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Updated: Jan 19, 2026
Chronic Constriction Injury of the Rat's Infraorbital Nerve IoN-CCI to Study Trigeminal Neuropathic Pain
Published on: September 21, 2015
miR-101 down-regulates mTOR expression and attenuates neuropathic pain in chronic constriction injury rat models
Tian Xie1, Jianrong Zhang2, Zhenming Kang3
1Department of Pain, the First Affiliated Hospital of Fujian Medical University, No. 20 Chazhong Road, Taijiang District, Fuzhou, 350005, Fujian, China.
Abstract:
We aimed to demonstrate the effects of microRNA (miR)-101 on neuropathic pain and explore the underlying mechanisms. Rat spinal microglia cells were isolated and inflammatory condition was stimulated by 24-h incubation with lipopolysaccharide (LPS). Rats were divided into 4 groups: sham, chronic constriction injury (CCI), CCI + miR-negative control (miR-NC) and CCI + miR-101 mimics. Paw withdrawal threshold (PWT) and paw withdrawal latency (PWL) tests were conducted. The mRNA levels of key genes were determined by quantitative real-time polymerase chain reaction. Mammalian target of rapamycin (mTOR) protein level was detected by Western blot. Concentrations of interleukin (IL)-6, IL-1β and tumor necrosis factor (TNF)-α were examined by ELISA. MiR-101 was downregulated and mTOR was upregulated in lumbar spinal dorsal horns from CCI rats. Targetscan and luciferase reporter assay confirmed that mTOR was direct target of miR101. MiR-101 mimics inhibited LPS-stimulated increase in the levels of IL-6, IL-1β and TNF-α in primary microglial cells in vitro. In the rat CCI model, miR-101 mimics also suppressed CCI-induced decrease in PWT and PWL and inhibited CCI-induced increase in mRNA and protein levels of IL-6, IL-1β and TNF-α. In addition, miR-101 downregulated mTOR mRNA and protein expressions in CCI rats. Besides, CCI operation resulted in miR-101 downregulation and mTOR mRNA upregulation in spinal microglia cells in a time-dependent manner. In conclusion, miR-101 had neuropathic pain-attenuating activity through targeting mTOR.
Insights
MicroRNA-101 (miR-101) alleviates neuropathic pain by targeting the mammalian target of rapamycin (mTOR) pathway. Upregulating miR-101 reduces pain behaviors and neuroinflammation, offering a potential therapeutic strategy.
Area of Science:
- Neuroscience
- Molecular Biology
- Pharmacology
Background:
- Neuropathic pain is a debilitating condition with limited treatment options.
- MicroRNAs (miRNAs) play crucial roles in regulating cellular processes, including pain signaling.
- Dysregulation of miR-101 and its target, mammalian target of rapamycin (mTOR), has been implicated in pain.
Purpose of the Study:
- To investigate the role of miR-101 in neuropathic pain.
- To explore the underlying molecular mechanisms involving mTOR.
- To assess the therapeutic potential of miR-101 mimics in a rat model of neuropathic pain.
Main Methods:
- Establishment of a chronic constriction injury (CCI) rat model.
- In vitro studies using lipopolysaccharide (LPS)-stimulated rat spinal microglia.
- Assessment of pain behaviors (paw withdrawal threshold and latency).
- Quantitative real-time polymerase chain reaction (qRT-PCR) for gene expression.
- Western blot for protein analysis.
- Enzyme-linked immunosorbent assay (ELISA) for cytokine levels.
Main Results:
- miR-101 was downregulated, while mTOR was upregulated in the spinal cords of CCI rats.
- miR-101 directly targets and downregulates mTOR.
- miR-101 mimics reduced pro-inflammatory cytokine levels (IL-6, IL-1β, TNF-α) in vitro and in vivo.
- miR-101 mimics attenuated CCI-induced pain behaviors and neuroinflammation.
- miR-101 downregulation and mTOR upregulation in spinal microglia were time-dependent following CCI.
Conclusions:
- miR-101 exhibits neuropathic pain-attenuating effects.
- The mechanism involves the direct targeting of mTOR by miR-101.
- miR-101 represents a potential therapeutic target for managing neuropathic pain.
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