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脂多糖抑制人退变髓核细胞 NF-κB 通路炎症与分解代谢

LIPUS inhibits inflammation and catabolism through the NF-κB pathway in human degenerative nucleus pulposus cells.

机构信息

Department of Rehabilitation Medicine, The First Affiliated Hospital of Chongqing Medical University, No. 1 Shiyou Street, Yuzhong District, Chongqing, 400010, China.

Laboratory Research Center, The First Affiliated Hospital of Chongqing Medical University, No. 1 Shiyou Street, Yuzhong District, Chongqing, 400010, China.

出版信息

J Orthop Surg Res. 2021 Oct 18;16(1):619. doi: 10.1186/s13018-021-02739-1.

Abstract

BACKGROUND

Low-intensity pulsed ultrasound (LIPUS) is a safe and noninvasive rehabilitative physical therapy with anti-inflammatory effects. The current study investigated the effect of LIPUS on the inflammation of nucleus pulposus (NP) cells and its underlying mechanism.

METHODS

Human NP cells were acquired from lumbar disc herniation tissue samples and cultured for experiments. Human NP cells were treated with LPS and then exposed to LIPUS (15 mW/cm, 30 mW/cm and 60 mW/cm) for 20 min daily for 3 days to determine the appropriate intensity to inhibit the expression of the inflammatory factors TNF-α and IL-1β. The gene and protein expression of aggrecan, collagen II, MMP-3 and MMP-9 was measured by real-time PCR and western blotting, respectively. The activity of the nuclear factor-kappa B (NF-κB) pathway was examined by western blotting and immunofluorescence. After pretreatment with the NF-κB inhibitor PDTC, the expression of TNF-α, IL-1β, MMP-3 and MMP-9 was measured by real-time PCR.

RESULTS

LIPUS at intensities of 15 mW/cm, 30 mW/cm and 60 mW/cm inhibited LPS-induced NP cell expression of the inflammatory factors TNF-α and IL-1β, especially at 30 mW/cm. LIPUS significantly upregulated the gene and protein expression of aggrecan and collagen II and downregulated the gene and protein expression of MMP-3 and MMP-9 in LPS-induced NP cells. The NF-κB signaling pathway was inhibited by LIPUS through inhibiting the protein expression of p-P65 and the translocation of P65 into the nucleus in LPS-induced NP cells. In addition, LIPUS had similar effects as the NF-κB inhibitor PDTC by inhibiting the NF-κB signaling pathway, inflammation and catabolism in LPS-induced human degenerative nucleus pulposus cells.

CONCLUSION

LIPUS inhibited inflammation and catabolism through the NF-κB pathway in human degenerative nucleus pulposus cells.

摘要

背景

低强度脉冲超声(LIPUS)是一种安全、非侵入性的康复物理疗法,具有抗炎作用。本研究探讨了 LIPUS 对髓核细胞炎症的影响及其机制。

方法

从腰椎间盘突出症组织标本中获取人 NP 细胞进行实验。将人 NP 细胞用 LPS 处理后,每天用 LIPUS(15mW/cm、30mW/cm 和 60mW/cm)照射 20min,连续 3 天,以确定抑制炎症因子 TNF-α和 IL-1β表达的适宜强度。实时 PCR 和 Western blot 分别检测聚集蛋白聚糖、胶原 II、MMP-3 和 MMP-9 的基因和蛋白表达。Western blot 和免疫荧光法检测核因子-κB(NF-κB)通路的活性。用 NF-κB 抑制剂 PDTC 预处理后,通过实时 PCR 检测 TNF-α、IL-1β、MMP-3 和 MMP-9 的表达。

结果

LIPUS 在 15mW/cm、30mW/cm 和 60mW/cm 强度下抑制 LPS 诱导的 NP 细胞炎症因子 TNF-α和 IL-1β的表达,尤其是 30mW/cm 时。LIPUS 显著上调 LPS 诱导的 NP 细胞中聚集蛋白聚糖和胶原 II 的基因和蛋白表达,下调 MMP-3 和 MMP-9 的基因和蛋白表达。LIPUS 通过抑制 LPS 诱导的 NP 细胞中 p-P65 蛋白表达和 P65 向核内易位,抑制 NF-κB 信号通路。此外,LIPUS 通过抑制 NF-κB 信号通路、炎症和 LPS 诱导的人退变髓核细胞的分解代谢,具有与 NF-κB 抑制剂 PDTC 相似的作用。

结论

LIPUS 通过 NF-κB 通路抑制人退变髓核细胞的炎症和分解代谢。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/2b09/8522043/610b48ac22a3/13018_2021_2739_Fig1_HTML.jpg

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