[Influences of aged male parents on learning ability of offspring in SD rats and intervention effect of Wuzi Yanzong Pills based on miR-34a-5p/SIRT1 signaling pathway].

Aged male parents (15-month-old rats) showed significantly reduced fertility parameters compared to young controls.

• The model group exhibited reductions in fertility rate, litter size, and sperm motility and count compared with the control group (P<0.01)
• Impaired testicular spermatogenesis was observed by hematoxylin-eosin staining in the model group
• Increased senescence cells in testicular and epididymal tissue were detected by β-galactosidase staining in the model group
• Elevated miR-34a-5p expression in sperms was observed in the aged model group
• Control group consisted of 8 SD male rats of 3 months old; model group consisted of 8 SD male rats of 15 months old

Offspring of aged male parents showed impaired learning and memory performance, reduced hippocampal neurons, and altered miR-34a-5p/SIRT1/BDNF signaling at 30 days of age.

• The 30-day-old offspring showed impaired learning and memory performance in the Morris water maze (P<0.01)
• Reduced hippocampal neurons were detected by Nissl staining in offspring of aged parents (P<0.01)
• Up-regulated miR-34a-5p expression was found in hippocampal tissue of offspring (P<0.01)
• Down-regulated protein levels of SIRT1 and BDNF in the hippocampus were observed (P<0.01)
• Offspring of aged parents also showed slow growth compared to controls

High-dose Wuzi Yanzong Pills (8 g·kg⁻¹) improved fertility parameters and testicular spermatogenesis in aged male rats compared to the model group.

• High-dose Wuzi Yanzong Pills improved fertilization rate, litter size, sperm motility, sperm count, and testicular spermatogenesis compared with the model group (P<0.05)
• Vitamin C (100 mg·kg⁻¹) also improved the same parameters (P<0.05)
• Compared with the vitamin C group, the high-dose Wuzi Yanzong Pills group showed increases in fertilization rate and litter size (P<0.05)
• Treatment was administered for 12 weeks prior to co-caging with 3-month-old female mice for 5 days

High-dose Wuzi Yanzong Pills improved learning, memory, and hippocampal outcomes in offspring of aged male parents.

• The 30-day-old offspring in the high-dose Wuzi Yanzong Pills group showed enhanced learning and memory capabilities (P<0.05)
• Increased hippocampal neurons and elevated BDNF protein level in the hippocampus were observed in offspring (P<0.05)
• Offspring in the high-dose group showed accelerated growth and development compared to the model group (P<0.05)
• Compared with the vitamin C group, the high-dose Wuzi Yanzong Pills group exhibited accelerated offspring growth (P<0.05) and improved learning and memory abilities (P<0.05)

Wuzi Yanzong Pills regulate miR-34a-5p expression to delay spermatogenic cell senescence, with effects transmitted intergenerationally to improve offspring cognitive function.

• Wuzi Yanzong Pills regulate miR-34a-5p expression to delay spermatogenic cell senescence in the testicular tissue
• The improvement of offspring cognitive function occurs via miR-34a-5p mediated intergenerational transmission
• The miR-34a-5p/SIRT1 signaling pathway was identified as the mechanistic basis for both testicular aging and offspring cognitive effects
• miR-34a-5p expression was quantified in both sperm (parental) and hippocampal tissue (offspring) by qPCR, linking paternal sperm epigenetics to offspring brain function

Low-dose Wuzi Yanzong Pills (2 g·kg⁻¹) showed less pronounced effects compared to high-dose treatment.

• Four treatment groups were tested: aged model, model+high-dose (8 g·kg⁻¹) Wuzi Yanzong Pills, model+low-dose (2 g·kg⁻¹) Wuzi Yanzong Pills, and model+vitamin C (100 mg·kg⁻¹), each with n=8
• The abstract specifically highlights significant improvements for high-dose Wuzi Yanzong Pills and vitamin C groups, without reporting equivalent significant effects for the low-dose group
• The study compared high-dose Wuzi Yanzong Pills directly against vitamin C as an active comparator, with high-dose showing superiority in offspring growth, fertilization rate, litter size, and learning/memory (P<0.05)