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Thymosin Beta 4 Is Involved in the Development of Electroacupuncture Tolerance.

ABSTRACT: Background: Electroacupuncture (EA) tolerance, a negative therapeutic effect, is a gradual decline in antinociception because of its repeated or prolonged use. This study aims to explore the role of thymosin beta 4 (T?4), having neuro-protection properties, in EA tolerance (EAT). Methods: Rats were treated with EA once daily for eight consecutive days to establish EAT, effect of T?4 on the development of EAT was determined through microinjection of T?4 antibody and siRNA into the cerebroventricle. The mRNA and protein expression profiles of T?4, opioid peptides (enkephalin, dynorphin and endorphin), and anti-opioid peptides (cholecystokinin octapeptide, CCK-8 and orphanin FQ, OFQ), and mu opioid receptor (MOR) and CCK B receptor (CCKBR) in the brain areas (hypothalamus, thalamus, cortex, midbrain and medulla) were characterized after T?4 siRNA was administered. Results: T?4 levels were increased at day 1, 4, and 8 and negatively correlated with the changes of tail flick latency in all areas. T?4 antibody and siRNA postponed EAT. T?4 siRNA caused decreased T?4 levels in all areas, which resulted in increased enkephalin, dynorphin, endorphin and MOR levels in most measured areas during repeated EA, but unchanged OFQ, CCK-8, and CCKBR levels in most measured areas. T?4 levels were negatively correlated with enkephalin, dynorphin, endorphin, or MOR levels in all areas except medulla, while positively correlated with OFQ and CCK-8 levels in some areas. Conclusion: These results confirmed T?4 facilitates EAT probably through negatively changing endogenous opioid peptides and their receptors and positively influencing anti-opioid peptides in the central nervous system.

SUBMITTER: Wan J

PROVIDER: S-EPMC6444270 | biostudies-literature | 2019

REPOSITORIES: biostudies-literature

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Thymosin Beta 4 Is Involved in the Development of Electroacupuncture Tolerance.

Wan Juan J Ding Yi Y Nan Sha S Zhang Qiulin Q Sun Jinrui J Suo Chuanguang C Ding Mingxing M

Frontiers in cellular neuroscience 20190326

<b>Background:</b> Electroacupuncture (EA) tolerance, a negative therapeutic effect, is a gradual decline in antinociception because of its repeated or prolonged use. This study aims to explore the role of thymosin beta 4 (Tβ4), having neuro-protection properties, in EA tolerance (EAT). <b>Methods:</b> Rats were treated with EA once daily for eight consecutive days to establish EAT, effect of Tβ4 on the development of EAT was determined through microinjection of Tβ4 antibody and siRNA into the c ...[more]

PMID: 30971892

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Project description:Thymosin beta-4 (T?4) is an ubiquitous multi-functional regenerative peptide, related to many critical biological processes, with a dynamic and flexible conformation which may influence its functions and its subcellular distribution. For these reasons, the intracellular localization and trafficking of T?4 is still not completely defined and is still under investigation in in vivo as well as in vitro studies. In the current study we used HepG2 cells, a human hepatoma cell line; cells growing in normal conditions with fetal bovine serum expressed high levels of T?4, restricted to the cytoplasm until 72 h. At 84 h, a diffuse T?4 cytoplasmic immunostaining shifted to a focal perinuclear and nuclear reactivity. In the absence of serum, nuclear reactivity was localized in small granules, evenly dispersed throughout the entire nuclear envelop, and was observed as earlier as at 48 h. Cytoplasmic immunostaining for T?4 in HepG2 cells under starvation appeared significantly lower at 48 h and decreased progressively at 72 and at 84 h. At these time points, the decrease in cytoplasmic staining was associated with a progressive increase in nuclear reactivity, suggesting a possible translocation of the peptide from the cytoplasm to the nuclear membrane. The normal immunocytochemical pattern was restored when culture cells submitted to starvation for 84 h received a new complete medium for 48 h. Mass spectrometry analysis, performed on the nuclear and cytosolic fractions of HepG2 growing with and without serum, showed that T?4 was detectable only in the cytosolic and not in the intranuclear fraction. These data suggest that T?4 is able to translocate from different cytoplasmic domains to the nuclear membrane and back, based on different stress conditions within the cell. The punctuate pattern of nuclear T?4 immunostaining associated with T?4 absence in the nucleoplasm suggest that this peptide might be localized in the nuclear pores, where it could regulate the pore permeability.

Dataset Information

Thymosin Beta 4 Is Involved in the Development of Electroacupuncture Tolerance.

Publications

Thymosin Beta 4 Is Involved in the Development of Electroacupuncture Tolerance.

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