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脊髓lncRNA_RT1-CE10通过抑制小胶质细胞的活化缓解肠易激综合征大鼠内脏痛觉敏化

刘姿含^1,2, 刘艺倩², 张倩丽², 林春², 黄扬^2,*

¹平顶山学院医学院，平顶山 467000；²福建医科大学基础医学院疼痛研究所，福州 350122

摘要

This study aimed to investigate the role of 5-hydroxytryptamine 1A (5-HT1A) receptors in the anterior cingulate cortex (ACC) in regulating the affective pain in rats. In the ACC, normal saline (NS), the 5-HT1A receptor agonist 8-OH-DPAT or the 5-HT1A receptor antagonist WAY-100635 was pre-injected respectively. A persistent inflammatory pain model was established by subcutaneously injecting 0.08 mL of complete Freund's adjuvant (CFA) into the left hind paw of rats, which was then paired with a specific environment to induce a conditioned place avoidance (CPA) response. Then the electroacupuncture (EA, 10 Hz, 3 mA) was applied to stimulate the Huantiao acupoint (GB30). The CPA responses of rats in the pain-paired environment and the firing activities of ACC neurons were simultaneously observed. Subsequently, open-field behavior, paw withdrawal latency (PWL), and 50% paw withdrawal threshold (PWT) tests were conducted. The results showed that: (1) Rats injected with CFA showed significant reductions in PWL and 50% PWT, and spent less time in the "pain-paired environment" and the center of the open field, compared to the control group (P < 0.05). (2) Immunofluorescence double-labeling results showed a high co-expression of Fos protein and 5-HT1A receptors in the ACC of the EA-treated normal rats. (3) Pre-treatment with the 5-HT1A receptor agonist 8-OH-DPAT (8 μg) in the ACC alleviated CFA-induced affective pain (P < 0.05) and reversed the increase in firing frequency of pyramidal neurons in the ACC induced by CFA (P < 0.05). (4) Pre-treatment with the 5-HT1A receptor antagonist WAY-100635 (8 μg) in the ACC, combined with EA stimulation, reversed the CPA-like behavioral responses induced by CFA in rats and increased the firing frequency of pyramidal neurons in the ACC (P < 0.05). These results suggest that the 5-HT1A receptors in ACC mediate the alleviating effect of EA on the affective pain of CFA-induced rats. 本文旨在探究脊髓lncRNA_RT1-CE10在肠易激综合征(irritable bowel syndrome, IBS)大鼠慢性功能性内脏痛中枢敏化中的作用机制。采用新生儿期结直肠扩张(colorectal distention, CRD)刺激的方法建立IBS 大鼠模型，通过记录6 周龄时腹外斜肌在CRD (40/60 mmHg) 下的肌电活动评估大鼠内脏痛的痛觉敏化。采用RT-qPCR 检测大鼠胸腰段脊髓背角lncRNA_RT1-CE10 的表达水平。通过Western blot 检测大鼠脊髓背角Iba-1 和MHCII的表达以评估小胶质细胞活化状态，比较鞘内注射米诺环素(小胶质细胞特异性抑制剂)前后IBS 大鼠内脏痛觉敏化程度及小胶质细胞活化水平。此外，通过鞘内注射AAV-lncRT1-CE10 或AAV-shlncRT1-CE10 分别实现lncRNA_RT1-CE10 的过表达与敲低，并观察其对小胶质细胞活化的影响。结果显示：IBS大鼠脊髓背角lncRNA_RT1-CE10 表达显著降低。同时，IBS大鼠脊髓背角小胶质细胞被激活，抑制小胶质细胞的活化可缓解IBS 大鼠的内脏痛觉敏化。过表达lncRNA_RT1-CE10 可抑制IBS 大鼠脊髓背角小胶质细胞的活化；相反，敲低lncRNA_RT1-CE10 的表达则可激活正常大鼠脊髓背角小胶质细胞。上述结果提示，脊髓lncRNA_RT1-CE10 可通过抑制小胶质细胞的活化缓解IBS大鼠慢性功能性内脏痛的痛觉敏化。

关键词： 肠易激综合征; 长链非编码RNA; 小胶质细胞; 痛觉敏化; 脊髓

Spinal lncRNA_RT1-CE10 attenuates chronic visceral hypersensitivity in rats with irritable bowel syndrome via inhibiting microglial activation

LIU Zi-Han^1,2, LIU Yi-Qian², ZHANG Qian-Li², LIN Chun², HUANG Yang^2,*

¹College of Medicine, Pingdingshan University, Pingdingshan 467000, China；²Institute of Pain Research, School of Basic Medical Sciences, Fujian Medical University, Fuzhou 350122, China

Abstract

The purpose of the study was to investigate the role of the long non-coding RNA (lncRNA)_RT1-CE10 and its molecular mechanisms in chronic visceral pain of rats with irritable bowel syndrome (IBS). Sprague-Dawley (SD) rats were exposed to colorectal distention (CRD) stimulation at a pressure of 60 mmHg for 1 min from 8 to 14 days after birth. The visceral hypersensitivity was assessed by measuring electromyographic (EMG) responses of external oblique muscle to CRD at 40 mmHg and 60 mmHg when the rats were 6 weeks old. The expression of lncRNA_RT1-CE10, Iba-1 and MHCII in the spinal cord was detected by RT-qPCR or Western blot. After intrathecal injection of minocycline (an inhibitor of microglia), the expression of Iba-1, MHCII and the visceral hypersensitivity were determined by Western blot and EMG, respectively. After intrathecal injection of AAV-lncRT1-CE10 and AAVshlncRT1- CE10, the expression of Iba-1 and MHCII was examined by Western blot, respectively. The results showed that the expression of lncRNA_RT1-CE10 was decreased in the spinal cord of IBS rats, while the expression of Iba-1 and MHCII was increased. Inhibition of microglial activation by minocycline attenuated visceral hypersensitivity in IBS rats. Over-expression of lncRNA_RT1- CE10 decreased the Iba-1 and MHCⅡ levels in IBS rats, while knockdown of lncRNA_RT1-CE10 increased the Iba-1 and MHCⅡ levels in control rats. Collectively, these results demonstrate that lncRNA_RT1-CE10 attenuates visceral hypersensitivity by inhibiting microglial activation in IBS rats.

Key words: irritable bowel syndrome; lncRNA; microglia; hyperalgesia; spinal cord

收稿日期：　　录用日期：

通讯作者：黄扬　　E-mail:

DOI: 10.13294/j.aps.2026.0002

引用本文：

刘姿含, 刘艺倩, 张倩丽, 林春, 黄扬. 脊髓lncRNA_RT1-CE10通过抑制小胶质细胞的活化缓解肠易激综合征大鼠内脏痛觉敏化[J]. 生理学报 2026; 78 (1): 246-252.

LIU Zi-Han, LIU Yi-Qian, ZHANG Qian-Li, LIN Chun, HUANG Yang. Spinal lncRNA_RT1-CE10 attenuates chronic visceral hypersensitivity in rats with irritable bowel syndrome via inhibiting microglial activation. Acta Physiol Sin 2026; 78 (1): 246-252 (in Chinese with English abstract).