推拿对坐骨神经慢性压迫损伤大鼠脊髓背角小胶质细胞P2Y12/RhoA/ROCK2通路及c-Fos蛋白表达的影响OACSTPCD
Effects of Tuina on P2Y12/RhoA/ROCK2 Pathway and c-Fos Protein Expression of Microglia in Spinal Dorsal Horn of Sciatic Nerve Chronic Compressive Injury Rats
目的 观察推拿对坐骨神经慢性压迫损伤大鼠脊髓背角小胶质细胞P2Y12/RhoA/ROCK2通路及c-Fos蛋白表达的影响,探讨推拿治疗腰椎间盘突出症的作用机制.方法 采用右侧坐骨神经慢性压迫损伤模拟腰椎间盘突出症神经性疼痛.将24只雄性SD大鼠随机分为空白组、模型组和推拿组,每组8只.造模后第4日,推拿组以按揉法干预,连续14 d.测量造模前及造模后第4、10、17 d大鼠机械缩足阈值(PWT)、热痛阈值(PWL),免疫荧光染色检测大鼠右侧脊髓背角Iba1、P2Y12蛋白表达,Western blot检测右侧脊髓背角RhoA、ROCK2蛋白表达,免疫组化检染色测右侧脊髓背角c-Fos阳性细胞数量.结果 与空白组比较,模型组大鼠造模后第4、10、17日PWT、PWL明显降低(P<0.001),右侧脊髓背角Iba1、P2Y12、RhoA、ROCK2蛋白表达明显升高(P<0.001,P<0.05),c-Fos阳性细胞数明显增加(P<0.001);与模型组比较,推拿组大鼠造模后第 10、17 日PWT、PWL明显升高(P<0.05,P<0.01,P<0.001),右侧脊髓背角Iba1、P2Y12、RhoA、ROCK2蛋白表达明显降低(P<0.001,P<0.01,P<0.05),c-Fos阳性细胞数明显减少(P<0.001).结论 推拿可能通过调控脊髓背角P2Y12/RhoA/ROCK2通路及c-Fos表达抑制小胶质细胞激活,降低神经元兴奋性,对腰椎间盘突出症发挥镇痛作用.
Objective To observe the effects of tuina on P2Y12/RhoA/ROCK2 pathway and c-Fos protein expression of microglia in spinal dorsal horn of sciatic nerve chronic compressive injury(CCI)rats;To explore the mechanism of tuina in treating lumbar disc herniation.Methods A CCI model of right sciatic nerve was used to simulate neuropathic pain in lumbar disc herniation.24 male SD rats were randomly divided into blank group,model group and tuina group,with 8 rats in each group.After 4 days of modeling,massage intervention was used to in the tuina group for 14 days.The paw withdrawal threshold(PWT)and paw withdrawal latency(PWL)of rats before and on the 4th,10th,and 17th day after modeling were observed,immunofluorescence staining was used to detect the expression of Iba1 and P2Y12 protein in right spinal dorsal horn,Western blot was used to detect the expressions of RhoA and ROCK2 protein in right spinal dorsal horn,immunohistochemistry staining was used to detect the number of c-Fos positive cells in right spinal dorsal horn.Results Compared with the blank group,the PWT and PWL of the model group were significantly reduced on the 4th,10th,and 17th day after modeling(P<0.001),while the expressions of Iba1,P2Y12,RhoA and ROCK2 protein in right spinal dorsal horn significantly increased(P<0.001,P<0.05),the number of c-Fos positive cells significantly increased(P<0.001).Compared with the model group,the PWT and PWL of the tuina group significantly increased on the 10th and 17th day after modeling(P<0.05,P<0.01,P<0.001),and the expressions of Iba1,P2Y12,RhoA and ROCK2 protein in right spinal dorsal horn significantly decreased(P<0.001,P<0.01,P<0.05),and the number of c-Fos positive cells significantly reduced(P<0.001).Conclusion Tuina may inhibit the activation of microglia by regulating P2Y12/RhoA/ROCK2 pathway and c-Fos protein expression in spinal dorsal horn,reduce neuronal excitability,and exert analgesic effects on lumbar disc herniation.
蒋晶晶;林志刚;黄红叶;张幻真;陈乐春;林惠;伍诗烨;陈水金
福建中医药大学附属康复医院,福建 福州 350003||福建省康复技术重点实验室,福建 福州 350003福建中医药大学,福建 福州 350100
中医学
腰椎间盘突出症推拿脊髓背角小胶质细胞P2Y12/RhoA/ROCK2通路c-Fos大鼠
lumbar disc herniationtuinaspinal dorsal hornmicrogliaP2Y12/RhoA/ROCK2 pathwayc-Fosrats
《中国中医药信息杂志》 2024 (007)
100-105 / 6
国家自然科学基金青年科学基金(82205303、82105039);国家自然科学基金面上项目(82174523);福建省自然科学基金面上项目(2023J01878、2020J01760);福建省卫健委医学创新课题(2020CXA052);福建省卫健委中青年骨干人才培养项目(2021GGA060)
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