A growing body of studies have indicated that bone marrow mesenchymal stem cells (BMSCs) have powerful analgesic effects in animal models of bone cancer pain. Here, we explored the molecular mechanisms underlying how BMSCs alleviate pain sensation in a mouse model of bone cancer pain. C3H/HeN adult male mice were used to generate a bone cancer pain model. BMSCs were isolated from mouse bone marrow, modified by transfection with microRNA-9-5p (miR-9-5p), and infused into the spinal cord. Spontaneous flinches, paw withdrawal latency, limb-use score, and weight-bearing score were used to assess pain-related behaviors. ELISA, RT-PCR, western blot, and luciferase assay were used to assess gene expressions. Our results show that miR-9-5p regulated the expression of both repressor element silencing transcription factor (REST) and μ-opioid receptors (MOR) by targeting REST in primary mouse BMSCs. Overexpression of miR-9-5p reversed the activation of inflammatory pathway in TNF-α- and IL-6-treated BMSCs. In addition, miR-9-5p modified BMSCs alleviated cancer pain in the sarcoma-inoculated mouse model. MiR-9-5p modified BMSCs suppressed cytokine expression in the spinal cord of sarcoma-inoculated mice by suppressing REST gene expression. Our results indicate that miR-9-5p modified BMSCs can relieve bone cancer pain via modulating neuroinflammation in the central nervous system, suggesting genetically modified BMSCs could be a promising cell therapy in pain management.

中文翻译：

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鞘内注射miR-9-5p修饰的小鼠骨髓间充质干细胞对骨癌痛小鼠模型的镇痛作用

越来越多的研究表明，骨髓间充质干细胞（BMSC）在骨癌疼痛的动物模型中具有强大的镇痛作用。在这里，我们探讨了骨髓间充质干细胞如何缓解骨癌疼痛小鼠模型中的疼痛感的分子机制。C3H / HeN成年雄性小鼠用于生成骨癌疼痛模型。从小鼠骨髓中分离骨髓间充质干细胞，用microRNA-9-5p（miR-9-5p）转染修饰，然后注入脊髓。使用自发性退缩，爪退缩潜伏期，肢体使用评分和负重评分来评估疼痛相关行为。ELISA，RT-PCR，蛋白质印迹和荧光素酶测定法用于评估基因表达。我们的研究结果表明，miR-9-5p通过靶向原代小鼠BMSC中的REST来调节阻遏元件沉默转录因子（REST）和μ阿片受体（MOR）的表达。miR-9-5p的过表达逆转了TNF-α和IL-6处理的BMSC中炎症途径的激活。此外，miR-9-5p修饰的BMSC可减轻肉瘤接种小鼠模型的癌痛。MiR-9-5p修饰的BMSC通过抑制REST基因表达来抑制肉瘤接种小鼠脊髓中细胞因子的表达。我们的结果表明，miR-9-5p修饰的BMSC可以通过调节中枢神经系统的神经炎症来缓解骨癌疼痛，这表明基因修饰的BMSC可能是一种有希望的疼痛治疗细胞疗法。miR-9-5p的过表达逆转了TNF-α和IL-6处理的BMSC中炎症途径的激活。此外，miR-9-5p修饰的BMSC可减轻肉瘤接种小鼠模型的癌痛。MiR-9-5p修饰的BMSC通过抑制REST基因表达来抑制肉瘤接种小鼠脊髓中细胞因子的表达。我们的结果表明，miR-9-5p修饰的BMSC可以通过调节中枢神经系统的神经炎症来缓解骨癌疼痛，这表明基因修饰的BMSC可能是一种有希望的疼痛治疗细胞疗法。miR-9-5p的过表达逆转了TNF-α和IL-6处理的BMSC中炎症途径的激活。此外，miR-9-5p修饰的BMSC可减轻肉瘤接种小鼠模型的癌痛。MiR-9-5p修饰的BMSC通过抑制REST基因表达来抑制肉瘤接种小鼠脊髓中细胞因子的表达。我们的结果表明，miR-9-5p修饰的BMSC可以通过调节中枢神经系统的神经炎症来缓解骨癌疼痛，这表明基因修饰的BMSC可能是一种有希望的疼痛治疗细胞疗法。MiR-9-5p修饰的BMSC通过抑制REST基因表达来抑制肉瘤接种小鼠脊髓中细胞因子的表达。我们的结果表明，miR-9-5p修饰的BMSC可以通过调节中枢神经系统的神经炎症来缓解骨癌疼痛，这表明基因修饰的BMSC可能是一种有希望的疼痛治疗细胞疗法。MiR-9-5p修饰的BMSC通过抑制REST基因表达来抑制肉瘤接种小鼠脊髓中细胞因子的表达。我们的结果表明，miR-9-5p修饰的BMSC可以通过调节中枢神经系统的神经炎症来缓解骨癌疼痛，这表明基因修饰的BMSC可能是一种有希望的疼痛治疗细胞疗法。