摘要
目的:探讨蛭龙活血通瘀胶囊含药血清对低密度脂蛋白(ox-LDL)诱导的巨噬细胞凋亡及泡沫细胞形成的影响,在细胞层面揭示蛭龙活血通瘀胶囊对动脉粥样硬化的治疗作用。方法:佛波酯(PMA)用于诱导人组织细胞淋巴瘤细胞(U937)分化为巨噬细胞,通过CCK-8筛选蛭龙活血通瘀胶囊含药血清的最适处理浓度。将细胞分为正常对照组、模型组、空白血清对照组、蛭龙活血通瘀胶囊含药血清组、蛭龙活血通瘀胶囊联合阴性对照小干扰RNA(si-NC)组、蛭龙活血通瘀胶囊联合去乙酰化转移酶7小干扰RNA(si-SIRT7)组。油红O染色观察泡沫细胞形成情况;CCK-8检测细胞活力;流式细胞术检测细胞凋亡;Western blot检测GRP78、CHOP、Caspase-12、SIRT7蛋白表达。结果:10%蛭龙活血通瘀胶囊含药血清是最适处理浓度。与对照组比较,模型组细胞胞内脂滴增多,细胞大量聚集,细胞活力显著降低,细胞凋亡率显著升高,GRP78、CHOP、Caspase-12蛋白表达显著升高,SIRT7表达显著降低(P<0.01)。蛭龙活血通瘀胶囊含药血清能抑制泡沫细胞形成,显著增加细胞活力并抑制细胞凋亡,还能显著降低GRP78、CHOP、Caspase-12蛋白表达,显著升高蛋白SIRT7表达(P<0.05或P<0.01)。干扰SIRT7的表达逆转了蛭龙活血通瘀胶囊的作用。结论:蛭龙活血通瘀胶囊能有效改善ox-LDL诱导巨噬细胞凋亡及泡沫细胞形成,其机制可能与SIRT7介导的内质网应激有关。
Objective:To investigate the effect of serum containing Zhilong huoxue tongyu capsules on low-density lipoprotein(ox-LDL)-induced macrophage apoptosis and foam cell formation,so as to reveal the therapeutic effect of Zhilong huoxue tongyu capsules on atherosclerosis at the cellular level.Methods:Phorbol-12-myristate-13-acetate(PMA)was used to induce the differentiation of human histocytic lymphoma cells(U937)into macrophages.The optimal concentration of serum containing Zhilong huoxue tongyu capsules was screened by CCK-8.The macrophages were divided into the normal control group,model group,blank serum control group,drug serum containing Zhilong huoxue tongyu capsules group,Zhilong huoxue tongyu capsules combined with negative control small interfering RNA(si-NC)group,Zhilong huoxue tongyu capsules combined with deacetyltransferase 7 small interfering RNA(si-SIRT7)group.Oil red O staining was used to observe the formation of foam cells;CCK-8,flow cytometry,Western blot were used to detect cell viability,cell apoptosis and the expressions of GRP78,CHOP,Caspase-12 and SIRT7 protions,respectively.Results:10%Zhilong huoxue tongyu capsules drug-containing serum was the most suitable concentration.Compared with the normal control group,the intracellular lipid droplets in model group increased,the cell aggregation was large,the cell viability was significantly decreased,the apoptosis rate was significantly increased,the expressions of GRP78,CHOP and Caspase-12 proteins were significantly increased,while the expression of SIRT7 was significantly decreased(P<0.01).Zhilong huoxue tongyu capsules drug-containing serum could inhibit the formation of foam cells,significantly increase cell viability and inhibit cell apoptosis,significantly reduce the expressions of GRP78,CHOP and Caspase-12 proteins and significantly increase the expression of SIRT7 protein(P<0.05 or P<0.01).The interference with SIRT7 expression reversed the effect of Zhilong huoxue tongyu capsules.Conclusion:Zhilong huoxue tongyu capsules can effectively im
作者
刘孟楠
韩梅
秦旭佩
周宇飞
罗钢
周华
杨思进
LIU Meng-nan;HAN Mei;QIN Xu-pei;ZHOU Yu-fei;LUO Gang;ZHOU Hua;YANG Si-jin(Department of Cardiovascular Medicine,Affiliated Hospital of Traditional Chinese Medicine of Southwest Medical University,Luzhou 646000,China;School of Traditional Chinese Medicine,Macao University of Science and Technology,Macao 999078,China)
出处
《中药材》
CAS
北大核心
2021年第6期1463-1469,共7页
Journal of Chinese Medicinal Materials
基金
泸州市科技局项目(2019-RCM-95)
西南医科大学-西南医科大学附属中医医院联合项目(2018XYLH-005)
四川省科技厅项目(2018YFS0543,2020ZD002)
国家自然科学基金项目(82074378)
