Zhongguo gu shang = China journal of orthopaedics and traumatology · 2026
[Regulation of the IL-6 and IL-16 pathway in spinal cord tissue by plasma exosome-mediated miR-125b-5p in a rat model of spinal cord injury].
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Yin Na, Wu Xiaolong, Xie Yan, Zhou Yingjie, Zhuo Hanjie, Wang Qingfeng, Huang Yong
Аннотация
OBJECTIVE: To investigate a novel regulatory pathway mediated by exosomal non-coding RNA in rats with spinal cord injury (SCI).
METHODS: (1) A rat model of spinal cord injury (SCI) was established via T9-T10 spinal cord hemisection method. The success of model establishment was evaluated using the Basso-Beattie-Bresnahan(BBB) scoring scale. Spinal cord tissues were collected from successfully modeled rats and normal control rats, respectively. The levels of inflammatory factors-interleukin-6 (IL-6), interleukin-16 (IL-16), tumor necrosis factor-α (TNF-α), and nuclear transcription factor-κB (NF-κB)-were measured by enzyme-linked immunosorbent assay (ELISA). (2) miRNAs capable of regulating inflammatory factors were identified through literature review and bioinformatics analysis. The regulatory relationship between miR-125b-5p and IL-6/IL-16 was verified using a dual-luciferase reporter gene assay. (3) Spinal cord neuronal cells were cultured in vitro and identified by immunofluorescence. The effects of exosomal miR-125b-5p on the expression of IL-6 and IL-16 in both SCI rat neuronal cells and spinal cord tissues were comparatively analyzed.
RESULTS: Compared with normal control group[(29.81±7.29), (35.88±5.73), (71.67±8.75), and (1 285.01±86.40) pg·mL-1, respectively], the levels of IL-6, IL-16, TNF-α and NF-κB in spinal cord tissue of model group(38.36±5.73), (46.82±7.20), (198.33±20.32), (1 907.47±120.95) pg·mL-1, respectively] increased significantly (all P<0.05). Bioinformatics search results showed that miR-125b-5p could regulate IL-6 and IL-16 simultaneously, and the results of dual luciferase reporter gene experiments confirmed this regulatory relationship directly. Cell and animal experiments have confirmed that exosomal miR-125b-5p could down regulate expression of IL-6 and IL-16.
CONCLUSION: The regulatory pathway regulating IL-6/IL-16 in spinal cord tissue through exosomal miR-125b-5p was identified in SCI rats.
METHODS: (1) A rat model of spinal cord injury (SCI) was established via T9-T10 spinal cord hemisection method. The success of model establishment was evaluated using the Basso-Beattie-Bresnahan(BBB) scoring scale. Spinal cord tissues were collected from successfully modeled rats and normal control rats, respectively. The levels of inflammatory factors-interleukin-6 (IL-6), interleukin-16 (IL-16), tumor necrosis factor-α (TNF-α), and nuclear transcription factor-κB (NF-κB)-were measured by enzyme-linked immunosorbent assay (ELISA). (2) miRNAs capable of regulating inflammatory factors were identified through literature review and bioinformatics analysis. The regulatory relationship between miR-125b-5p and IL-6/IL-16 was verified using a dual-luciferase reporter gene assay. (3) Spinal cord neuronal cells were cultured in vitro and identified by immunofluorescence. The effects of exosomal miR-125b-5p on the expression of IL-6 and IL-16 in both SCI rat neuronal cells and spinal cord tissues were comparatively analyzed.
RESULTS: Compared with normal control group[(29.81±7.29), (35.88±5.73), (71.67±8.75), and (1 285.01±86.40) pg·mL-1, respectively], the levels of IL-6, IL-16, TNF-α and NF-κB in spinal cord tissue of model group(38.36±5.73), (46.82±7.20), (198.33±20.32), (1 907.47±120.95) pg·mL-1, respectively] increased significantly (all P<0.05). Bioinformatics search results showed that miR-125b-5p could regulate IL-6 and IL-16 simultaneously, and the results of dual luciferase reporter gene experiments confirmed this regulatory relationship directly. Cell and animal experiments have confirmed that exosomal miR-125b-5p could down regulate expression of IL-6 and IL-16.
CONCLUSION: The regulatory pathway regulating IL-6/IL-16 in spinal cord tissue through exosomal miR-125b-5p was identified in SCI rats.