Spinal Neuronal GRK2 Contributes to Preventive Effect by Electroacupuncture on Cisplatin-Induced Peripheral Neuropathy in Mice.
Spinal Neuronal GRK2 Contributes to Preventive Effect by Electroacupuncture on Cisplatin-Induced Peripheral Neuropathy in Mice.
复制标题
脊髓神经元 GRK2 有助于电针预防顺铂引起的小鼠周围神经病变
DOI：
10.1213/ane.0000000000005768
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发表时间：
2022-01-01
影响因子：
5.7
通讯作者：
Mao-Ying QL
中科院分区：
文献类型：
--
作者：
Ma X;Chen Y;Li XC;Mi WL;Chu YX;Wang YQ;Mao-Ying QL
化疗引起的周围神经病变（CIPN）的主要症状包括疼痛和麻木。神经元G蛋白偶联受体激酶2（GRK 2）在多种疼痛模型中起着重要作用。顺铂治疗可诱导脊髓中促炎性小胶质细胞活化。本研究的目的是探讨脊髓神经元GRK 2在顺铂诱导的CIPN中的作用以及在电针预防CIPN中的作用。采用vonFrey实验和粘连清除实验检测小鼠的疼痛和感觉缺失行为。脊髓中神经元GRK 2的表达通过脊髓内注射含有神经元特异性启动子的腺相关病毒（AAV）来调节。Western blot检测脊髓背角GRK 2、髓样细胞触发受体2（TREM 2）和12 kDa DNA X激活蛋白（DAP 12）的蛋白水平，免疫荧光法检测表皮内神经纤维（IENF）密度，实时荧光PCR法检测小胶质细胞活化情况。在本研究中，顺铂治疗导致脊髓背角GRK 2表达减少。脊髓内注射带有人突触蛋白（hSyn）启动子的GRK 2腺相关病毒载体，使GRK 2在脊髓内过表达，可显著抑制内神经纤维的丢失，减轻顺铂引起的机械性疼痛和感觉障碍。Real-time PCR分析显示，神经元GRK 2的过表达显著抑制顺铂诱导的促炎细胞因子白细胞介素（IL）-1β、IL-6、诱导型一氧化氮合酶（iNOS）和M1小胶质细胞标志物分化簇（CD）16的信使RNA（mRNA）上调。此外，在本研究中，已被证明在小胶质细胞激活和CIPN发育中发挥作用的TREM 2和DAP 12也因神经元GRK 2的过表达而下调。有趣的是，在顺铂诱导的CIPN小鼠模型中，EA的预防性治疗完全模拟了脊髓中神经元GRK 2过表达的效果。电针增加顺铂治疗后脊髓背角GRK 2的表达。脊髓内注射AAV载体特异性下调神经元GRK 2，完全逆转EA对CIPN和小胶质细胞活化的调节作用。提示神经元GRK 2介导的小胶质细胞活化参与了CIPN的形成过程。脊髓神经元GRK 2参与了电针对CIPN的预防作用。神经元GRK 2可能是CIPN干预的潜在靶点。
The main symptoms of chemotherapy-induced peripheral neuropathy (CIPN) include pain and numbness. Neuronal G protein–coupled receptor kinase 2 (GRK2) plays an important role in various pain models. Cisplatin treatment can induce the activation of proinflammatory microglia in spinal cord. The purpose of this study was to investigate the role of spinal neuronal GRK2 in cisplatin-induced CIPN and in the prevention of CIPN by electroacupuncture (EA). The pain and sensory deficit behaviors of mice were examined by von Frey test and adhesive removal test. The expression of neuronal GRK2 in the spinal cord is regulated by intraspinal injection of adeno-associated virus (AAV) containing neuron-specific promoters. The protein levels of GRK2, triggering receptor expressed on myeloid cells 2 (TREM2), and DNAX-activating protein of 12 kDa (DAP12) in spinal dorsal horn were detected by Western blot, the density of intraepidermal nerve fibers (IENFs) was detected by immunofluorescence, and microglia activation were evaluated by real-time polymerase chain reaction (PCR). In this study, cisplatin treatment led to the decrease of GRK2 expression in the dorsal horn of spinal cord. Overexpression of neuronal GRK2 in spinal cord by intraspinal injection of an AAV vector expressing GRK2 with human synapsin (hSyn) promotor significantly inhibited the loss of IENFs and alleviated the mechanical pain and sensory deficits induced by cisplatin. Real-time PCR analysis showed that the overexpression of neuronal GRK2 significantly inhibited the messenger RNA (mRNA) upregulation of proinflammatory cytokine interleukin (IL)-1β, IL-6, inducible nitric oxide synthase (iNOS), and M1 microglia marker cluster of differentiation (CD)16 induced by cisplatin. Furthermore, the TREM2 and DAP12, which has been demonstrated to play a role in microglia activation and in the development of CIPN, were also downregulated by overexpression of neuronal GRK2 in this study. Interestingly, preventive treatment with EA completely mimics the effect of overexpression of neuronal GRK2 in the spinal cord in this mouse model of cisplatin-induced CIPN. EA increased GRK2 level in spinal dorsal horn after cisplatin treatment. Intraspinal injection of AAV vector specifically downregulated neuronal GRK2, completely reversed the regulatory effect of EA on CIPN and microglia activation. All these indicated that the neuronal GRK2 mediated microglial activation contributed to the process of CIPN. Neuronal GRK2 in the spinal cord contributed to the preventive effect of EA on CIPN. The neuronal GRK2 may be a potential target for CIPN intervention.