Objective To investigate the effect of Notch1 and Notch2 genes on proliferation and apoptosis of glomerular podocytes. Methods (1) Construction of podocyte Notch1 and Notch2 gene silencing and activation models was conducted. Podocytes were divided into NC group, Notch1-short hairpin RNA (shRNA) group (sh1, sh2, sh3 groups), Notch2⁃shRNA group (sh4, sh5, sh6 groups), or silencing negative control group (SC313 group), as well as empty vector group (VP64 group), Notch1⁃small guide RNA (sgRNA) group (sg1, sg2, sg3 groups), Notch2⁃sgRNA group (sg4, sg5, sg6 groups), or activation negative control group (OC276 group). The real⁃time fluorescent quantitative PCR and Western blot were used to detect mRNA and protein expressions of Notch1 and Notch2 in podocytes to validate the cell models, and the sequences with the highest activation/silencing efficiency were selected for subsequent experiments affter performing corresponding transfections. (2) Cell proliferation and apoptosis assays were conducted. Podocytes were divided into NC group, sh2 group (Notch1⁃silenced group), sh4 group (Notch2⁃silenced group), sg3 group (Notch1⁃activated group), sg5 group (Notch2⁃activated group), SC313 group, OC276 group, or VP64 group. After performing corresponding transfections, CCK⁃8 assay and TUNEL assay were performed to measure podocyte proliferation rate and apoptosis index. Results The Notch1 and Notch2 gene⁃silenced and⁃activated glomerular podocyte models were successfully constructed. The results of real⁃time fluorescent quantitative PCR and Western blot revealed that the sh2, sg3, sh4, and sg5 groups had the highest silencing/activation efficiency. Compared with the NC group, the sh2 and sg5 groups interpreted increased optical density values, whereas the sh4 and sg3 groups exhibited decreased optical density values and increased apoptosis index (P<0.05). Conclusion Overexpression of Notch1 gene and silencing of Notch2 gene both inhibit podocyte proliferation and promote apoptosis of podocytes, indicating that these genes play crucial regulatory roles in podocyte proliferation and apoptosis.

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