A preliminary investigation of the regulatory effect of acupuncture on hippocampal circHDAC2/miR-3065/SLC30A3 axis in CIRI rats. / å¤§é¼ æµ·é©¬ç»„ç»‡circHDAC2/miR-3065/SLC30A3è½´å‚ä¸Žé’ˆåˆºè°ƒèŠ‚è„‘ç¼ºè¡€å†çŒæ³¨æŸä¼¤çš„åˆæ­¥ç ”ç©¶.

OBJECTIVES: To explore the mechanisms of acupuncture against cerebral ischemia/reperfusion injury (CIRI) through observing the expression of circular RNA HDAC2 (circHDAC2) in the hippocampus of rats. METHODS: SD rats were randomly divided into sham-operation, model and acupuncture groups, with 13 rats in each group. The rat model of CIRI was established by middle cerebral artery occlusion. In the acupuncture group, acupuncture was delivvered at "Dazhui" (GV14), "Shuigou" (GV26) and "Baihui" (GV20), and the needles were retained for 30 min each time and acupuncture was conducted once every 12 h for a total of 7 sessions. Before and after intervention, using modified Garcia scale, the neurological function of the rats were evaluated, and TTC staining was employed to determine the cerebral infarct area. Gene chip technology was used to screen the circRNAs with differential expressions in the ischemic hippocampus, and the circRNAs with co-differential expression (co-DE circRNAs) in the model group/sham-operation group, and the acupuncture group/model group separately. Among those circRNAs, the core circRNAs were screened according to P value, fold change (FC) and gene ontology (GO) analysis；and their expressions in the ischemic hippocampus were determined using quantitative real-time PCR (qPCR). Based on the verification results, a competing endogenous RNA (ceRNA) prediction network was constructed. The expression levels of microRNA (miRNA) and mRNA with high node centrality in the prediction network were validated by qPCR. RESULTS: Before intervention, compared with the sham-operation group, the modified Garcia score of each modeling group decreased (P<0.01). After intervention, the modified Garcia score was reduced and the cerebral infarct area ratio increased (P<0.01) in the model group when compared with the sham-operation group. In the acupuncture group, the modified Garcia score was higher and the cerebral infarct area ratio lower (P<0.01) than those of the model group. The microarray results of gene chip showed that 16 co-DE circRNAs were down-regulated in the model group and up-regulated in the acupuncture group, and 7 co-DE circRNAs up-regulated in the model group and down-regulated in the acupuncture group. The core circHDAC2 and circNTRK2 were screened according to P value, FC and the enrichment number of GO entries. QPCR results showed that, compared with the sham-operation group, the expression of circHDAC2 and circNTRK2 of the ischemic hippocampal tissue was down-regulated in the model group (P<0.01)；and that of circHDAC2 and circNTRK2 up-regulated in the acupuncture group when compared with the model group (P<0.01). The relevant ceRNA regulatory network was constructed for circHDAC2 and the prediction results showed that the regulatory networks contained 12 miRNAs and 31 mRNAs. Results of verifying miRNA with high network node centrality and mRNA relevant with nerve regulation showed that, when compared with the sham-operation group, the expression levels of miR-29a, miR-29b and the solute carrier family 30 member 3 (SLC30A3) mRNA in the ischemic hippocampus were down-regulated (P<0.01)；and those of miR-3065 and mercaptopyruvate sulfurtransferase (MPST) up-regulated (P<0.01) in the model group. Compared with the model group, the expressions of miR-29a, miR-29b and SLC30A3 mRNA in the ischemic hippocampus were up-regulated (P<0.01, P<0.05), while that of miR-3065 down-regulated (P<0.05) in the acupuncture group. CONCLUSIONS: Acupuncture significantly improves the neurological function and reduces the cerebral infarct area in CIRI rats, which may be related to the regulation of hippocampal circHDAC2/miR-3065/SLC30A3 axis.

[Acupuncture regulates autophagy in hippocampal neurons of cerebral ischemia/reperfusion injured rats by activating type â ¢ PI3K/Beclin-1 pathway].

OBJECTIVE: To observe the effects of acupuncture on the expression of type Ⅲ phosphatidylinositol 3-hydroxykinase (PI3K) and Beclin-1 in hippocampus of rats with cerebral ischemia/reperfusion injury (CI/RI), so as to explore the mechanism of acupuncture in regulating type Ⅲ PI3K pathway to activate autophagy in the hippocampal neurons of CI/RI rats. METHODS: SD rats were randomly divided into sham operation group (n=11) and operation group. Then after successful modeling, rats in the operation group were randomly divided into model, acupuncture, model+3-MA and acupuncture+3-MA groups, with 11 rats in each group. The model of CI/RI was established by occlusion of the middle cerebral artery. Rats in the model+3-MA and acupuncture+3-MA groups were injected with 3-MA (400 nmol/ 5 µL) 5 µL into the lateral ventricle 30 min before reperfusion. Rats in the acupuncture and acupuncture+3-MA groups were punctured with filiform needles at "Dazhui" (GV14), "Shuigou" (GV26) and "Baihui" (GV20) and stimulated manually once every 15 min. The acupuncture intervention was conducted for 30 min each time, once every 12 h for a total of 7 times. The degree of neurological impairment was evaluated 2 h after reperfusion and after intervention by Garcia score. After intervention, the percentage of cerebral ischemic area was observed by TTC staining, the protein expression levels of type Ⅲ PI3K, Beclin-1, microtubule-associated protein 1 light chain 3B (LC3B), lysosome associated membrane protein 2 (Lamp2) and P62 in ischemic hippocampal tissue were detected by Western blot, the ultrastructure of neurons in ischemic hippocampus was observed by transmission electron microscopy (TEM). RESULTS: Compared with the sham operation group, the Garcia score was decreased (P<0.01), the percentage of cerebral ischemic area was increased (P<0.01), the expression levels of type Ⅲ PI3K, Beclin-1, LC3B-Ⅱ/Ⅰ, Lamp2 proteins were decreased (P<0.01), and the expression level of P62 protein was increased (P<0.01) in ischemic hippocampal tissue in the model group. Compared with the model group, the Garcia score was increased (P<0.01), the percentage of cerebral ischemic area was decreased (P<0.01), the expression levels of type Ⅲ PI3K, Beclin-1, LC3B-Ⅱ/Ⅰ, Lamp2 proteins were increased (P<0.01, P<0.05) and the expression level of P62 was decreased (P<0.01) in ischemic hippocampal tissue in the acupuncture group； the percentage of cerebral ischemic area was increased (P<0.05), the expressions of type Ⅲ PI3K and Beclin-1 were decreased (P<0.01) and the expression level of P62 protein was increased (P<0.05) in ischemic hippocampal tissue in the mo-del+3-MA group. Compared with the model +3-MA group, the Garcia score was increased (P<0.05), the percentage of cerebral ischemic area was decreased (P<0.01), the expression levels of type Ⅲ PI3K, Beclin-1, LC3B-Ⅱ/Ⅰ in ischemic hippo-campal tissue were increased (P<0.01, P<0.05) in the acupuncture+3-MA group. Compared with the acupuncture group, the Garcia score was decreased, the percentage of cerebral ischemic area was increased (P<0.05, P<0.01), the expression levels of type Ⅲ PI3K, Beclin-1, Lamp2 proteins were decreased (P<0.01, P<0.05) and P62 protein was increased (P<0.05) in ischemic hippocampal tissue in the acupuncture+3-MA group. The results of TEM showed that the edema of neurons was heavier, and few hypolysosomes existed in the model group; there was no obvious damage to neuronal structure, intracellular matrix was abundant, and a few lysosomes existed in the acupuncture group; the neuronal cells had mild edema and primary lysosomes were present in the acupuncture +3-MA group. CONCLUSION: Acupuncture can improve the symptoms of neurological impairment and reduce the percentage of cerebral ischemic area in rats with CI/RI. The mechanism may be related to regulating type Ⅲ PI3K/Beclin-1 pathway, up-regulating the expressions of autophagy related factors LC3B-Ⅱ and Lamp2, and down-regulating the expression of P62.

[Effect of acupuncture technique of Tiaoxin Tongdu on learning-memory ability and expressions of hippocampal VEGF and Ang-1 in rats with vascular dementia].

OBJECTIVE: To observe the effect of acupuncture technique of Tiaoxin Tongdu on learning-memory ability and expressions of hippocampal vascular endothelial growth factor (VEGF) and angiogenin-1 (Ang-1) in rats with vascular dementia (VD), and to explore the mechanism of acupuncture technique of Tiaoxin Tongdu for VD. METHODS: A total of 24 male SD rats were randomly divided into a sham operation group, a model group, a medication group and an acupuncture group after Morris water maze test, 6 rats in each group. VD model was established by permanent ligation of bilateral common carotid arteries in the model group, the medication group and the acupuncture group. Treatment was given on the next day after successful modeling. The rats in the acupuncture group were treated with acupuncture at "Baihui" (GV 20), "Shenting" (GV 24), "Shuigou" (GV 26), "Dazhui" (GV 14), "Fengfu" (GV 16), "Mingmen" (GV 4), "Neiguan" (PC 6), "Daling" (PC 7) and "Laogong" (PC 8) for 30 min; the rats in the medication group were treated with nimodipine solution (0.0625 g/kg) by gavage, once a day, for 2 weeks. Morris water maze test was used to detect the behavior of rats before modeling, 2 weeks after modeling and after intervention; after intervention, the expressions of VEGF and Ang-1 protein in hippocampus were detected by Western blot. RESULTS: Compared with the sham operation group, the average escape latency of rats in the model group was prolonged (P<0.01), and the times of crossing the original platform were reduced (P<0.01). Compared with the model group, the average escape latency of rats in the medication group and acupuncture group was significantly shortened (P<0.01), and the times of crossing the original platform were increased (P<0.01, P<0.05). Compared with the sham operation group, the expressions of VEGF and Ang-1 protein in hippocampus in the model group were increased (P<0.05, P<0.01). Compared with the model group, the expressions of VEGF and Ang-1 protein in the hippocampus in the medication group and acupuncture group were significantly increased (P<0.01, P<0.05). CONCLUSION: The acupuncture technique of Tiaoxin Tongdu can significantly improve the learning and memory ability of VD rats, and its mechanism may be related to up-regulating the expressions of VEGF and Ang-1 protein in hippocampus and inducing angiogenesis.

[Effect of Acupuncture on Neurological Function and Related Differentially-depressed Profiles in the Brain of Middle Cerebral Artery Occlusion Rats Using Antibody Chips Technique].

OBJECTIVE: To observe the effect of acupuncture on changes of neurological function and expression of proteins in the ischemic brain region in middle cerebral artery occlusion (MCAO) rats with protein chip technique, so as to reveal the profiles of cerebral proteins related to its effectiveness in improving cerebral ischemia (CI). METHODS: SD rats were rando-mized into sham operation (sham), CI model, non-acupoint and acupoint groups (n=10 in each group). The CI model was established by MCAO according to modified Longa's method. For rats of the acupoint group, "Dazhui" (CV 14), "Baihui" (GV 20) and "Shuigou" (GV 26) were punctured by twirling the filiform needles for about 1 min, and repeated once again during 30 min of need-le retention, which was conducted once every 12 h, and 6 times altogether. The non-acupoints were about 3 mm away from the above-mentioned acupoints and stimulated with the same method and same procedures. The neurological deficit score was scaled according to Longa's method. The differentially-expressed proteins (≥ 1.5 folds in up- and down-regulation) in the ischemic region of the brain were detected by using Springbio 720 antibody chip technology. RESULTS: Compared with the sham group, the neurological deficit score was significantly higher in the model group (P<0.01), while compared with the model group, the neurological deficit scores were considerably in both acupoint and non-acupoint groups (P<0.05, P<0.01). The therapeutic effect of acupoint group was evidently superior to that of the non-acupoint group in improving neurological function (P<0.05). Compared with the sham group, the differentially-expressed proteins in the ischemic brain including 33 up-regulated and 12 down-regulated were found in the model group, mainly functioning in cell apoptosis, cell cycle regulation, cell differentiation and proliferation, cell cytoskeleton and connection, cell signal transduction, DNA repair and transcription factors. Compared with the model group, the differentially-expressed proteins including 12 up-regulated and 31 down-regulated in the acupoint group, and 15 up-regu-lated and 17 down-regulated in the non-acupoint group were detected, functioning being the same as those mentioned above in the model group. CONCLUSIONS: Manual acupuncture stimulation of GV 14, GV 20 and GV 26 can improve neurological function in CI rats, which may be associated with its function in regulating the expression of many proteins in the ischemic region of the brain.

[Acupuncture treatment of male infertility: a systematic review].

OBJECTIVE: To systematically evaluate acupuncture as a treatment for male infertility. METHODS: We searched Chi na Biology Medical Database (CBM), Wan Fang Medical Information System, China National Knowledge Infrastructure (CNKI), VIP Information Resource System (VIP), and PubMed for published literature on acupuncture as a treatment for male infertility on May 1 2014. Based on the Standards for Reporting Interventions in Clinical Trials of Acupuncture (STRICTA), we evaluated the quality of the reports, conducted meta-analysis on the identified studies via RevMan5.2, and assessed the quality of the evidence in the literature by Grading of Recommendations Assessment, Development, and Evaluation (GRADE). RESULTS: A total of 12 studies involving 2,177 patients were included, the quality of which was evaluated as mediocre. With regard to the cure rate, acupuncture was comparable to traditional Chinese medicine (TCM) (P > 0.05) but better than Western medicine (RR = 4.00, 95% CI 1.63 to 9.82, P < 0.01) while acupuncture + TCM was better than either TCM (RR = 1.77, 95% CI 1.20 to 2.60, P < 0.01) or Western medicine used alone (RR = 2.73, 95% CI 1.51 to 4.93, P < 0.01), and acupuncture + Western medicine was better than Western medicine alone (RR = 1.88, 95% CI 1.17 to 3.02, P = 0.01). The combined use of acupuncture, ear pressure, TCM, and Western medicine showed a higher cure rate than the combination of TCM and Western medicine (RR = 3.45, 95% CI 2.90 to 4.11, P < 0.01). In therapeutic effectiveness, acupuncture was comparable to TCM (P > 0.05) but superior to Western medicine (RR = 1.41, 95% CI 1.12 to 1.71, P < 0.01), acupuncture + TCM was superior to either TCM (RR = 1.14, 95% CI 1.05 to 1.23, P < 0.01) or Western medicine alone (RR = 1.43, 95% CI 1.22 to 1.67, P < 0.01), and acupuncture + Western medicine was superior to Western medicine alone (RR = 1.25, 95% CI 1.05 to 1.49, P = 0.01). In improving sperm concentration, acupuncture was as effective as TCM (P > 0.05) and sham acupuncture (P > 0.05) but outdid Western medicine (RR = 27.00, 95% CI 24.27 to 29.73, P < 0.01) and acupuncture + TCM outdid either TCM (RR = 14.65, 95% CI 7.58 to 21.72, P < 0.01) or Western medicine alone (RR = 1.04, 95% CI--1.43 to 3.51, P > 0.05). In improving grade a sperm, acupuncture exhibited a similar effect to TCM (P > 0.05) and sham acupuncture (P > 0.05), and acupuncture + TCM was more effective than TCM alone (RR = 7.78, 95% CI 3.51 to 12.23, P < 0.01) but equally effective as Western medicine (P > 0.05). In elevating the level of grade a + b sperm, acupuncture + TCM excelled either TCM (RR = 11.00, 95%, CI 3.17 to 18.82, P < 0.01) or Western medicine alone (RR = 12.22, 95% CI 6.87 to 17.57, P < 0. 01), while acupuncture produced a comparable effect with sham acupuncture (P > 0.05). As for the quality of the included studies, only 3 conclusions of the 23 meta-analyses were assessed to be of average quality, while the others of poor or extremely poor quality. Therefore, the recommendation grade of the conclusions was low. CONCLUSION: For the treatment of male infertility, acupuncture is reported to be equally effective as TMC and more effective than Western medicine, and its effectiveness is enhanced when applied in combination with either TCM or Western medicine. Acupuncture is distinctively efficacious in improving sperm quality. Nevertheless, the overall quality of the included studies is low.

Effects of electroacupuncture at Shangjuxu (ST 37) on interleukin-1beta and interleukin-4 in the ulcerative colitis model rats.

OBJECTIVE: To probe into the mechanisms of electroacupuncture (EA) at Shangjuxu (ST 37) for treatment of the ulcerative colitis (UC). METHODS: Forty male Wistar rats were randomly divided into 4 groups: a normal group, a model group, a Shangjuxu group and a non-acupoint group, 10 rats in each group. The UC rat model was made with enema of trinitro-benzenesulfonic acid (TNBSA), and the changes of interleukin-1beta (IL-1beta) and interleukin-4 (IL-4) contents after EA at Shangjuxu (ST 37) were observed. RESULTS: EA at Shangjuxu (ST 37) could significantly decrease the IL-1beta content and increase the IL-4 content in the colic tissues of the UC rats with significant differences as compared with the model group and the non-acupoint group (P<0.05 or P<0.01). CONCLUSION: The mechanisms of EA at Shangjuxu (ST 37) for treatment of the UC rats is possibly related with the decrease of IL-1beta, a inflammation-promoting cytokine, and the increase of IL-4, a anti-inflammatory cytokine.