Abstract:OBJECTIVE: To explore the effect of Andrographolide (Andro) on Tert-butyl hydrogen peroxide (TBHP) induced apoptosis in nucleus pulposus cells through the regulation of PI3K/Akt pathway. METHODS: The toxicity of Andro was detected by CCK-8, and the optimal concentration and treatment time were screened. After treatment with 100 μM TBHP for 24 h, the apoptosis model was established. Cells were divided into control group, TBHP group, TBHP+Andro group, TBHP+Andro+PI3K inhibitor group and TBHP+PI3K inhibitor group. Cell apoptosis was detected by flow cytometry. The expression levels of inflammatory cytokines TNF-α, IL-1β and IL-6 were detected by ELISA. The protein expression levels of Bcl-2, Bax, p-Akt and Akt were detected by Western blot. RESULTS: The low, medium and high doses of Andro were 9, 18, 36 μM, and the optimal treatment time was 24 h. The levels of TNF-α, IL-1β, IL-6 levels and apoptosis rate increased after PI3K inhibitor treatment (P?0.01), while the expression levels of Bcl-2, Bax and p-Akt expression levels showed no significant difference (P?0.05). TNF-α, IL-1β, IL-6 levels and apoptosis rates decreased after Andro intervention (P?0.01), Bcl-2 and p-Akt expression levels increased significantly and Bax expression levels decreased significantly (P?0.01). CONCLUSION: Andro can inhibit cell apoptosis, reduce the level of inflammatory cytokines, and regulate the PI3K/Akt inflammatory pathway to inhibit TBHP-induced apoptosis in nucleus pulposus cells.