Electroacupuncture Improves Cerebral Ischemic Injury by Enhancing the EPO-JAK2-STAT5 Pathway in Rats.

OBJECTIVE: Clinically, electroacupuncture (EA) improves cerebral ischemic injury, but its mechanism remains unknown. The aim of this study was to confirm the protective effects of EA on focal cerebral ischemia (FCI)-induced injury and the possible mechanism. METHODS: Sprague-Dawley (SD) rats served as the FCI model and were divided into the sham, model, EA, AG490 and EA+AG490 groups. Rats in the EA and EA+AG490 groups were acupunctured at the Baihui (GV20) and Dazhui (GV14) acupoints, and those in the AG490 and EA+AG490 groups were administered an intracerebroventricular injection of AG490 (a Janus-tyrosine kinase-2 (JAK-2) phosphorylation inhibitor). Neurological deficits and morphological changes in the ischemic cortex were observed through neurological deficit scoring and HE staining, respectively, and neuronal apoptosis was examined using the TUNEL assay. Transmission electron microscopy was used to observe neuronal ultrastructure, and HIF-1α, erythropoietin (EPO), phosphorylated (p)-JAK2, p-STAT5, HSP70, Bax and Bcl-2 expression was measured by RT-PCR and immunohistochemistry. RESULTS: FCI model rats showed obvious neurological deficits and neuronal apoptosis compared with sham rats. EA alleviated FCI-induced neurological deficits, improved neuronal ultrastructure, reduced neuronal apoptosis, and induced HIF-1α, EPO, p-JAK2, p-STAT5, HSP70 and Bcl-2 expression in a time-dependent manner. In contrast, AG490 treatment impaired the effects of EA on neurological deficits, neuronal apoptosis and HIF-1α, EPO, p-JAK2, p-STAT5, HSP70, Bax and Bcl-2 expression. CONCLUSION: EA at GV20 and GV14 could improve neurological deficits and reduce neuronal apoptosis, thereby improving FCI-induced injury, which may be related to enhancing the EPO-JAK2-STAT5 pathway.

Electroacupuncture improves cognitive deficits and insulin resistance in an OLETF rat model of Al/D-gal induced aging model via the PI3K/Akt signaling pathway.

To investigate the effect of electroacupuncture (EA) on cognitive function and insulin resistance (IR) in an Al/D-gal-induced aging model for Alzheimer's disease (AD) using Ostuka Long-Evans Tokushima Fatty (OLETF) rats. The Al/D-gal-OLETF rats for AD were randomly divided into the EA and non-EA groups. Cognitive function was assessed using the Morris water maze (MWM). The morphology of the hippocampal neurons was observed using hematoxylin & eosin (H&E) staining. Aß and total Tau in the hippocampus and cerebrospinal fluid (CSF) were detected using western blotting (WB) and enzyme-linked immunosorbent assay (ELISA). Fasting blood glucose (FPG) was determined using the glucose oxidase method. Plasma fasting insulin (FINS), serum C-peptide (C-P), and CSF insulin were detected using ELISA. The expression of the genes and proteins in the PI3K signaling pathway was detected using quantitative real-time PCR and WB. After EA intervention, the hippocampal Aß and total Tau protein levels, body weight, FPG, FINS, and C-P were significantly decreased. The MWM showed that the percentage of time in the target quadrant of the EA group was elevated in the probe test. The protein levels of p-IRS1, p-IRS2, IDE, and p-GSK3ß were significantly increased, while p-PI3K-p85α and p-Akt were decreased. In conclusion, EA improves cognitive function and insulin resistance in rat models of AD. The PI3K/Akt signaling pathway is involved in those changes.

Acupuncture Decreases NF-κB p65, miR-155, and miR-21 and Increases miR-146a Expression in Chronic Atrophic Gastritis Rats.

Acupuncture has been used to treat chronic atrophic gastritis (CAG) in traditional Chinese medicine (TCM) for centuries. In this study, we evaluated the effect of acupuncture at Zusanli (ST36), Zhongwan (CV12), and Pishu (BL20) acupoints on weight changes of rats, histological changes of gastric glands, and expressions changes of nuclear factor-kappa B (NF-κB) p65, microRNA- (miR-) 155, miR-21, and miR-146a in CAG rats induced by N-methyl-N'-nitro-N-nitrosoguanidine (MNNG) combined with irregular diet. Consequently, we found that acupuncture treatment elevated body weight of rats significantly when compared to the model group. By observing histological changes, we found that the acupuncture group showed better improvement of gastric mucosa injury than the model group. Our results also demonstrated upregulation of NF-κB p65, miR-155, and miR-21 in gastric tissue of CAG rats and a positive correlation between miR-155 and miR-21. Relatively, expression of miR-146a was downregulated and negative correlation relationships between miR-146a and miR-155/miR-21 in CAG rats were observed. Additionally, expressions of NF-κB p65, miR-155, and miR-21 were downregulated and miR-146a was upregulated after acupuncture treatment. Taken together, our data imply that acupuncture can downregulate NF-κB p65, miR-155, and miR-21 and upregulate miR-146a expression in CAG rats. NF-κB p65, miR-155, miR-21, and miR-146a may play important roles in therapeutic effect of acupuncture in treating CAG.

Neuroprotective effects of electroacupuncture on hypoxic-ischemic encephalopathy in newborn rats are associated with increased expression of GDNF-RET and protein kinase B.

OBJECTIVE: To explore the neuroprotective effects of electroacupuncture (EA) on hypoxic-ischemic encephalopathy (HIE) and to further investigate the role of glial cell line-derived neurotrophic factor (GDNF) family receptor member RET (rearranged during transfection) and its key downstream phosphatidylinositol 3 kinase (PI-3K)/protein kinase B (Akt) pathway in the process. METHODS: A total of 220 seven-day-old SD rats (of either sex, from 22 broods) were randomly divided into two groups, one (30 rats) for sham-surgery group and the other (190 rats) for HIE model group. The HIE model was established using the left common carotid artery ligation method in combination with hypoxic treatment. The successfully established rats were randomly divided into five groups, including control model group, EA group, sham-EA group, antagonist group and antagonist plus electroacupuncture group, with 35 rats in each group. Baihui (GV 20), Dazhui (GV 14), Quchi (LI 11) and Yongquan (KI 1) acupoints were chosen for acupuncture. EA was performed at Baihui and Quchi for 10 min once a day for continuous 1, 3, 7 and 21 days, respectively. The rats were then killed after the operation and injured cerebral cortex was taken for the measurement of neurologic damage by hematoxylin-eosin (HE) staining and the degenerative changes of cortical ultrastructure by transmission electron microscopy. RET mRNA level and Akt protein level were detected by real-time reverse-transcription polymerase chain reaction (RT-PCR) and western blot analysis, respectively. RESULTS: EA could ameliorate neurologic damage of the first somatic sensory area (S1Tr) and alleviate the degenerative changes of ultrastructure of cortical neurons in rats subjected to HIE. And the longer acupuncture treatment lasted, the better its therapeutic effect would be. This was accompanied by gradually increased expression of GDNF family receptor RET at the mRNA level and its downstream signaling Akt at the protein level in the ischemic cortex. CONCLUSION: EA has neuroprotective effects on HIE and could be a potential therapeutic strategy for HIE in the neonate. Activation of RET/Akt signaling pathway might be involved in this process.

[Effect of electroacupuncture on expression of signal transducer and activator of transcription 3 in focal ischemic cerebral tissue in cerebral ischemia rats].

OBJECTIVE: To observe the effect of electroacupuncture (EA) intervention on expression of signal transducer and activator of transcription 3 (STAT 3) in the focal ischemic cerebral tissue, so as to study its mechanism underlying improving ischemic stroke. METHODS: A total of 150 SD rats were randomized into sham operation (control) group, cerebral ischemia (CI) model (model) group and EA group which were further randomly divided into 2 hour (2 h), 1 day (1 d), 3 d, 1 week (1 W) and 3 W subgroups (n = 6/subgroup for immunohistochemistry, n = 4/subgroup for Western blot). CI model was established by occlusion of the middle cerebral artery with electro-coagulation method. EA (3 Hz/20 Hz, 2-3 V) was applied to "Baihui" (GV 20) and "Dazhui "(GV 14) for 30 min. The expression of cerebral STAT 3 was detected by immunofluorescence histochemistry and laser-confocal microscopy, and Western blot, separately. RESULTS: Compared with the control group, cerebral STAT 3 immunofluorescence intensity values at the time-points of 2 h, 1 d, 3 d and 1 W, STAT 3 protein expression levels at the time-points of 2 h, 1 d and 3 d in the model group were increased significantly (P < 0.001, P < 0.05). After acupuncture intervention, cerebral STAT 3 immunofluorescence intensity values at the time-points of 1 d, 3 d, 1 W and 3 W, STAT 3 protein expression levels at the time-points of 1 d, 3 d and 3 W in the EA group were down-regulated considerably (P < 0.001, P < 0.01, P < 0.05). No significant differences were found between the control and model groups in STAT 3 immunofluorescence intensity at 3 W, and in STAT 3 protein expression levels at 1 W and 3 W, and between the EA and model groups in STAT 3 immunofluorescence intensity at 2 h, and in STAT 3 protein expression at 2 h, 3 d and 1 W (P > 0.05). CONCLUSION: EA therapy can down-regulate the expression level of STAT 3 protein in the regional ischemic cerebral tissue in cerebral ischemia rats, which may contribute to its efficacy in the treatment of acute and chronic ischemic stroke.

Electroacupuncture effect on neurological behavior and tyrosine kinase-JAK 2 in rats with focal cerebral ischemia.

OBJECTIVE: Electroacupuncture effect on neurological behavior and the expression of tyrosine kinase Janus kinase 2 (JAK 2) of ischemic cortex in rats with the focal cerebral ischemia were investigated in this study. METHODS: The model of focal cerebral ischemia was established by the heat-coagulation induced the occlusion of the middle cerebral artery. The electro-acupuncture was applied on Baihui (GV 20) and Dazhui (GV 14), and AG490 was applied by intracerebroventricular infusion. The expressions of JAK2 mRNA and phospharylated JAK2 (p-JAK2) in the ischemic cortex were observed by in situ hybridization and western blotting. RESULTS: The expressions of JAK2 mRNA and p-JAK2 were rarely found in sham surgery group. In model group, the expression of JAK2 mRNA and JAK2 phosphorylation had increased. After 1 day of cerebral ischemia, the expression had reached its peak. After cerebral ischemia, the expressions of JAK2 mRNA and p-JAK2 were consistent with the neurological deficit score. Electroacupuncture treatment and AG490 intervention were able to improve the neurological deficit score after cerebral ischemia, and down-regulate the expressions of JAK2 mRNA and JAK2 phosphorylation. CONCLUSION: After cerebral ischemia, the excessive expressions of JAK2 and the JAK2 phosphorylation would be one of mechanisms by which the brain injury got worse. The therapy of electro-acupuncture could reduce the expression of JAK2, and inhibit JAK2 phosphorylated activation, so as to block the abnormal activation of signal transduction pathway which was induced by JAK2.

Electroacupuncture effects on cortical neurons, as well as Janus kinase 2-signal transducer and activator of transcription 3 signal transduction pathway, in a rat model of cerebral ischemia.

The present study established a model of focal cerebral ischemia through heat-coagulation induced occlusion of the middle cerebral artery. Following electroacupuncture at Baihui (GV20) and Dazhui (GV14), or intracerebroventricular infusion of AG490, a Janus kinase 2 phosphorylation inhibitor, the amount of necrotic or degenerated neurons in the ischemic cerebral cortex decreased, neuronal swelling was ameliorated, and expression of phosphorylated Janus kinase 2 and signal transducer and activator of transcription 3 decreased. Results confirmed that electroacupuncture promoted neuronal repair in the cerebral cortex by reducing expression of phosphorylated Janus kinase 2 and signal transducer and activator of transcription 3, as well as weakening the phosphorylated activation, thereby blocking abnormal activation of the Janus kinase 2- signal transducer and activator of transcription 3 signal transduction pathway.