电离辐射诱导椎间盘纤维环细胞衰老的研究

The investigation of ionizing radiation-induced senescence in annulus fibrosus cells of intervertebral discs.

摘要:

摘要: ［目的］探讨电离辐射对大鼠纤维环细胞衰老及衰老相关分泌表型表达的影响。［方法］选取6只清洁级成熟SD大鼠，取其腰尾段椎间盘纤维环细胞做体外细胞培养，将同等条件下培养的纤维环细胞随机分为两组，分别接受剂量为0Gy、15Gy的X线电离辐射，随后通过细胞形态学观察、β-半乳糖苷酶（SA-β-Gal）染色、细胞增殖检测（CCK-8)、IF检测（Immunofluorescence，IF）及PCR检测等实验方法，研究不同组别纤维环细胞的衰老情况以及衰老相关分泌表型的表达。［结果］经15Gy电离辐射处理后，纤维环细胞在形态学上表现出典型的衰老特征，SA-β-Gal测定显示纤维环细胞阳性染色百分比为77%。CCK-8实验显示纤维环细胞9天内无增殖表现。IF及PCR检测均显示15Gy组衰老基因P16、P21表达增加，衰老相关分泌表型包括胶原蛋白（collagen，Col）-1、Col-2，白细胞介素（interleukin，IL）-6、IL-8，基质金属蛋白酶（matrix metalloproteinase, MMP）-1、MMP-3的表达较未辐射组明显增强，差异均有统计学意义（P<0.05）。［结论］电离辐射能诱导大鼠椎间盘纤维环细胞衰老，其衰老相关分泌表型的表达明显增加。

Abstract:

Abstract: [Objective] To investigate the effects of ionizing radiation on senescence and senescence related secretion phenotypic expression of rat annulus fibrosus cells. [Methods] Six clean-grade mature SD rats were selected. The lumbar intervertebral disc annulus fibrosus cells from these rats were cultured in vitro under identical conditions. Subsequently, the cultured annulus fibrosus cells were randomly divided into two groups: one group received an ionizing X-ray radiation dose of 0Gy, while the other group received a dose of 15Gy. Morphological observation, SA-β-Gal staining, CCK-8, IF and PCR detection were performed to study the senescence levels and expression of senescence-related secretory phenotypes in different groups. [Results] After exposure to 15Gy ionizing radiation, the annulus fibrosus cells exhibited typical morphological characteristics associated with aging. The SA-β-Gal assay revealed a positive staining rate of 77% in annulus fibrosus cells. Furthermore, the CCK-8 assay demonstrated no proliferation of annulus fibrosus cells within a span of 9 days. In addition, the expression levels of aging-related genes P16 and P21 were upregulated in the 15Gy group. Moreover, there was a significant increase in the expression levels of aging-related secretion phenotypes such as collagen Col-1 and Col-2, inflammatory factors IL-6 and IL-8, as well as matrix metalloproteinases MMP-1 and MMP-3 when compared to the control group (P<0.05). [Conclusion] The exposure to ionizing radiation can induce senescence in rat intervertebral disc annulus cells, leading to a significant increase in the expression of SASP.