Department of Anesthesiology, The General Hospital of Western Theater Command PLA, Chengdu, Sichuan 610083, P.R. China.
Department of Anesthesiology, The General Hospital of Western Theater Command PLA, Chengdu, Sichuan 610083, P.R. China.
Mol Med Rep. 2024 May;29(5). doi: 10.3892/mmr.2024.13204. Epub 2024 Mar 22.
Remifentanil‑induced hyperalgesia (RIH) is characterized by the emergence of stimulation‑induced pain, including phenomena such as allodynia and thermal hyperalgesia following remifentanil infusion. As a sequence‑specific DNA binding transcription factor, PAX6 positively and negatively regulates transcription and is expressed in multiple cell types in the developing and adult central nervous system. It was hypothesized that puerarin could relieve RIH via targeting PAX6 to regulate transcription of transient receptor potential cation channel subfamily V Member 1 (TRPV1). A total of 32 rats were randomly divided into five groups, namely control group, RI group, RI + 10 mg/kg puerarin group (RI + puerarin10), RI + 20 mg/kg puerarin group (RI + puerarin20), and RI + 40 mg/kg puerarin group (RI + puerarin40). Mechanical and thermal hyperalgesia were tested at ‑24, 2, 6, 24 and 48 h after remifentanil infusion. Following the sacrifice of rats after the last behavioral test, western blot was used to detect the expression levels of TRPV1 in the tissues; Immunofluorescence staining and western blotting were used to detect the expression of PAX6 in the spinal cord. PharmMapper and JASPAR were used to predict the binding sites of puerarin/PAX6/TRPV1. Chromatin immunoprecipitation‑PCR and dual luciferase reporter assay were used to verify the targeting relationship between PAX6 and TRPV1. Immunofluorescence was used to detect the expression levels of TRPV1 and p‑NR2B. The results revealed that puerarin (10, 20, 40 mg/kg) dose‑dependently reduced thermal and mechanical hyperalgesia from 2 to 48 h after remifentanil infusion. Remifentanil infusion remarkably stimulated the expression of phosphorylated (p‑)NR2B. Nevertheless, the increased amount of p‑NR2B by RIH was dose‑dependently suppressed by puerarin in rats. In conclusion, puerarin was revealed to attenuate postoperative RIH via targeting PAX6 to regulate the transcription of TRPV1.
瑞芬太尼诱导的痛觉过敏（RIH）的特征是出现刺激诱导的疼痛，包括瑞芬太尼输注后出现的痛觉过敏和热痛觉过敏等现象。PAX6 作为一种序列特异性 DNA 结合转录因子，可正向和负向调节转录，并在发育中和成年中枢神经系统的多种细胞类型中表达。研究假设葛根素可通过靶向 PAX6 调节瞬时受体电位阳离子通道亚家族 V 成员 1（TRPV1）的转录来缓解 RIH。将 32 只大鼠随机分为 5 组，即对照组、RI 组、RI+10mg/kg 葛根素组（RI+葛根素 10）、RI+20mg/kg 葛根素组（RI+葛根素 20）和 RI+40mg/kg 葛根素组（RI+葛根素 40）。在瑞芬太尼输注后-24、2、6、24 和 48h 时测试机械和热痛觉过敏。在最后一次行为测试后处死大鼠，采用 Western blot 检测组织中 TRPV1 的表达水平；免疫荧光染色和 Western blot 检测脊髓中 PAX6 的表达。使用 PharmMapper 和 JASPAR 预测葛根素/PAX6/TRPV1 的结合位点。使用染色质免疫沉淀-PCR 和双荧光素酶报告基因检测验证 PAX6 与 TRPV1 的靶向关系。免疫荧光检测 TRPV1 和 p-NR2B 的表达水平。结果表明，葛根素（10、20、40mg/kg）剂量依赖性地降低了瑞芬太尼输注后 2 至 48h 的热和机械痛觉过敏。瑞芬太尼输注显著刺激磷酸化（p）NR2B 的表达。然而，在大鼠中，RIH 引起的 p-NR2B 增加量被葛根素剂量依赖性地抑制。总之，葛根素通过靶向 PAX6 调节 TRPV1 的转录来减轻术后 RIH。
