Electroacupuncture improves post-traumatic stress disorder in rats by alleviating hippocampal mitochondrial injury via regulating Bcl-2/Bax/caspase-3 signaling

doi:10.12122/j.issn.1673-4254.2025.11.10

Abstract

Abstract:

Objective To investigate the mechanism underlying the therapeutic effect of electroacupuncture (EA) on post-traumatic stress disorder (PTSD) in rats. Methods Forty male SD rats were randomized equally into blank control group, PTSD model group, sham-acupuncture group, paroxetine group, and EA group. In the latter 3 groups, the rat models of PTSD, induced by continuous single-prolonged stress and plantar electrical stimulation, were treated with EA at GV20, GV24, BL18 and BL23 acupoints for 15 min (5 times a week for 3 weeks), sham-acupuncture without electrical stimulation, or gavage with paroxetine suspension on the same schedule. Behavioral changes of the rats were evaluated using open field test (OFT) and elevated plus maze (EPM) test. Hippocampal pathologies and neuronal changes were examined with HE and Nissl staining, and mitochondrial ultrastructure was examined using electron microscopy. The mRNA and protein expression levels of Bcl-2, Bax, and caspase-3 were detected by RT-qPCR and immunofluorescence staining. Results The rat models of PTSD showed significantly reduced total distance traveled in OFT and distance and time spent in the open arms of the EPM, with decreased hippocampal neurons, obvious neuronal and mitochondrial pathologies, decreased hippocampal expression of Bcl-2, and increased Bax and caspase-3 expressions. Treatments with paroxetine and EA both significantly improved behavioral changes of the rat models, increased the number of Nissl-stained neurons, obviously alleviated pathologies in the hippocampal neurons and mitochondrial ultrastructure, increased hippocampal Bcl-2 expression, and lowered caspase-3 expressions. Paroxetine showed significantly better effect than EA for improving performance of the rats in EPM test, whereas sham-acupuncture did not produce any significant improvement. Conclusion EA alleviates PTSD in rats possibly by upregulating Bcl-2 and downregulating Bax and caspase-3, thereby ameliorating hippocampal mitochondrial damage.

Key words: post-traumatic stress disorder, electroacupuncture, mitochondria, neurons, caspase-3, anxiety disorder

Dandan MA, Jie CHENG, Hong ZHANG, Guang LIU, Kai² SONG. Electroacupuncture improves post-traumatic stress disorder in rats by alleviating hippocampal mitochondrial injury via regulating Bcl-2/Bax/caspase-3 signaling[J]. Journal of Southern Medical University, 2025, 45(11): 2375-2384.

Figures/Tables 8

Tab.1 Primer sequences for RT-qPCR and product lengths

Target gene	Primer sequences	Product Length (bp)
Bcl-2	CTGGTGGACAACATCGCTCT(F) GCATGCTGGGGCCATATAGT(R)	115
Bax	CAACATGGAGCTGCAGAGGA(F) GGAAAGGAGGCCATCCCAG (R)	274
Caspase-3	GAGCTTGGAACGCGAAGAAA(F) TTGCGAGCTGACATTCCAGT (R)	221
GAPDH	CCTCGTCCCGTAGACAAAATG (F) TGAGGTCAATGAAGGGGTCGT (R)	133

Fig.1 Anxiety-like behaviors of the rats in the 5 groups (Mean±SD, n=8). A: Motion trajectory in open field test. B: Statistics of total distance traveled. ****P<0.0001 vs Control group; ####P<0.0001 vs Model group.

Fig.2 Anxiety-like behaviors of the rats in the 5 in elevated plus maze test (Mean±SD, n=8). A: Representative movement trajectories in the elevated plus maze test. B: Statistics of open arm total distance. C: Open arm total time. *P<0.05, ****P<0.0001 vs Control group; ##P<0.01, ####P<0.0001 vs Model group.

Fig.3 Pathological and morphological characteristics of hippocampal neurons of the rats in the 5 rats (HE staining, scale bar=20 μm). Yellow arrows: Normal neurons; Black arrows: Shrunken neurons; Green arrows: Vacuoles; Red arrows: Perivascular edema.

Fig.4 Comparison of neuronal structure in the hippocampal region of the rats among the 5 groups (Nissl staining, scale bar=50 μm). A: Representative photomicrographs.B: Neuronal cell count (n=3).****P<0.0001 vs Control group; #P<0.05 vs Model group.

Fig.5 Comparison of mitochondrial morphology in the hippocampal neurons among the 5 groups (scale bar=500 nm). Red arrows indicate mitochondria.

Fig.6 Expression levels of Bcl-2 (A), Bax (B), and caspase-3 (C) mRNA in the hippocampus of the rats in the 5 groups (Mean±SD, n=3). ****P<0.0001 vs Control group; ###P<0.001, ####P<0.0001 vs Model group.

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