Department of Anesthesiology, State Key Laboratory of Oncology in South China, Guangdong Provincial Clinical Research Center for Cancer, Sun Yat-Sen University Cancer Center, Guangzhou, 510060, P. R. China.
Guangdong Province Key Laboratory of Brain Function and Disease, Department of Physiology, Zhongshan School of Medicine, Sun Yat-Sen University, Guangzhou, China.
BMC Med. 2024 Sep 20;22(1):406. doi: 10.1186/s12916-024-03634-2.
Physical symptoms and aversion induced by opioid withdrawal strongly affect the management of opioid addiction. YTH N6-methyladenosine (mA) RNA binding protein 1 (YTHDF1), an mA-binding protein, from the periaqueductal gray (PAG) reportedly contributes to morphine tolerance and hyperalgesia. However, the role of YTHDF1 in morphine withdrawal remains unclear.
A naloxone-precipitated morphine withdrawal model was established in C57/BL6 mice or transgenic mice. YTHDF1 was knocked down via adeno-associated virus transfection. Combined with the results of the single-cell RNA sequencing analysis, the changes in morphine withdrawal somatic signs and conditioned place aversion (CPA) scores were compared when YTHDF1 originating from different neurons in the ventrolateral periaqueductal gray (vlPAG) was knocked down. We further explored the role of inflammatory factors and transcription factors related to inflammatory response in morphine withdrawal.
Our results revealed that YTHDF1 expression was upregulated in the vlPAG of mice with morphine withdrawal and that the knockdown of vlPAG YTHDF1 attenuated morphine withdrawal-related somatic signs and aversion. The levels of NF-κB and p-NF-κB were reduced after the inhibition of YTHDF1 in the vlPAG. YTHDF1 from vlPAG inhibitory neurons, rather than excitatory neurons, facilitated morphine withdrawal responses. The inhibition of YTHDF1 in vlPAG somatostatin (Sst)-expressing neurons relieved somatic signs of morphine withdrawal and aversion, whereas the knockdown of YTHDF1 in cholecystokinin (Cck)-expressing or parvalbumin (PV)-expressing neurons did not change morphine withdrawal-induced responses. The activity of c-fos + neurons, the intensity of the calcium signal, the density of dendritic spines, and the frequency of mIPSCs in the vlPAG, which were increased in mice with morphine withdrawal, were decreased with the inhibition of YTHDF1 from vlPAG inhibitory neurons or Sst-expressing neurons. Knockdown of NF-κB in Sst-expressing neurons also alleviated morphine withdrawal-induced responses.
YTHDF1 originating from Sst-expressing neurons in the vlPAG is crucial for the modulation of morphine withdrawal responses, and the underlying mechanism might be related to the regulation of the expression and phosphorylation of NF-κB.
阿片类药物戒断引起的躯体症状和厌恶感强烈影响阿片类药物成瘾的管理。来自导水管周围灰质（PAG）的 YTH N6-甲基腺苷（mA）RNA 结合蛋白 1（YTHDF1）是一种 mA 结合蛋白，据报道，它有助于吗啡耐受和痛觉过敏。然而，YTHDF1 在吗啡戒断中的作用尚不清楚。
在 C57/BL6 小鼠或转基因小鼠中建立纳洛酮诱发的吗啡戒断模型。通过腺相关病毒转染敲低 YTHDF1。结合单细胞 RNA 测序分析的结果，比较了当敲低腹外侧导水管周围灰质（vlPAG）中不同神经元来源的 YTHDF1 时，吗啡戒断躯体症状和条件位置厌恶（CPA）评分的变化。我们进一步探讨了与炎症反应相关的炎症因子和转录因子在吗啡戒断中的作用。
我们的结果显示，吗啡戒断小鼠 vlPAG 中 YTHDF1 表达上调，vlPAG YTHDF1 敲低可减轻吗啡戒断相关的躯体症状和厌恶。vlPAG 中 YTHDF1 抑制后 NF-κB 和 p-NF-κB 水平降低。vlPAG 抑制性神经元中的 YTHDF1 而不是兴奋性神经元促进吗啡戒断反应。vlPAG 中生长抑素（Sst）表达神经元中 YTHDF1 的抑制减轻了吗啡戒断引起的躯体症状和厌恶，而 Cck 表达或钙调蛋白（PV）表达神经元中 YTHDF1 的敲低并未改变吗啡戒断引起的反应。吗啡戒断小鼠 vlPAG 中 c-fos+神经元的活性、钙信号强度、树突棘密度和 mIPSCs 的频率增加，vlPAG 抑制性神经元或 Sst 表达神经元中 YTHDF1 的抑制降低了这些变化。Sst 表达神经元中 NF-κB 的敲低也减轻了吗啡戒断引起的反应。
vlPAG 中 Sst 表达神经元来源的 YTHDF1 对于调节吗啡戒断反应至关重要，其潜在机制可能与 NF-κB 的表达和磷酸化调节有关。
