黄素剂量为15μmol/L效果最好。
Objective To investigate the feasibility and possible mechanism of curcumin in the prevention and treatment of chronic obstructive pulmonary disease(COPD). Methods A total of 30 patients with COPD were enrolled by collecting peripheral blood using density gradient centrifugation. They were inoculated in human fibronectincoated plates for 7 days and identified as endothelial progenitor cells(EPCs) by fluorescent staining. EPCs were divided into control group and curcumin high,medium and low doses groups,with 6 samples in each group. The control group was cultured in EGM-2 medium without fetal bovine serum for 24 hours,then EGM-2 medium containing 20%fetal bovine serum was added; the curcumin low,medium and high doses groups were treated with fetal bovine serum free EGM-2 culture fluid,and then were cultured with a conditioned medium of 5,10,and 15 μmol/L of curcumin respectively for 24 hours. The proliferation ability and angiogenic ability of EPCs were detected after 24 hours of culture. The proliferation ability and angiogenic ability of EPCs at 0 h,12 h,24 h,and 48 h culture between the curcumin group with best effect dose and the control group were compared. Results After 24 hours of culture,the number and proliferation in the curcumin medium and high dose groups was significantly higher than that in the control group. The angiogenic ability of the curcumin groups was higher than that in the control group(P < 0. 05 or P <0. 01). With the increase of curcumin dose,cell proliferation and angiogenesis ability also increased(P < 0. 05 or P < 0. 01),so the high-dose curcumin group and the control group were compared at different times. Compared with the control group at the same time,the cell proliferation and angiogenesis ability of the curcumin high-dose group were increased at 12 h,24 h,and 48 h(P < 0. 05 or P < 0. 01). Conclusion Curcumin could promote the proliferation and angiogenesis of EPCs in patients with COPD,and the curcumin dose at 15 μmol/L produced best effect.
Objective To investigate the feasibility and possible mechanism of curcumin in the prevention and treatment of chronic obstructive pulmonary disease(COPD). Methods A total of 30 patients with COPD were enrolled by collecting peripheral blood using density gradient centrifugation. They were inoculated in human fibronectincoated plates for 7 days and identified as endothelial progenitor cells(EPCs) by fluorescent staining. EPCs were divided into control group and curcumin high,medium and low doses groups,with 6 samples in each group. The control group was cultured in EGM-2 medium without fetal bovine serum for 24 hours,then EGM-2 medium containing 20%fetal bovine serum was added; the curcumin low,medium and high doses groups were treated with fetal bovine serum free EGM-2 culture fluid,and then were cultured with a conditioned medium of 5,10,and 15 μmol/L of curcumin respectively for 24 hours. The proliferation ability and angiogenic ability of EPCs were detected after 24 hours of culture. The proliferation ability and angiogenic ability of EPCs at 0 h,12 h,24 h,and 48 h culture between the curcumin group with best effect dose and the control group were compared. Results After 24 hours of culture,the number and proliferation in the curcumin medium and high dose groups was significantly higher than that in the control group. The angiogenic ability of the curcumin groups was higher than that in the control group(P < 0. 05 or P <0. 01). With the increase of curcumin dose,cell proliferation and angiogenesis ability also increased(P < 0. 05 or P < 0. 01),so the high-dose curcumin group and the control group were compared at different times. Compared with the control group at the same time,the cell proliferation and angiogenesis ability of the curcumin high-dose group were increased at 12 h,24 h,and 48 h(P < 0. 05 or P < 0. 01). Conclusion Curcumin could promote the proliferation and angiogenesis of EPCs in patients with COPD,and the curcumin dose at 15 μmol/L produced best effect.
引文
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[2]郭瑛,李燕芹,刘斌,等.血管内皮功能变化与慢性阻塞性肺疾病的关系[J].上海交通大学学报(医学版),2010,30(9):1152-1155.
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[7]KALKA C,MASUDA H,TAKAHASHI T,et al. Transplantation of ex vivo expanded endothelial progenitor cells for therapeutic neovascularization[J]. Proc Natl Acad Sci USA,2000,97(7):3422-3427.
[8]VASA M,FICHTLSCHERER S,AICHER A,et al. Number and migratory activity of circulation endothelial progenitor cells inversely correlate with risk factors for coronary artery disease[J]. Cir Res,2001,89(1):E1-E7.
[9]PEICHEV M,NAIYER AJ,PEREIRA D,et al. Expression of VEGFR-2 and AC133 by circulating human CD34(+)cells identifies a population of functional endothelial precursors[J]. Blood,2000,95(3):952-958.
[10]MUROHARA T,TEPPER O,SILVER M,et al. Determination of bone marrow-derived endothelial progenitor cell significance in angiogenic growth factor-induced neovascularization in vivo[J]. Exp Hematol,2002,30(8):967-972.
[11]SUZUKI T,NISHIDA M,FUTAMI S,et al. Neoendothelialization after peripheral blood stem cell transplantation in humans. A case report of a Tokaimura nuclear accident victim[J]. Cardiovasc Res,2003,58(2):487-492.
[12]LIU X,XIE C. Human endothelial progenitor cells isolated from COPD patients are dysfunctional[J]. Mol Cell Biochem,2012,363(1/2):53-63.
[13]曹宏,刁路明,夏东.姜黄素与顺铂联合对人肺腺癌A549细胞增殖和凋亡作用[J].武汉大学学报,2008,29(3):213-217.
[14]BARNES PJ,AADCOCK IM. NF-Kappa B:a pivotal role in asthma and a new target therapy[J]. Trends Pharmacol Sci,1997,18(2):46-50.
[15]狄建彬,顾振纶,赵笑东,等.姜黄素的抗氧化和抗炎作用研究进展[J].中草药,2010,41(5):188-191.
[16]张立,张淑芳,邹维.姜黄素诱导肿瘤细胞凋亡机制研究进展[J].吉林医学,2010,31(13):185-186.
[17]PASCHALAKI KE,STARKE RD,HU Y,et al. Dysfunction of endothelial progenitor cells from smokers and COPD patients due to increased DNA damage and senescence[J]. Stem Cells,2013,31(12):2813-2826.
[2]郭瑛,李燕芹,刘斌,等.血管内皮功能变化与慢性阻塞性肺疾病的关系[J].上海交通大学学报(医学版),2010,30(9):1152-1155.
[3]FADINI GP,SEHIAVON M,CANTINI M,et al. Circulating progenitor cells are reduced in patients with severe lung disease[J]. Stem Cells,2006,24(7):1806-1813.
[4]PALANGE P,TESTA U,HUERTAS A,et al. Cireulating haemopoietic and endothelial progenitor cells are decreased in COPD[J]. Eur Respir J,2006,27(3):529-541.
[5]吴钟伟,张怀勤,黄伟剑,等.姜黄素对内皮祖细胞功能及内皮型一氧化氮合酶的影响[J].心脏杂志,2008,20(2):136-141.
[6]中华医学会呼吸病学分会慢性阻塞性肺疾病学组.慢性阻塞性肺疾病诊治指南(2013年修订版)[J].中华结核和呼吸杂志,2013,36(4):255-264.
[7]KALKA C,MASUDA H,TAKAHASHI T,et al. Transplantation of ex vivo expanded endothelial progenitor cells for therapeutic neovascularization[J]. Proc Natl Acad Sci USA,2000,97(7):3422-3427.
[8]VASA M,FICHTLSCHERER S,AICHER A,et al. Number and migratory activity of circulation endothelial progenitor cells inversely correlate with risk factors for coronary artery disease[J]. Cir Res,2001,89(1):E1-E7.
[9]PEICHEV M,NAIYER AJ,PEREIRA D,et al. Expression of VEGFR-2 and AC133 by circulating human CD34(+)cells identifies a population of functional endothelial precursors[J]. Blood,2000,95(3):952-958.
[10]MUROHARA T,TEPPER O,SILVER M,et al. Determination of bone marrow-derived endothelial progenitor cell significance in angiogenic growth factor-induced neovascularization in vivo[J]. Exp Hematol,2002,30(8):967-972.
[11]SUZUKI T,NISHIDA M,FUTAMI S,et al. Neoendothelialization after peripheral blood stem cell transplantation in humans. A case report of a Tokaimura nuclear accident victim[J]. Cardiovasc Res,2003,58(2):487-492.
[12]LIU X,XIE C. Human endothelial progenitor cells isolated from COPD patients are dysfunctional[J]. Mol Cell Biochem,2012,363(1/2):53-63.
[13]曹宏,刁路明,夏东.姜黄素与顺铂联合对人肺腺癌A549细胞增殖和凋亡作用[J].武汉大学学报,2008,29(3):213-217.
[14]BARNES PJ,AADCOCK IM. NF-Kappa B:a pivotal role in asthma and a new target therapy[J]. Trends Pharmacol Sci,1997,18(2):46-50.
[15]狄建彬,顾振纶,赵笑东,等.姜黄素的抗氧化和抗炎作用研究进展[J].中草药,2010,41(5):188-191.
[16]张立,张淑芳,邹维.姜黄素诱导肿瘤细胞凋亡机制研究进展[J].吉林医学,2010,31(13):185-186.
[17]PASCHALAKI KE,STARKE RD,HU Y,et al. Dysfunction of endothelial progenitor cells from smokers and COPD patients due to increased DNA damage and senescence[J]. Stem Cells,2013,31(12):2813-2826.