摘要
目的:基于核因子E2相关因子2(Nrf2)/Kelch样环氧氯丙烷相关蛋白-1(Keap1)信号通路探讨人参肽对D-半乳糖(D-gal)联合AlCl_(3)衰老模型小鼠的影响及分子机制。方法:将50只KM小鼠随机分为对照组、模型组、阳性药组、人参肽低剂量组、人参肽高剂量组,每组10只,阳性药组给予参茸片286.3mg·kg^(-1)·d^(-1);人参肽低、高剂量组分别给予人参肽17.5mg·kg^(-1)·d^(-1)、35mg·kg^(-1)·d^(-1),预给药7d后,除对照组外,其余各组小鼠均给予500mg·kg^(-1)·d^(-1)D-gal皮下注射与70mg·kg^(-1)·d^(-1)-AlCl_(3)灌胃30d联合给药制备衰老模型,应用PCR检测各组小鼠鼠尾组织早老素-1(PS1)的基因表达水平验证模型是否制备成功;通过爬杆实验、负重游泳实验测试小鼠协调能力、抗疲劳能力;应用生化法检测小鼠肝组织中超氧化物歧化酶(SOD)、过氧化氢酶(CAT)活性,丙二醛(MDA)含量;HE染色观察小鼠脑海马组织病理形态,计算小鼠脾脏系数,自然杀伤细胞(NK)实验检测小鼠免疫能力;WesternBlot法检测小鼠脑组织中Nrf2、Keapl、氧化蛋白水解酶(AARE)蛋白表达水平。结果:衰老小鼠PS1的基因表达显著升高(P<0.01),说明模型制备成功,且各药物组小鼠PS1的表达显著降低(P<0.05)。与对照组比较,模型组小鼠爬杆时间显著延长(P<0.01);负重游泳时间显著缩短(P<0.01);肝组织中SOD、CAT活性显著降低(P<0.05,P<0.01),MDA含量显著升高(P<0.01);脾脏指数及NK杀伤率显著降低(P<0.05);脑组织中Nrf2、AARE蛋白表达水平显著下调(P<0.01),Keap1蛋白表达水平显著上调(P<0.01)。与模型组比较,各药物组小鼠爬杆时间显著缩短(P<0.01);负重游泳时间显著延长(P<0.01);肝组织中SOD、CAT活性显著升高(P<0.05,P<0.01),MDA含量显著降低(P<0.01);脾脏指数及NK杀伤率显著升高(P<0.05,P<0.01),Nrf2、AARE蛋白表达水平显著上调(P<0.01),Keapl蛋白表达水平显著下调(P<0.05,P<0.01)。结论:人参肽能改善D-gal联合AlCl_(3)诱导衰老小鼠的协调性、增强抗疲劳和免疫能力,其分子机制可能与激活Nrf2/Keap1信号通路抗氧化相关。
Objective:To investigate the effect and molecular mechanism of ginseng peptide(RST)on D-galactose(D-gal)combined with AlCl_(3)aging model mice based on Nrf2/Keap1 signaling pathway.Methods:Fifty KM mice were randomly divided into five groups,control group,model group,positive group(Ginseng Antler Tablets 286.3 mg·kg^(-1)·d^(-1)),RST lowdose group(RSTL,17.5 mg·kg^(-1)·d^(-1))and RST high-dose group(RSTH,35 mg·kg^(-1)·d^(-1)).After 7 days of administration,mice in four groups except the control group were given subcutaneous injection of D-galactose at the dose of 500 mg·kg^(-1)·d^(-1)and gavage of aluminum trichloride at the dose of 70 mg·kg^(-1)·d^(-1)for 30 days to establish the aging model.The gene expression of PS1 of mice in each group was detected by PCR to verify the success of the aging model.The coordination and fatigue resistance were tested through pole climbing and weight-bearing swimming experiments;the activity of SOD,CAT and the content of MDA in liver tissue of mice were detected by biochemical method;HE staining was used to observe the pathological morphology of brain tissue in each group.The spleen coefficient of mice and NK killing cell test were calculated to detect the immune ability of aging mice.The expression level of Nrf2,Keapl,AARE protein in the brain tissue of aging mice in each group was detected by Western Blot.Results:The gene expression of PS1 in aging mice increased significantly(P<0.01),which indicated that the model was successfully prepared,and the expression of PS1 in mice in each drug group decreased significantly(P<0.05).Compared with the control group,the climbing time of mice in the model group was significantly prolonged(P<0.01).The swimming time with load was significantly shortened(P<0.01).The activities of SOD and CAT in liver tissue decreased significantly(P<0.05,P<0.01),and the content of MDA increased significantly(P<0.01).Spleen index and NK killing rate decreased significantly(P<0.05).The expression levels of Nrf2 and AARE protein in brain tissue were significantly decreased(P<0.01),while the expression level of Keapl protein was significantly increased(P<0.01).Compared with the model group,the climbing time of mice in each drug group was significantly shortened(P<0.01).The swimming time with load was significantly prolonged(P<0.01).The activities of SOD and CAT in liver tissue increased significantly(P<0.05,P<0.01),and the content of MDA decreased significantly(P<0.01).Spleen index and NK killing rate increased significantly(P<0.05,P<0.01),the expression levels of Nrf2 and AARE protein increased significantly(P<0.01),and the expression level of Keapl protein decreased significantly(P<0.05,P<0.01).Conclusion:RST can improve the coordination and anti-fatigue of aging mice induced by D-gal combined with AlCl_(3),and its molecular mechanism may be related to the antioxidant activation of Nrf2/Keapl signaling pathway.
作者
李祯
刘雅鑫
刘健
林嘉楠
曹占鸿
房星宇
白浩楠
安昱
杨擎
李娜
LI Zhen;LIU Yaxin;LIU Jian;LIN Jianan;CAO Zhanhong;FANG Xingyu;BAI Haonan;AN Yu;YANG Qing;LI Na(Laboratory of Molecular Pharmacology,Jilin Ginseng Academic,Changchun University of Chinese Medicine,Changchun 130117,China)
出处
《中华中医药杂志》
CAS
CSCD
北大核心
2023年第8期3594-3599,共6页
China Journal of Traditional Chinese Medicine and Pharmacy
基金
吉林省教育厅科学技术研究项目(No.JJKH20220884KJ)
吉林省科技厅项目(No.20200708056YY)。
