穿心莲内酯通过调控PI3K/Akt炎症通路抑制TBHP诱导髓核细胞凋亡
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武汉市第三医院

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武汉市卫健委医学科研项目


Andrographolide inhibits TBHP-induced apoptosis in nucleus pulposus cells by regulating PI3K/Akt inflammatory pathway
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Wuhan Third Hospital

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    摘要:

    目的:探讨穿心莲内酯(andrographolide,Andro)通过调控PI3K/Akt通路对叔丁基过氧化氢(Tert-butyl hydrogen peroxide,TBHP)诱导髓核细胞凋亡的作用及其机制。方法:CCK-8检测Andro毒性,筛选浓度和最佳处理时间,100 μM TBHP作用24 h构建凋亡模型。将细胞分为正常对照组、TBHP诱导组、TBHP+Andro组、TBHP+Andro+PI3K抑制剂组和TBHP+PI3K抑制剂组。流式细胞术检测细胞凋亡情况,ELISA检测炎性细胞因子TNF-α、IL-1β、IL-6的表达水平。Western blot检测细胞中Bcl-2、Bax、p-Akt和Akt的表达水平。结果:Andro低、中、高剂量为9、18、36 μM,最佳处理时间为24 h。PI3K抑制剂处理后TNF-α、IL-1β、IL-6水平和细胞凋亡率增加(P?0.01),Bcl-2、Bax和p-Akt表达水平无明显变化(P?0.05)。Andro干预后TNF-α、IL-1β、IL-6水平和细胞凋亡率下降(P?0.01),Bcl-2和p-Akt表达水平显著增加,Bax的表达水平显著降低(P?0.01)。结论:Andro能抑制细胞凋亡,降低炎性细胞因子的水平,调控PI3K/Akt炎症通路来抑制TBHP诱导的髓核细胞凋亡。

    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.

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  • 收稿日期:2022-02-16
  • 最后修改日期:2022-06-14
  • 录用日期:2022-09-27
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