Comparison and effect of moxibustion and acupuncture on Nogo/NgR signaling pathway in rats with cerebral ischemia/reperfusion injury.

Background and aim: In China, acupuncture and moxibustion have been used effectively to treat various diseases for thousands of years. However, the evidence for a difference in the efficacies of moxibustion and acupuncture in cerebral infarction treatment is scarce. We aimed to compare the effects of acupuncture and moxibustion treatment on the Nogo/NgR signaling pathway in rats with cerebral ischemia/reperfusion (I/R) injury. Experimental procedure: Eighty male SD rats were randomly divided into five groups, based on treatment received: sham surgery (sham group), middle cerebral artery occlusion (MCAO, MCAO group), MCAO and NEP(1-40) inhibitor injection (MCAO + block group), MCAO and moxibustion (MCAO + moxi group), and MCAO and minimal acupuncture (MCAO + MA group). Neurological status was evaluated before treatment, and cerebral infarction volume (IV) and neurological function; Nogo-A, NgR, p75NTR, and LINGO-1 expressions; and NgR and LINGO-1 co-expression were assessed after treatment. Results and conclusion: After treatment, barring Nogo-A mRNA and protein expression in the MCAO + block group, the Longa score and IV significantly decreased; Nogo-A, NgR, p75NTR, and LINGO-1 mRNA and protein expressions as well as NgR and LINGO-1 co-expression significantly decreased in cerebral tissues; whereas the BWT score increased (P < 0.01) in the MCAO + moxi group, compared with the MCAO group. Except for NgR and LINGO-1 protein expressions, there were no significant differences in the abovementioned parameters between rats that underwent acupuncture and moxibustion. Acupuncture and moxibustion have similar effects on Nogo/NgR signaling pathway inhibition after cerebral infarction.

[Electroacupuncture improves limb motor function by down-regulating cytosolic phospholi-pase A2 and reducing apoptosis of nerve cells in rats with spinal cord injury].

OBJECTIVE: To observe the effect of electroacupuncture（EA） on the expression of cytosolic phospholipase A2 （cPLA2） and apoptosis of nerve cells in rats with spinal cord injury （SCI）, so as to explore its mechanisms underlying improvement of SCI. METHODS: Seventy-two female SD rats were randomly divided into model, EA, antagonist and EA+antagonist groups, with 18 rats in each group and other 18 rats were used as the sham operation （sham） group. The SCI model was established by referring to modified Allen's method with a weight impactor. The hindlimb motor function was assessed by using Basso-Beattie-Bresnahan （BBB） score. Rats of the EA group were subjected to EA stimulation at "Dazhui"（GV14）, "Yaoyangguan"（GV3）, bilateral "Ciliao"（BL32） and "Zusanli"（ST36） for 20 min, once a day for 14 days. Rats of the antagonist group received intravenous injection followed by intraperitoneal injection of arachidonyl trifluoromethyl ketone （AACOCF3, antagonist of cPLA2）, once every other day. Rats of the EA+antagonist group received EA treatment combined with antagonist injection. After the treatment, the rats were sacrificed and the spinal cord tissue was collected for detecting the protein expression of cPLA2, p-cPLA2, Bcl-2, Bax and Caspase-3 by Western blot, and the mRNA expression of cPLA2, Bcl-2, Bax and Caspase-3 using qRT-PCR. The morphological changes of the spinal cord were detected by Nissl staining. RESULTS: In comparison with the sham group, the BBB score, expression of Bcl-2 protein and mRNA were significantly down-regulated （P<0.01）, whereas the expression levels of Bax, Caspase-3 and p-cPLA2 proteins and mRNAs were considerably up-regulated in the model group （P<0.01）. Compared with the model group, the BBB score, expression levels of Bcl-2 protein and mRNA were significantly up-regulated （P<0.01, P<0.05）, while the expression levels of Bax, Caspase-3 and p-cPLA2 proteins in the EA, antagonist and EA+antagonist groups, Bax and cPLA2 mRNAs in both antagonist and EA+antagonist groups, and Caspase-3 mRNA in the EA+antagonist group were obviously down-regulated （P<0.01, P<0.05）. The effect of EA+antagonist was significantly superior to EA in increasing BBB score and in lowering expression of Bax and cPLA2 mRNAs （P<0.01, P<0.05）. Nissl staining showed reduced number of nerve cells and Nissl bodies, and striped dark blue cells in the model group, which was milder in the EA and antagonist groups, particularly in the EA+antagonist group. CONCLUSION: EA may improve the limb motor function of SCI rats, which may be related to its functions in down-regulating the expression of p-cPLA2, Bax and Caspase-3 and up-regulating Bcl-2 to reduce the apoptosis of nerve cells in the regional spinal cord.

[Effect of moxibustion on Nogo/neurotrophin receptor signaling pathway in rats with cerebral ischemia/reperfusion injury].

OBJECTIVE: To observe the effect of moxibustion treatment on the expression of Nogo-A, Nogo receptor (NgR), neurotrophin receptor p75 (p75NTR) and leucine rich repeat and Ig domain containing 1 (Lingo-1) in brain tissue of rats with cerebral ischemia/reperfusion injury (CI/RI), so as to analyze its mechanism underlying improvement of CI/RI. METHODS: Male SD rats were randomly divided into sham operation group (16 rats), model group (17 rats), NEP1-40 (extracellular peptide residues 1-40, a blocker targeting NgR) group (model+blocker, 17 rats) and moxibustion group (model+moxibustion, 17 rats). The CI/RI model was established by occlusion of the left middle cerebral artery (MCAO). Moxibustion was applied to "Baihui"(GV20), right "Quchi"(LI11) and "Zusanli"(ST36) for 20 min, once a day for 14 days, with 2 days' rest after the top 7 days' intervention. For rats of the NEP1-40 group, 30 µL PBS containing 18 µg NEP 1-40 was injected into the epidural inferior vena (L5-S1) via a polyvinyl chloride conduit. The neurological deficit state in each group was evaluated by Longa's 5-point scale and Feeney's 7-point scale of beam walking test (BWT). The cerebral infarct volume was assessed by 2,3,5-triphenyltetrazole chloride staining. The brain tissue between the central anterior and posterior sulcus was taken for observing the expression of NgR and Lingo-1 by fluorescence double-label method, and for determining the expression levels of Nogo-A, NgR, p75NTR and Lingo-1 mRNAs and proteins by real-time quantitative PCR and Western blot, respectively. RESULTS: After modeling, the Longa's score, infarct volu-me percent, expression levels of Nogo-A, NgR, Lingo-1 and p75NTR mRNAs and proteins were significantly increased (P<0.01) and BWT score was obviously decreased (P<0.01) in the model group relevant to the sham operation group. In comparison with the model group, the increase of Longa's score, infarct volume percentage, expression levels of Nogo-A, NgR, Lingo-1 and p75NTR mRNAs and proteins and decrease of BWT score in NEP1-40 and moxibustion groups were reversed (P<0.01) except Nogo-A protein in the NEP1-40 group. The effect of moxibustion was significantly superior to that of blocker NEP1-40 in redu-cing the infarct volume percentage, and down-regulating the expression of Nogo-A mRNA and protein, p75NTR mRNA and protein, NgR and Lingo-1 proteins (P<0.01, P<0.05). CONCLUSION: Moxibustion, similar to blocker NEP1-40 of NgR, can improve neurological dysfunction in CI/RI rats, which may be related to its functions in reducing cerebral infarction and down-regulating the activity of Nogo/neurotrophin receptor signaling pathway.

The mechanism of electroacupuncture for treating spinal cord injury rats by mediating Rho/Rho-associated kinase signaling pathway.

Objective: To determine the changes of gene and protein expression through Rho/ROCK signaling pathway in EA treated spinal cord injury (SCI) rats and to unveil the possible underlying mechanism.Design: Animal study.Setting: Affiliated Hospital of Jiangxi University of Traditional Chinese Medicine.Participants: Eighty Male Sprague Dawley rats.Interventions: Electroacupuncture at Yaoyangguan (GV3), Dazhui (GV14), Zusanli (ST36) and Ciliao (BL32) and/or blocking agent Y27632 treatment.Outcome Measures: Protein expression was detected by ELISA and Western blotting, mRNA expression was detected by quantitative PCR and in situ hybridization. Morphological changes in spinal cord were evaluated by HE-staining and Nissl staining. Hindlimb motor function in the rats was evaluated by Basso-Beattie-Bresnahan (BBB) assessment methods.Results: Compared with injured rats in SCI group, EA, blocking agent Y27632 and EA + blocking agent Y27632 treatment had significantly reduced mRNA and protein expression levels of RhoA and ROCKII, decreased p-MLC protein expression and p-MLC/MLC ratio, suppressed cPLA2 activity and PGE2 level, improved spinal cord tissue morphology and BBB score of lower limb movement function at 7 days and at 14 days (P < 0.01 or <0.05).Conclusion: Similar to the blocking agent Y27632, EA may have a notable inhibitory effect on the Rho/ROCK signaling pathway after SCI, therefore reducing the inhibition of axonal growth and inflammatory reaction may be a key mechanism of EA treatment for SCI.

Electroacupuncture Promoting Axonal Regeneration in Spinal Cord Injury Rats via Suppression of Nogo/NgR and Rho/ROCK Signaling Pathway.

PURPOSE: To observe the changes of Nogo/NgR and Rho/ROCK signaling pathway-related gene and protein expression in rats with spinal cord injury (SCI) treated with electroacupuncture (EA) and to further investigate the possible mechanism of EA for treating SCI. METHODS: Allen's method was used to create the SCI rat model. Sixty-four model rats were further subdivided into four subgroups, namely, the SCI model group (SCI), EA treatment group (EA), blocking agent Y27632 treatment group (Y27632) and EA+blocking agent Y27632 treatment group (EA+Y), according to the treatment received. The rats were subjected to EA and/or blocking agent Y27632 treatment. After 14 days, injured spinal cord tissue was extracted for analysis. The mRNA and protein expression levels were determined by real-time fluorescence quantitative PCR and Western blotting, respectively. Cell apoptosis changes in the spinal cord were evaluated by in situ hybridization. Hindlimb motor function in the rats was evaluated by Basso-Beattie-Bresnahan assessment methods. RESULTS: Except for RhoA protein expression, compared with the SCI model group, EA, blocking agent Y27632 and EA+blocking agent Y27632 treatment groups had significantly reduced mRNA and protein expression of Nogo-A, NgR, LINGO-1, RhoA and ROCK II in spinal cord tissues, increased mRNA and protein expression of MLCP, decreased p-MYPT1 protein expression and p-MYPT1/MYPT1 ratio, and caspase3 expression, and improved lower limb movement function after treatment for 14 days (P<0.01 or <0.05). The combination of EA and the blocking agent Y27632 was superior to EA or blocking agent Y27632 treatment alone (P < 0.01 or <0.05). CONCLUSION: EA may have an obvious inhibitory effect on the Nogo/NgR and Rho/ROCK signaling pathway after SCI, thereby reducing the inhibition of axonal growth, which may be a key mechanism of EA treatment for SCI.

Effect of electroacupuncture on the mRNA and protein expression of Rho-A and Rho-associated kinase II in spinal cord injury rats.

Electroacupuncture is beneficial for the recovery of spinal cord injury, but the underlying mechanism is unclear. The Rho/Rho-associated kinase (ROCK) signaling pathway regulates the actin cytoskeleton by controlling the adhesive and migratory behaviors of cells that could inhibit neurite regrowth after neural injury and consequently hinder the recovery from spinal cord injury. Therefore, we hypothesized electroacupuncture could affect the Rho/ROCK signaling pathway to promote the recovery of spinal cord injury. In our experiments, the spinal cord injury in adult Sprague-Dawley rats was caused by an impact device. Those rats were subjected to electroacupuncture at Yaoyangguan (GV3), Dazhui (GV14), Zusanli (ST36) and Ciliao (BL32) and/or monosialoganglioside treatment. Behavioral scores revealed that the hindlimb motor functions improved with those treatments. Real-time quantitative polymerase chain reaction, fluorescence in situ hybridization and western blot assay showed that electroacupuncture suppressed the mRNA and protein expression of Rho-A and Rho-associated kinase II (ROCKII) of injured spinal cord. Although monosialoganglioside promoted the recovery of hindlimb motor function, monosialoganglioside did not affect the expression of Rho-A and ROCKII. However, electroacupuncture combined with monosialoganglioside did not further improve the motor function or suppress the expression of Rho-A and ROCKII. Our data suggested that the electroacupuncture could specifically inhibit the activation of the Rho/ROCK signaling pathway thus partially contributing to the repair of injured spinal cord. Monosialoganglioside could promote the motor function but did not suppress expression of RhoA and ROCKII. There was no synergistic effect of electroacupuncture combined with monosialoganglioside.

Gender differences in brain regional homogeneity of healthy subjects after normal sleep and after sleep deprivation: a resting-state fMRI study.

OBJECTIVE: To explore the gender differences of brain regional homogeneity (ReHo) in healthy subjects during the resting-state, after normal sleep, and after sleep deprivation (SD) using functional magnetic resonance imaging (fMRI) and the ReHo method. METHODS: Sixteen healthy subjects (eight males and eight females) each underwent the resting-state fMRI exams twice, i.e., once after normal sleep and again after 24h's SD. According to the gender and sleep, 16 subjects were all measured twice and divided into four groups: the male control group (MC), female control group (FC), male SD group (MSD), and female SD group (FSD). The ReHo method was used to calculate and analyze the data, SPM5 software was used to perform a two-sample T-test and a two-pair T-test with a P value <0.001, and cluster volume ≥ 270 mm(3) was used to determine statistical significance. RESULTS: Compared with the MC, the MSD showed significantly higher ReHo in the right paracentral lobule (BA3/6), but in no obviously lower regions. Compared with the FC, the FSD showed significantly higher ReHo in bilateral parietal lobes (BA2/3), bilateral vision-related regions of occipital lobes (BA17/18/19), right frontal lobe (BA4/6), and lower ReHo in the right frontal lobe. Compared with the FC, the MC showed significantly higher ReHo in the left occipital lobe (BA18/19), and left temporal lobe (BA21), left frontal lobe, and lower ReHo in the right insula and in the left parietal lobe. Compared with the FSD, the MSD showed significantly higher ReHo in the left cerebellum posterior lobe (uvula/declive of vermis), left parietal lobe, and bilateral frontal lobes, and lower ReHo in the right occipital lobe (BA17) and right frontal lobe (BA4). CONCLUSIONS: The differences of brain activity in the resting state can be widely found not only between the control and SD group in a same gender group, but also between the male group and female group. Thus, we should take the gender differences into consideration in future fMRI studies, especially the treatment of brain-related diseases (e.g., depression).

[Evaluation of the post-effect of acupuncture at Sanyinjiao (SP 6) under sleep deprivation by resting-state amplitude of low-frequency fluctuation: a fMRI study].

OBJECTIVE: To discuss the central modulating mechanism of Sanyinjiao (SP 6) and the amplitude of low-frequency fluctuation (ALFF) differences of the functional brain regions between SP 6 and sham acupoint. METHODS: Sixteen volunteers accepted right Sanyinjiao (SP 6) (SP 6 group) or right sham (sham group) acupuncture for two times after 24 h sleep deprivation (SD), there was two weeks between two acupunctures. The needle was retained for 4 minutes and the brain fMRI scanning was performed by 3.0 TMR every time. The ALFF consequent data was processed by REST. One-sample t-test and two-pair t-test were performed by SPM5. RESULTS: SP 6 group mainly caused ALFF change in inferior parietal lobule, posterior cingulated while sham group in precuneus, posterior cingulated. SP 6 group increased ALFF in left fusiform temporal gyrus, medial frontal gyrus with no decreased regions compared with sham group. CONCLUSIONS: Retaining needle dose influence the activity of the brain region during resting-state in both group, especially the mood-related regions. And its mechanism is probably that the relevant effect is carried out by regulating function to target organs of complex brain network comprised of relevant functional center and related brain region. There are differences between SP 6 and Sham acupoint.

[Brief analysis on scientifity of the international scalp acupuncture].

By comparing and analyzing the similarity among the international scalp acupuncture, JIAO's scalp acupuncture and ZHU's scalp acupuncture, it is believed that the international scalp acupuncture is a combination of traditional acupuncture and other scalp acupuncture on the basis of channel theory of TCM, and this combination is worthy of discussion. However JIAO's scalp acupuncture lacks scientific evidence. So, the international scalp acupuncture following most contents of JIAO's scalp acupuncture is worthy of discussion.

[Preliminary study on force feedback of acupuncture in virtual reality based on the visible human].

This paper discusses the application of virtual reality technology in the 3-D visible human body and acupuncture research. Based on the 3-D visible human fused with the localization information and hierarchy of acupoints, the paper analyzes the force against the needle and haptic rendering during the needle manipulation according to the physical properties of different tissues. A haptic model is constructed to demonstrate the force behaviors during acupuncture, and the force will be produced and passed to the manipulator by a force feedback device. It enriches the contents of 3-D visible human project, provides a dynamic simulation instrument for acupuncture teaching, and supplies a platform for acupuncture research.

[Three-D visualization study on the acupoint of Jianliao (TE 14) based on the operational platform of Voxel-man].

OBJECTIVE: To discuss the problem difficult to be presented for nerves and blood vessels in the 3D reconstruction of acupoints through studying 3D reconstruction of Jianliao (TE 14). METHODS: Muscles and other tissues were segmented by applying cointeractive segmentation method based on chromatic characteristic space; the nerve and blood vessels can be reconstructed by establishing mathematics model, and the needle-inserting animation of Jianliao (TE 14) could be obtained by running script file. RESULTS: During the course of 3D needle-inserting animation of Jianliao (TE 14), the spatial location between all kinds of tissues including the nerve and blood vessels and the needle body could be systematically observed. CONCLUSION: The difficulty of presenting the nerve and blood vessels in the study on 3D reconstruction of acupoints can be resolved by establishing mathematics model. The generation of 3D animation for acupoints with the nerve and blood vessels is benefit to acupuncture teaching and study on the essence of acupoints and channels.