姜黄素通过血红素加氧酶-1信号通路促进脂肪源干细胞增殖和旁分泌研究  

Curcumin promotes the proliferation and paracrine of ADSCs via HO-1 Pathway

作　　者：刘剑锋[1] 朱平[1] 马守原[1] 陈海旭[2] 王曙霞[1] 李珊[1] 

机构地区：[1]解放军总医院南楼心内科,北京100853 [2]解放军总医院老年医学研究所,北京100853

出　　处：《中国心血管杂志》2015年第3期199-203,共5页Chinese Journal of Cardiovascular Medicine

基　　金：国家自然科学基金主任基金(8135002);解放军总医院临床扶持基金(2013FC-TSYS-2015)~~

摘　　要：目的探讨姜黄素对脂肪源干细胞(ADSCs)增殖和旁分泌的作用及其机制。方法酶法分离、扩增SD大鼠腹股沟的ADSCs,流式细胞仪检测细胞表型;MTT法检测不同浓度姜黄素对细胞生长曲线的影响,筛选最佳姜黄素处理浓度;姜黄素预处理ADSCs 24 h,加用血红素加氧酶-1(HO-1)抑制剂Sn PP阻断HO-1活性;q PCR和Western blot检测HO-1信号在姜黄素处理ADSCs过程中的变化;MTT法检测HO-1活性对姜黄素促进细胞增殖的影响;PCR和Elisa法分别检测姜黄素对ADSCs分泌VEGF和HGF的影响。结果第3代ADSCs的CD29、CD90阳性,CD31、CD34、CD45阴性。姜黄素促进ADSCs增殖的作用呈时间依赖性,最佳处理浓度为10μmol/L。加用Sn PP后,促增殖作用显著降低(P=0.001)。姜黄素能显著促进ADSCs分泌VEGF和HGF(分别为P=0.001和P=0.007),Sn PP能明显抑制姜黄素的促分泌作用[Cur-ADSCs比Cur-ADSCs+Sn PP:VEGF为(712.2±43.1)ng/L比(442.3±33.2)ng/L,P=0.001;HGF为(58.3±3.7)ng/L比(33.7±3.1)ng/L,P=0.001]。在此过程中,HO-1表达的变化方向和姜黄素促进ADSCs增殖和旁分泌的方向一致。结论HO-1介导了姜黄素促进ADSCs增殖和旁分泌的作用。姜黄素是一种有前景的增强干细胞治疗疗效的候选药物。Objective To explore the effects and mechanism of curcumin on the proliferation and paracrine of adipose-derived stem cells（ ADSCs）. Methods ADSCs were isolated through enzymes from SD rats inguina. The phenotypes of ADSCs were analyzed by flow cytometry. The effects of different concentrations of curcumin on cell growth curve were determined through MTT. Following by pretreatment of curcumin on ADSCs for 24 h,the Sn PP,an inhibitor of heme oxygenase-1（ HO-1）,was added to suppress HO-1 activity. PCR and western blotting was used to assess the changes of HO-1 expression during the curcumin treatment. The effects of HO-1 expression on the curcumin promoting proliferation by MTT. q PCR and Elisa were applied to investigate the influences of curcumin on the secretion of VEGF and HGF. Results After three passages,most of adherent ADSCs expressed CD29 and CD90. But they were negative for CD31,CD34 and CD45. The promoting proliferation of curcumin on ADSCs depended on time. The optimal treatment concentration of curcumin was 10 μmol / L. After added Sn PP,the increased proliferation of ADSCs by curcumn pretreatment decreased significantly（ P = 0. 001）. And the enhancing VEGF and HGF secreted by curcumin pretreated ADSCs（ P = 0. 001 and P = 0. 007,respectively） were reduced by treatment of Sn PP[Cur-ADSCs vs. Cur-ADSCs ＋ Sn PP： VEGF：（ 712. 2 ± 43. 1） ng/L vs.（ 442. 3 ± 33. 2） ng/L,P = 0. 001;HGF：（ 58. 3 ± 3. 7） ng / L vs.（ 33. 7 ± 3. 1） ng / L,P = 0. 001]. Importantly,the change trend of HO-1 was parallel with the promoting proliferation and paracrine of ADSCs by pretreatment of curcumin. Conclusions Curcumin promotes the proliferation and paracrine of ADSCs partly via HO-1 signaling pathway. Curcumin is a promising candidate drug for enhancing the therapeutic effects of stem cells.

关 键 词：姜黄素 血红素加氧酶-1 脂肪干细胞 细胞增殖 旁分泌 

分 类 号：R285[医药卫生—中药学]