基于AMPK/SIRT1/NF-κB通路探讨车前子及其盐炙品延缓小鼠衰老的作用机制OA

To investigate the effects of Plantaginis Semen(PS)and its salt-processed product on D-galactose(D-gal)-induced aging in mice and elucidate the underlying mechanism based on the adenosine monophosphate-activated protein kinase(AMPK)/sirtuin 1(SIRT1)/nuclear factor-kappa B(NF-κB)pathway,and clarify the mechanism of action by which PS and its salt-processed product in delaying aging.A total of 48 mice were randomly divided into the blank control group,model group,low-,medium-,and high-dose PS groups(0.9,1.8,3.6 g·kg-1),and low-,medium-,and high-dose salt-processed PS groups(at the same doses as the PS groups).Except for the blank control group,all groups were subcutaneously injected with D-gal on the nape and back to establish an aging model,which lasted for 60 consecutive days.Starting from the 30th day of modeling,the PS group and the salt-processed PS groups received corresponding drug interventions by gavage for 30 consecutive days,while the blank control group and model group received an equal volume of distilled water using the same method.During the administration period,cognitive function was assessed using the nest-building score and the Morris water maze test.Subsequently,mouse tissue were collected under anesthesia to calculate the organ indices of the kidneys and liver.Hematoxylin-eosin(HE)staining was used to observe the pathological changes of renal tissue.Enzyme-linked immunosorbent assay(ELISA)was employed to detect the activity of superoxide dismutase(SOD)and the content of malondialdehyde(MDA)in renal tissue.Real-time quantitative polymerase chain reaction(RT-qPCR)was used to detect the mRNA expression levels of aging-related genes,including AMPK and SIRT1.Western blot was used to detect the protein expression levels of the aging markers AMPK,SIRT1 and NF-κB.The results showed that compared with the model group,the nesting ability of mice in all dose groups of raw PS and salt-processed PS increased to varying degrees.Notably,the nesting score of the medium-dose salt-processed PS group was significantly higher than that of the medium-dose raw PS group(P<0.05).Compared to the blank control group,the model group had a significantly prolonged escape latency and a significantly decreased frequency of platform crossings(P<0.05).After drug intervention,all treatment groups showed a shortened escape latency and an increased number of platform crossings,indicating improved spatial memory compared to the model group.Compared with the model group,all dose groups of raw and salt-processed PS groups significantly increased the kidney and liver indices of aging mice(P<0.05),with the high-dose salt-processed PS group showing significantly higher kidney and liver indices than the high-dose PS group(P<0.01).Histopathological examination revealed that raw PS and salt-processed PS ameliorated renal tissue morphology,resulting in more intact glomerular structures and regular tubular lumens.Furthermore,both raw PS and salt-processed PS significantly increased the activities of SOD in renal tissue(P<0.05)and decreased the content of MDA(P<0.05).Additionally,they significantly up-regulated the mRNA and protein expression levels of AMPK and SIRT1(P<0.05)while down-regulating the protein expression levels of NF-κB(P<0.05).The regulatory effects on these indices were more pronounced in the salt-processed PS groups(P<0.05).In conclusion,both raw PS and its salt-processed product can significantly delay aging by regulating the AMPK/SIRT1/NF-κB signaling pathway and ameliorating oxidative stress.Furthermore,the anti-aging efficacy is enhanced after processing,with the salt-processed product demonstrating superior effects.