PIRB在糖尿病周围神经病理性疼痛中的作用及其调控机制

The Role and Regulatory Mechanisms of PIRB in Diabetic Peripheral Neuropathic Pain

摘要:

目的:本研究旨在探讨配对免疫球蛋白受体B(PIRB)在糖尿病周围神经病理性疼痛(DPNP)中的作用及其潜在机制,特别关注其在神经炎症和炎性小体活化中的作用。方法:本实验将小鼠随机分为4组:假手术组(sham)、模型组 (DPNP)、模型+PIRB阴性对照组 (DPNP + AAV control)、模型+PIRB腺相关病毒组(DPNP + PIRB)。采用腹腔注射链脲佐菌素(stz)构建小鼠的糖尿病周围神经病理性疼痛模型,并通过血糖升高及机械性痛觉过敏确认模型的成功建立。使用免疫印迹和qRT-PCR评估DPNP小鼠脊髓组织中PIRB的表达水平。通过鞘内注射腺相关病毒(AAV)降低小鼠脊髓中PIRB的表达以进一步研究其作用。采用qRT-PCR检测神经炎症标志物(如IL-1β、IL-6、TNF-α),使用免疫印迹法检测NF-κB p65的水平,并通过免疫印迹和免疫组化观察NLRP3炎性小体成分的变化。结果:与sham组相比,DPNP组小鼠出现显著机械性痛觉过敏,并观察到PIRB的蛋白和mRNA表达水平均显著上调(p < 0.05)。通过AAV介导的PIRB敲低实验中,结果显示DPNP+PIRB组小鼠机械性痛觉过敏减轻(p < 0.05),并且神经炎症标志物(IL-1β、IL-6、TNF-α)的表达减少。此外,DPNP+PIRB组较DPNP+AAV control组的NF-κB p65和NLRP3炎性小体成分(如Caspase-1、GSDMD和IL-1β)的表达也显著降低。结论:PIRB可能通过调节神经炎症和激活NLRP3炎性小体在糖尿病周围神经病理性疼痛的发展中发挥了关键作用,抑制PIRB的表达可以减轻痛觉过敏现象并减少神经炎症。

Abstract:

Objective: This study aims to investigate the role of paired immunoglobulin-like receptor B (PIRB) in diabetic peripheral neuropathic pain (DPNP) and its potential mechanisms, with a specific focus on its involvement in neuroinflammation and inflammasome activation.Methods: Mice were randomly divided into four groups: Sham group, diabetic peripheral neuropathic pain model group (DPNP), DPNP + PIRB negative control group (DPNP + AAV control), and DPNP + PIRB adeno-associated virus group (DPNP + PIRB). A diabetic peripheral neuropathic pain (DPNP) model was induced by intraperitoneal injection of streptozotocin (STZ), and the successful establishment of the model was confirmed through elevated blood glucose levels and mechanical hyperalgesia. PIRB expression levels in the spinal cord of DPNP mice were assessed using Western blot and qRT-PCR. To further investigate the role of PIRB, its expression in the spinal cord was knocked down via intrathecal injection of adeno-associated virus (AAV). Neuroinflammatory markers, such as IL-1β, IL-6, and TNF-α, were detected using qRT-PCR, while NF-κB p65 levels were evaluated by Western blot. Changes in components of the NLRP3 inflammasome were observed through both Western blot and immunohistochemistry.Results: Compared to the Sham group, the DPNP group exhibited significant mechanical hyperalgesia, along with a marked upregulation of PIRB protein and mRNA expression (p < 0.05). In the AAV-mediated PIRB knockdown experiment, the DPNP + PIRB group showed reduced mechanical hyperalgesia (p < 0.05) and decreased expression of neuroinflammatory markers (IL-1β, IL-6, TNF-α). Furthermore, compared to the DPNP + AAV control group, the DPNP + PIRB group exhibited significantly lower levels of NF-κB p65 and NLRP3 inflammasome components, including Caspase-1, GSDMD, and IL-1β.Conclusion: PIRB may play a key role in the development of diabetic peripheral neuropathic pain by regulating neuroinflammation and activating the NLRP3 inflammasome. Inhibiting PIRB expression can alleviate hyperalgesia and reduce neuroinflammation.