摘要
目的:探究淫羊藿乙醇提取物(EEBM)干预间充质脂肪干细胞(ADSCs)延缓去势大鼠衰老的机制.方法:将45只3月龄SPF级雌性SD大鼠进行去势处理后,随机分为模型组、ADSCs治疗组和低、中、高质量浓度EEBM干预ADSCs组(1、50、100 mg·L-1),简称AE低、中、高浓度组,每组9只.尾静脉注射相应细胞混悬液200μL后,通过酶联免疫吸附测定法(ELISA)及蛋白免疫印迹法(Western blot)检测细胞周期蛋白依赖性激酶抑制因子(p21)、人体抑癌基因(p53)、白细胞介素-6(IL-6)、白细胞介素-8(IL-8)、超氧化物歧化酶(SOD)、丙二醛(MDA)、B细胞淋巴瘤-2(Bcl-2)、Bcl-2-相关X蛋白(Bax)、胱天蛋白酶-3(Caspase-3)及脂褐素等衰老相关指标.结果:与模型组比较,AE低、中、高浓度组的IL-6含量明显降低(P＜0.05);ADSCs治疗组、AE低、中、高浓度组的脂褐素、MDA及IL-8含量显著降低(P＜0.01),而SOD含量则明显增加(P＜0.05,P＜0.01).与ADSCs治疗组比较,AE低、中、高浓度组的脂褐素、IL-8含量明显减少(P＜0.05,P＜0.01).AE中浓度组的MDA含量明显降低(P＜0.01).与模型组比较,ADSCs治疗组、AE低、中、高浓度组的p21、p53、Bax和Caspase-3蛋白含量明显降低(P＜0.05,P＜0.01),而Bcl-2蛋白含量显著升高(P＜0.01).与ADSCs治疗组比较,AE低、中、高浓度组的p21、p53、Bax和Caspase-3蛋白含量明显降低(P＜0.05,P＜0.01),AE低浓度组的Bcl-2蛋白含量显著升高(P＜0.01).结论:该研究结果显示,EEBM干预的ADSCs或ADSCs可能通过抑制细胞凋亡,减少细胞周期抑制因子和促炎因子,增强抗氧化能力,减少氧化反应,从而延缓去势大鼠衰老,且EEBM干预的ADSCs延缓衰老的效果强于ADSCs,为临床使用EEBM干预ADSCs延缓衰老提供了实验依据.
Abstract
Objective:To explore the mechanism by which the ethanol extract of Epimedium brevicornu(EEBM)intervenes in mesenchymal adipose-derived stem cells(ADSCs)to delay aging in castrated rats.Methods:Forty-five 3-month-old SPF female SD rats were ovariectomized and randomly divided into model group,ADSCs treatment group,and ADSCs groups treated with low,medium,and high concentrations of EEBM(1,50,100 µg·L-1),referred to as the AE low,medium,and high concentration groups,with 9 rats in each group.After tail vein injection of 200 μL of the corresponding stem cell suspension,aging-related indicators including cyclin-dependent kinase inhibitor(p21),tumor suppressor gene(p53),interleukin-6(IL-6),interleukin-8(IL-8),superoxide dismutase(SOD),malondialdehyde(MDA),B-cell lymphoma-2(Bcl-2),Bcl-2-associated X protein(Bax),cysteine-aspartic acid protease-3(Caspase-3),and lipofuscin were measured using enzyme-linked immunosorbent assay(ELISA)and Western blot.Results:Compared with the model group,the IL-6 content in the AE low,medium,and high concentration groups was significantly decreased(P＜0.05).Lipofuscin,MDA,and IL-8 levels in the ADSCs treatment group and AE low,medium,and high concentration groups were significantly reduced(P＜0.01),while SOD content was significantly increased(P＜0.05,P＜0.01).Compared with the ADSCs treatment group,lipofuscin and IL-8 levels in the AE low,medium,and high concentration groups were significantly reduced(P＜0.05,P＜0.01).The MDA content was significantly decreased in the AE medium concentration group(P＜0.01).Compared with the model group,protein levels of p21,p53,Bax,and Caspase-3 in the ADSCs treatment group and AE low,medium,and high concentration groups were significantly reduced(P＜0.05,P＜0.01),while the Bcl-2 protein level was significantly increased(P＜0.01).Compared with the ADSCs treatment group,protein levels of p21,p53,Bax,and Caspase-3 in the AE low,medium,and high concentration groups were significantly reduced(P＜0.05,P＜0.01),and the Bcl-2 protein level in the AE low concentration group was significantly increased(P＜0.01).Conclusion:The results of this experiment show that EEBM-treated ADSCs or ADSCs may delay aging in castrated rats by inhibiting cell apoptosis,reducing cell cycle inhibitors and pro-inflammatory factors,enhancing antioxidant capacity,and reducing oxidative reactions.Moreover,EEBM-treated ADSCs demonstrate stronger anti-aging effects than ADSCs alone.This study provides experimental evidence supporting the clinical use of EEBM to intervene in ADSCs and delay aging.
国家科技期刊平台
NSTL
维普
万方数据