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Small RNA sequencing of whole blood from a rat model of chronic primary pain with adrenergic receptor blockade

ABSTRACT: The sympathetic nervous system has been implicated in the development of chronic primary pain conditions (CPPCs), yet the downstream molecular mechanisms linking sustained catecholamine signaling to chronic pain remain poorly understood. Here, we profiled circulating small RNAs in a rat model of CPPC induced by sustained catecholamine elevation through catechol-O-methyltransferase (COMT) inhibition. Female Sprague-Dawley rats (N = 5/group) were assigned to four treatment groups: Vehicle + Vehicle (pain-free control), OR486 + Vehicle (primary pain condition), Vehicle + ICI-118,551/SR59230A, or OR486 + ICI-118,551/SR59230A. OR486 and vehicle were delivered continuously for 14 days by subcutaneous osmotic minipump. Separate minipumps delivered the ADRB2-selective antagonist ICI-118,551 and the ADRB3-selective antagonist SR59230A or vehicle. Whole blood was collected on Day 14 for small RNA isolation and Illumina small RNA sequencing. Differential expression analysis identified a discrete set of circulating miRNAs altered during primary pain, including significant downregulation of miR-133a-3p and upregulation of multiple inflammation-associated miRNAs. Pharmacologic blockade of ADRB2/3 prevented miR-133a-3p downregulation, identifying miR-133a-3p as an adrenergic receptor-dependent regulator associated with chronic primary pain.

ORGANISM(S): Rattus norvegicus

PROVIDER: GSE338452 | GEO | 2026/07/13

REPOSITORIES: GEO

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Project description:We profiled the expression of circulating microRNAs (miRNAs) in mice exposed to gram-positive and gram-negative bacteria using Illumina small RNA deep sequencing. Recombinant-specific gram-negative pathogen Escherichia coli (Xen14) and gram-positive pathogen Staphylococcus aureus (Xen29) were used to induce bacterial infection in mice at a concentration of 1 × 108 bacteria/100 μL of phosphate buffered saline (PBS). Small RNA libraries generated from the serum of mice after exposure to PBS, Xen14, Xen29, and Xen14+Xen29 via the routes of subcutaneous injection (I), cut wound (C), or under grafted skin (S) were analyzed using an Illumina HiSeq2000 Sequencer. Following exposure to gram-negative bacteria alone, no differentially expressed miRNA was found in the injection, cut, or skin graft models. Exposure to mixed bacteria induced a similar expression pattern of the circulating miRNAs to that induced by gram-positive bacterial infection. Upon gram-positive bacterial infection, 9 miRNAs (mir-193b-3p, mir-133a-1-3p, mir-133a-2-3p, mir-133a-1-5p, mir-133b-3p, mir-434-3p, mir-127-3p, mir-676-3p, mir-215-5p) showed upregulation greater than 4-fold with a p-value < 0.01. Among them, mir-193b-3p, mir-133a-1-3p, and mir-133a-2-3p presented the most common miRNA targets expressed in the mice exposed to gram-positive bacterial infection. Male C57BL/6 mice (age, 10–12 weeks; weight, 30–35 g) were purchased from BioLasco (Yi-Lan, Taiwan). The mice were anesthetized by intraperitoneal injection of an anesthetic cocktail consisting of 0.1 mg/g ketamine and 0.01 mg/g xylazine. The anesthetized mice were restrained in a supine position on a heated pad to maintain body temperature at 37°C. Recombinant-specific gram-negative pathogen Escherichia coli (Xen14) and gram-positive pathogen Staphylococcus aureus (Xen29) purchased from Caliper (Caliper, USA) were used to induce bacterial infection in the mice at a concentration of 1 × 108 bacteria/100 μL of phosphate buffered saline (PBS). To create mixed gram-negative and gram-positive bacterial infection, 1 × 108 Xen14 bacteria and 1 × 108 Xen29 bacteria/100 μL of PBS were used for wound contamination. Three animal models were used to create bacterial infection routes: subcutaneous injection (hereafter referred to as (I)), cut wound (hereafter referred to as (C)), and skin grafting (hereafter referred to as (S)). In the (I) model, E. coli and/or S. aureus suspensions were injected subcutaneously into the backs of the mice using an Fr. 25 needle. In the (C) model, a 1 cm incision wound was created in the midline of the back, smeared with E. coli and/or S. aureus suspension, and the wound was closed directly with a 4-0 nylon suture. In the (S) model, a 1×1 cm rectangular full- thickness skin graft was lifted from the backs of the mice, E. coli and/or S. aureus suspensions were spread over the wound bed, and the skin graft was reattached and closed with a 4-0 nylon suture. An additional group of animals in each of these three models was inoculated with PBS to serve as a negative control. Small RNA libraries generated from the serum of mice after exposure to PBS, Xen14, Xen29, and Xen14+Xen29 via the routes of subcutaneous injection (I), cut wound (C), or under grafted skin (S) were analyzed using an Illumina HiSeq2000 Sequencer.

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Small RNA sequencing of whole blood from a rat model of chronic primary pain with adrenergic receptor blockade

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