鹿红方对氧糖剥夺模型心脏干细胞自噬和抗凋亡能力的影响  

The Influence of Luhong Formula on Autophagy and Anti-apoptosis of Cardiac Stem Cells with Oxygen-sugar Deprivation Model

作　　者：瞿惠燕[1] 徐基杰[1] 戎靖枫[1] 杨涛[1] 王英杰[1] 刘茜[1] 周华[1] QU Huiyan;XU Jijie;RONG Jingfeng;YANG Tao;WANG Yingjie;LIU Qian;ZHOU Hua(Shuguang Hospital Affiliated to Shanghai University of Traditional Chinese Medicine,Shanghai 201203,China)

机构地区：[1]上海中医药大学附属曙光医院

出　　处：《中西医结合心脑血管病杂志》2019年第12期1794-1799,共6页Chinese Journal of Integrative Medicine on Cardio-Cerebrovascular Disease

基　　金：国家自然科学青年基金项目(No.81403363);国家自然科学面上项目(No.81673753)

摘　　要：目的探讨鹿红方(LHF)对氧糖剥夺模型心脏干细胞(CSCs)自噬和抗凋亡能力的影响。方法将CSCs随机分为对照组(正常培养)与氧糖剥夺造模组(缺血缺氧)。将造模组分为模型组(不予有效干预措施)、自噬阻断到(3-MA)组(给予自噬阻断剂)、鹿红方200μg/mL组(给予LHF200μg/mL)和LHF200+3-MA组(给予LHF200μg/mL+自噬阻断剂)。各组给予相应干预措施后光镜下观察细胞形态,并采用CCK-8法测定细胞凋亡情况;WesternBlot检测自噬相关蛋白Beclin-1、LC3-Ⅱ的表达;RT-PCR检测自噬相关蛋白Beclin-1、LC3-ⅡmRNA的表达。结果光镜下观察可见:LHF200组较模型组和3-MA组凋亡细胞显著减少,LHF200+3-MA组与LHF200组比较,细胞凋亡增加;CCK-8结果显示:LHF200组与模型组比较,CSCs活细胞OD值显著增高(P=0.006);LHF+3-MA组与LHF200组比较,CSCs活细胞OD值下降(P=0.03);WesternBlot结果显示:LHF200组Beclin-1和LC3-Ⅱ蛋白表达较模型组显著降低(P<0.001),但显著高于3-MA组和LHF200+3-MA组(P<0.001);RT-PCR结果显示:LHF200组Beclin-1和LC3-ⅡmRNA表达较模型组显著降低(P<0.001),但显著高于3-MA组和LHF200+3-MA组(P<0.001)。结论LHF可提高氧糖剥夺模型CSCs的抗凋亡能力,其机制与调节自噬有关。Objective To investigate the influence of Luhong Formula (LHF) on autophagy and anti-apoptosis of cardiac stem cells (CSCs) with oxygen-sugar deprivation model.Methods CSCs were randomly divided into the control group (normal culture condition) and sugar/oxygen deprivation model group which include model group,autophagy antagonist(3-MA)group (autophagy antagonist),LHF200 group,and LHF200+3-MA group.The cell morphology of each group was observed under light microscope after corresponding intervention measures,and apoptosis was determined by CCK-8 method.Western Blot was used to detect autophagy related proteins expression of Beclin 1 and LC3-Ⅱ,RT-PCR was used to detect autophagy related proteins expression of Beclin 1 and LC3-ⅡmRNA.Results Microscope observation showed that the cell apoptosis reduced significantly in LHF200 group than those in model group and 3-MA group,and they increased in LHF200+3-MA group than those in LHF200 group.CCK-8 showed that the CSCs OD value of LHF 200 group was significantly higher ( P =0.006) compared with that in model group. The CSCs OD value of LHF200+3- MA group was lower than that of LHF200 group ( P =0.03).Western Blot showed that compared with model group,the proteins expression of Beclin-1 and LC3-Ⅱ in LHF200 group was lower( P <0.001),but the proteins expression of Beclin-1 and LC3-Ⅱ in LHF200 group was higher than that in 3-MA group and LHF200+3-MA group ( P <0.001).RT-PCR showed that compared with model group,the proteins expression of Beclin-1 and LC3-Ⅱ mRNA in LHF200 group was lower( P <0.001),but the proteins expression of Beclin-1 and LC3-ⅡmRNA in LHF200 group was higher than that in 3-MA group and LHF200+3-MA group( P <0.001).Conclusion Luhong Formula can improve apoptosis resistance of cardiac stem cells under sugar/oxygen deprivation partly via autophagy regulation.

关 键 词：心脏干细胞 自噬 凋亡 鹿红方 

分 类 号：R318.5[医药卫生—生物医学工程] R289.5[医药卫生—基础医学]