Display omitted •DNLA and metformin ameliorated cognitive decline in aged SAMP8 mice.•Age-associated neuron loss and ER damage was improved by DNLA and metformin.•DNLA and metformin decreased ER stress-related PERK signaling pathway.•DNLA decreased calpain 1, GSK-3β and Cdk5 activities and Tau hyperphosphorylation.•These beneficial effects of DNLA were comparable to metformin. The senescence-accelerated mouse prone 8 (SAMP8) mice have many pathological features of Alzheimer’s disease (AD) with aging. We previously reported that Dendrobium nobile Lindl alkaloid (DNLA) effectively improved cognitive deficits in multiple Alzheimer’s disease (AD) models. This study further used SAMP8 mice to study the anti-aging effects of DNLA, focusing on endoplasmic reticulum (ER) stress. DNLA and metformin were orally administered to SAMP8 mice starting at 4-month of age for 6 months. Behavioral tests were performed in 10-month-old SAMP8 mice and age-matched SAMR1 control mice. At the end of experiment, neuron damage was evaluated by histology and transmission electron microscopy. ER stress-related proteins were analyzed with Western-blot. DNLA improved learning and memory impairments, reduced the loss of neurons and Nissl bodies in the hippocampus and cortex. DNLA ameliorated ER dilation and swelling in the hippocampal neurons. DNLA down-regulated the protein kinase RNA-like endoplasmic reticulum kinase (PERK) signaling pathway, decreased calpain 1, GSK-3β and Cdk5 activities and the Tau hyper-phosphorylation. The effects of DNLA were comparable to metformin. In summary, DNLA was effective in improving cognitive deficits in aged SAMP8 mice, possibly via suppression of ER stress-related PERK signaling pathway, sequential inhibition of calpain 1, GSK-3β and Cdk5 activities, and eventually reducing the hyper-phosphorylation of Tau.