Objective To explore the possible mechanism of Wuzi Yanzong Pills（五子衍宗丸） on apoptotic pathway of neural tube defects（NTDs） cell model.Methods PC12 cells were randomly divided into a blank group

a model group and Wuzi Yanzong Pills low

medium

high dose group.All groups but the blank group used all-trans retinoic acid（atRA） to intervene PC12 cells to construct NTDs model.The low

medium and high doses of Wuzi Yanzong Pills were given RPMI-1640 medium containing Wuzi Yanzong Pills and the final concentration was 125

250

500 μg/m L

respectively for pre-treatment.The model group and the blank group were added with RPMI-1640 medium.The viability and apoptosis rate of PC12 cells in each group were determined

and the total RNA sequencing of the blank group

the model group and the optimal dose of the Wuzi Yanzong Pills group that determined by cell viability and apoptosis rate was extracted.The differentially expressed genes were screened and the target differentially expressed gene set was obtained by Venn diagram

and performed significant enrichment analysis of GO function and KEGG pathway

and also performed co-expressed network analysis of differential genes related to apoptosis.Results The cell viability of PC12 cells in the model group was significantly decreased

and the apoptosis rate was significantly increased.The cell viability of Wuzi Yanzong Pills groups were increased

apoptosis rate decreased

and the best result appeared in the Wuzi Yanzong Pills low dose group（P<0.05 or P<0.01）.Target differentially expressed gene set GO functional analysis was significantly enriched into endoplasmic reticulum stress-mediated apoptosis-associated and cell proliferation-related GO entries

and the KEGG pathway analysis significantly enriched the cell growth and death-related signaling pathways.Compared with the model group

the endoplasmic reticulum stress-related genes Ern1

Casp12 and Creb3l1 were down-regulated in the Wuzi Yanzong Pills low-dose group

and the anti-apoptotic gene Birc5 was up-regulated

which was in the opposite direction to the genes in the model group compared with the blank group.Conclusion Wuzi Yanzong Pills can inhibit apoptosis

protect neuron-like cells

and its mechanism might be related to the regulation of gene expression related to endoplasmic reticulum stress

and biological processes or signal pathways such as chromosome segregation

cell division

cell cycle and other cell proliferation

growth and death.