基于Keap1/Nrf2/HO-1信号通路探讨人参提取物对毛囊干细胞氧化应激损伤的影响  

Discussing impact of Ginseng extract on oxidative stress damage in rat hair follicle stem cells based on Keap1/Nrf2/HO-1 signaling pathway

作　　者：李焕 许佳 杨斌[1] 迟慧彦[1] 宋坪 LI Huan;XU Jia;YANG Bin;CHI Huiyan;SONG Ping(Xiyuan Hospital,China Academy of Chinese Medical Sciences,Beijing 100507,China)

机构地区：[1]中国中医科学院西苑医院皮肤科,北京100507 [2]中国中医科学院广安门医院皮肤科

出　　处：《环球中医药》2025年第4期671-676,共6页Global Traditional Chinese Medicine

基　　金：国家自然科学基金(82474521);北京中西医结合学会科研促进工作委员会科研课题(BJZXY-2024-YY0002)。

摘　　要：目的 研究人参提取物对毛囊干细胞(hair follicle stem cells, HFSCs)的调节作用及其机制。方法 采用不同浓度的人参提取物(30、100、300、1 000、3 000、9 000μg/mL)分别作用于HFSCs 24小时后,CCK-8法检测人参提取物作用于HFSCs的最佳安全有效浓度;取对数生长期的HFSCs株,分为对照组和人参组,CCK-8法检测最佳安全有效浓度的人参提取物干预24、48、72小时后的细胞增殖活性;流式细胞术检测干预24小时后HFSCs凋亡情况;活性氧荧光探针(DCFH-DA)法检测干预后细胞内活性氧(reactive oxygen species, ROS)水平;ELISA法检测细胞内丙二醛(malondialdehyde, MDA)和超氧化物歧化酶(superoxide dismutase, SOD)水平;蛋白质免疫印迹法(Western blot, WB)检测Kelch样ECH相关蛋白1(Kelch like ECH-associated protein 1,Keap1)/核因子E2相关因子2(nuclear factor erythroid 2-related factor 2,Nrf2)/血红素氧合酶-1(Heme oxygenase 1,HO-1)蛋白表达情况。结果 与对照组相比,人参提取物干预HFSCs 24小时后,细胞增殖率无明显差异性,干预48、72小时后,细胞增殖率升高(P<0.05),48小时最为显著;与对照组相比,人参提取物干预HFSCs 24小时后,细胞凋亡率显著降低(P<0.05),ROS、MDA水平降低,SOD水平增高(P<0.05);与对照组相比,人参提取物干预组Keap1、Nrf2、HO-1蛋白表达升高(P<0.05),其中以Nrf2蛋白升高最为显著。结论 人参提取物能够促进HFSCs增殖,并降低细胞内氧化应激水平从而抑制细胞凋亡,其作用机制可能与Keap1/Nrf2/HO-1抗氧化通路有关,具有治疗雄激素性脱发潜力。Objective To explore the regulatory effect of ginseng extract on rat hair follicle stem cells and its mechanism.Methods Different concentrations of ginseng extracts(30,100,300,1000,3000,9000μg/mL)were applied to HFSCs cells for 24 hours,and the cell counting Kit-8(CCK-8)method was used to detect the best,safe and effective concentrations of ginseng extract acting on HFSCs cells.HFSCs cell lines in logarithmic growth phase were divided into control group and ginseng group,and the cell proliferation activity of ginseng extract at 24,48 and 72 hours after intervention with the best safe and effective concentration was detected by CCK-8 method.Flow cytometry was used to detect the apoptosis of HFSCs cells after intervention.Reactive oxygen species fluorescent probe(DCFH-DA)was used to detect the level of intracellular reactive oxygen species(ROS)after intervention.The method of ELISA was used to detect the levels of malondialdehyde(MDA)and superoxide dismutase(SOD)in cells.Western blot(WB)was used to detect the expression of Kelch-like ECH-related protein 1(Keap1)/nuclear factor E2-related factor 2(Nrf2)/heme oxygenase-1(HO-1)protein.Results Compared with the control group,the cell proliferation rate of HFSCs cells was increased after ginseng extract intervened for 24,48 and 72 hours(P<0.05),and the most significant was at the 48 hours.Compared with the control group,the apoptosis rate of HFSCs cells in the ginseng group was significantly decreased after ginseng extract was intervened for 24 hours(P<0.05).Compared with the control group,ginseng extract significantly down-regulated the levels of ROS and MDA,and up-regulated the activity of SOD(P<0.05).Conclusion Ginseng extract can promote the proliferation of HFSCs,reduce the level of intracellular oxidative stress and inhibit apoptosis,and its mechanism of action may be related to the Keap1/Nrf2/HO-1 antioxidant pathway,which has the potential to treat androgenetic alopecia.

关 键 词：雄激素性脱发 人参提取物 毛囊干细胞 Keap1/Nrf2/HO-1信号通路 氧化应激 增殖 凋亡 

分 类 号：R285.5[医药卫生—中药学]