miR-155通过Nrf2通路调控CCI大鼠坐骨神经病理性疼痛和炎症反应  

miR-155 regulates sciatic nerve pathological pain and inflammation in CCI rats through Nrf2 pathway

作　　者：赵飞 张弥 姚忠军[1] 朱必涛 江良波 ZHAO Fei;ZHANG Mi;YAO Zongjun;ZHU Bitao;JIANG Liangbo(Department of Orthopedics,Taihe Hospital Affiliated to Hubei University of Medicine,Shiyan 442000,China;Department of Dermatology,People's Hospital Affiliated to Hubei University of Medicine,Shiyan 442000,China;Department of Orthopedics and Traumatology,People's Hospital Affiliated to Hubei University of Medicine,Shiyan 442000,China)

机构地区：[1]湖北医药学院附属太和医院骨2科,十堰442000 [2]湖北医药学院附属人民医院皮肤科,十堰442000 [3]湖北医药学院附属人民医院创伤骨科,十堰442000

出　　处：《免疫学杂志》2021年第10期845-853,共9页Immunological Journal

摘　　要：目的明确miR-155对CCI(chronic constriction injury)介导的坐骨神经炎症反应的调控作用与机制。方法以大鼠为研究对象,建立坐骨神经CCI模型,注射miR-155 inhibitor处理。通过机械缩足阈值(paw withdrawal threshold,PWT)和热缩足潜伏期(paw withdrawal latency,PWL)评估神经疼痛行为学。qPCR检测L4~L6大鼠背侧脊髓miR-155、Nrf2、IL-6、IL-1β和TNF-α的表达。ELISA检测IL-6、IL-1β和TNF-α在近端背根神经节(L4~L6)中的水平。双荧光素酶报告基因试验验证miR-155与Nrf2的相互作用关系。Western blot检测Nrf2的蛋白表达水平。结果PWT和PWL在CCI大鼠术后1~3周显著降低,而miR-155的表达显著升高(P<0.05)。miR-155 inhibitor处理后,miR-155表达显著下调,且抑制miR-155能够上调PWT和PWL(P<0.05)。CCI处理显著升高IL-6、IL-1β和TNF-α水平,而抑制miR-155能明显逆转这一变化(P<0.05)。此外,Nrf2是miR-155的下游靶标,且降低miR-155的表达可明显提高CCI下调的Nrf2的水平(P<0.05)。沉默Nrf2能够部分逆转miR-155抑制对CCI引起的坐骨神经病理性疼痛和炎症反应的影响。结论miR-155抑制可能通过激活Nrf2缓解CCI大鼠坐骨神经炎症反应及神经疼痛,为坐骨神经慢性卡压损伤的治疗策略研发提供了新的思路。This study was performed to clarify the regulatory effect of miR-155 on chronic constriction injury(CCI)mediated sciatic nerve inflammation and its mechanism.Sciatic nerve CCI model was established in rats,and treated with miR-155 inhibitor.The ethology of neuropathic pain was evaluated by paw withdrawal threshold(PWT)and paw withdrawal latency(PWL).qPCR assay was used to detect the mRNA levels of miR-155,Nrf2,IL-6,IL-1βand TNF-αin the L4-L6 dorsal spinal cord of rats;ELISA assay was allowed to detect the levels of IL-6,IL-1βand TNF-αin the proximal dorsal root ganglion(L4-L6);the interaction of miR-155 and Nrf2 was conformed by dual luciferase reporter assay;Western blot was used to measure the protein level of Nrf2.Data showed that PWT and PWL were significantly decreased,while miR-155 expression was significantly increased in CCI rats in 1-3 weeks after operation(P<0.05),while miR-155 inhibitor could reveres theses changes(P<0.05).Furthermore,CCI remarkably increased the levels of IL-6,IL-1βand TNF-α,while inhibition of miR-155 evidently reversed these changes(P<0.05).Besides,reducing the expression of miR-155 significantly promoted CCI-induced down-regulation of Nrf2(a downstream target gene of miR-155)(P<0.05).Nrf2 silencing could partially revise effects of miR-155 inhibitor on CCI induced sciatic nerve pathological pain and inflammation(P<0.05).In conclusion,miR-155 inhibition may alleviate sciatic nerve neuropathic pain and inflammation in CCI rats by activating Nrf2,providing a new idea for the development of sciatic nerve CCI treatment strategy.

关 键 词：坐骨神经慢性卡压损伤 MIR-155 Nrf2 炎症反应 神经病理性疼痛 

分 类 号：R741.02[医药卫生—神经病学与精神病学]