Effects of Electroacupuncture at Shangjuxu （ST 37） on lnterleukin-1β 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 interleuldn-1β （IL-1β） 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-1β 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-1β, a inflammation-promoting cytokine, and the increase of IL-4, a anti-inflammatory cytokine.     

Effects of Electroacupuncture on Expressions of Angiogenesis Factors and Anti-angiogenesis Factors in Brain of Experimental Cerebral Ischemic Rats after Reperfusion

Objective： To explore the mechanism of electroacupuncture （EA） in improving ischemic stroke. Methods： The Wistar rat model of focal cerebral ischemia-reperfusion was prepared by the thread embolism method. The rats were randomly divided into a normal group, a model group, and an EA group. EA was given at bilateral ＂Hegu＂ （LI 4） in rats of the EA group. Expression of the vascular endothelial growth factor （VEGF） mRNA was detected with hybridization in situ, and expressions of the angiogenin-1 （Ang-1） and endostatin proteins were detected with the immunohistochemical method. Results： As compared with the normal group, the expressions of VEGF mRNA, Ang-1 protein and endostatin protein significantly increased in the model group （all P〈0.05）; and when compared with the model group, the EA group showed even more significant increase in expressions of the VEGF mRNA and Ang-1 protein （both P〈0.05）, but with an obvious decline in the increase of expression of endostatin protein （P〈0.05）. Conclusions： EA can promote angiogenesis in brain of experimental cerebral ischemic rats after reperfusion probably through up-regulating the expression of angiogenesis factors and down-regulating the expression of anti-angiogenesis factors.     

Effect of catgut implantation at acupoints on GABA. and mGluR1 expressions in brain stem of rats with spasticity after stroke

OBJECTIVE： To investigate the effect of catgut implantation at acupoints on the expressions of γ-amino butyric acid B receptor（GABAB ） and metabotropic glutamate receptor 1（mGluR1） in the brain stem of rats with spasticity after stroke.METHODS： In total, 60 male Sprague-Dawley rats were randomly divided into three groups： a sham group（n=10）, a model group（n=25） and a treatment group（n=25）. The rats in both the model group and the treatment group were subjected to middle cerebral artery occlusion to establish a model of focal cerebral ischemia. Rats with limb-spasm met the inclusion criteria. Only the left carotid artery was isolated in sham group rats. Three days after modeling, the treatment group was subjected to catgut implantation at Dazhui（GV 14）, Guanyuan（CV 4）, and Zhongwan（CV 12）. Neurological deficit symptoms were assessed with the Zea-Longa neurological deficit score. The Modified Ashworth Scale（MAS）, and isolated muscle tone were used to evaluate spasticity before and after treatment. Immunohistochemistry was applied to determine the expression of GABAB and mGluR1 in the rat brain stem after treatment.RESULTS： After treatment, neural impairment symptoms had significantly improved in the treatment group when compared to the model group（P〈0.05）. Both MAS and isolated muscle tone in the treatment group were significantly decreased when compared with the model group（P〈0.05）,and were also lower than before treatment. GABAB expression was significantly higher and mGluR1 was lower in the treatment group when compared with the model group（P〈0.01 and P〈0.05, respectively）.CONCLUSION： Catgut implantation at Dazhui（GV 14）, Guanyuan（CV 4）, and Zhongwan（CV 12）, can relieve limb spasticity by increasing the expression of GABAB and reducing the expression of mGluR1 in the brain stem of rats after stroke.     

ELECTROACUPUNCTURE PRETREATMENT ALTERS GENOME-WIDE GENE EXPRESSIONS IN RATS WITH ACUTE ISCHEMIA-REPERFUSION HEART

Objective: Employing RNA-seq technology to study gene expression profiles in electroacupuncture-pretreated acute myocardial ischemia-reperfusion animal model.Methods: Seventy-one male SD rats were randomly divided into 4 groups: sham-operated(SO) group, model(I/R) group,electroacupuncture(EA) group and EA at non-acupoint(NA)group.The left anterior descending coronary artery(LAD) was occluded in the I/R group for 30 min followed by 4 h of reperfusion.In the SO group, the LAD was exposed but not occluded.The EA group was treated by EA at Neiguan(PC6) acupoint for2 weeks before LAD occlusion.The NA group was pretreated by EA at the end point of the trial for 2 weeks.Arrhythmia score,serum enzyme testing, and TTC staining were measured for efficacy evaluation of the treatment.Heart tissues were harvested upon sacrifice of rats and RNA-seq analysis was employed for gene expression profiling.Results: Compared with the I/R group, the survival rate in the EA group was significantly increased(78% and 59%, P0.05),whereas, the survival rate was slightly increased to 67% in the NA group.The arrhythmia score in the EA group was decreased(P0.05); LDH, CK, and CK-Mb in the EA group were significantly decreased(P0.05).Injury area was reduced significantly.RNAseq analysis showed that EA reversed expression levels of genes which were pathologically up- or down-regulated in the I/R rats.These genes are involved in 11-12 pathways.NA treatment also reversed expression levels of genes which were pathologically up- or down-regulated in the I/R rats, including 4-5 pathways.Gene pathway analysis showed that EA pretreatment reduced I/R-induced release of the apoptotic proteins cytochrome c and Endo G from mitochondria and enhanced NF-κB, Bcl2a1 d and Xiap levels.Conclusion: EA pretreatment may reverse numerous ischemiareperfusion-induced pathological gene expressions and therefore protect from myocardial injury.Anti-apoptosis might be one of the underlying potential mechanisms of EA against I/R injury.     

Regulation of Moxibustion on the Expression of NF-κBp65 and PPARγ mRNA in Colon of Rats with Ulcerative Colitis

Objective: To explore the mechanism of herb-partitioned moxibustion for ulcerative colitis(UC) through observing the colonic mucosal histopathological changes and the expression of nuclear factor kappaB(NF-kB) and peroxisome proliferators-activated receptor γ(PPARγ) mRNA of UC rats. Methods: Male SD rats were randomly divided into a normal group and a model group. UC model was established by general immunological plus local irritation method. After model identification, rats in the model group were randomly divided into a model group, a herb-partitioned moxibustion(HPM) group and a Western medicine(Salicylazosulfapyridine, SASP) group. Rats in the HPM group received treatment at bilateral Tianshu(ST 25) and Dachangshu(BL 25), two cones for each point, once a day for 7 d. SASP group rats were gavaged with SASP. The pathological scores were evaluated according to hematoxylin-eosin(HE) staining of colonic tissues. We used light microscopy to observe degree of colonic mucosal damage and the quantitative polymerase chain reaction(QPCR) to detect the expression of NF-κBp65 and PPARγ in colorectal mucosa. Results: Compared with the normal group, histopathological scores were significantly higher in the model group(P0.05); compared with the model group, the scores were decreased significantly in the HPM group and the SASP group. Compared with the normal group, NF-κBp65 mRNA expression was increased with statistical significance in the model group(P0.05); compared with the model group, NF-κBp65 mRNA expressions were decreased significantly in the HPM group and the SASP group. Compared with the normal group, PPARγ mRNA expression was increased significantly in the model group(P0.05); compared with the model group, PPARγ mRNA expressions were decreased significantly in the HPM group and the SASP group. Conclusion: HPM could improve the mucosa damage of UC rats, which is possibly through down-regulating NF-κBp65 to achieve anti-inflammatory effect. Whether decreasing the PPARγ mRNA is possibly involved in preventing precancerosis will need further study.     

Effect of Chinese Drugs for Promoting Blood Circulation and Eliminating Blood Stasls on Vascular Endothelial Growth Factor Expression in Rabbits with Glucocorticoid-induced Ischemic Necrosis of Femoral Head

Objective： To probe into the mechanism of Chinese drugs for promoting blood circulation and eliminating blood stasis in the prevention and treatment of glucocorticoid-induced ischemic necrosis of femoral head. Methods： Thirty New Zealand adult white rabbits were randomly divided into a normal control group （n=5） and a model group （n=25）. Hydroxyprednisone acetate was intramuscularly administered to the rabbits in the model group in a dosage of 7.5 mg/kg, twice per week for 6 weeks, to induce ischemic necrosis of femoral head and normal saline of the equal volume was intramuscularly administered to the rabbits in the normal control group, twice per week for 6 weeks. Then, the 5 rabbits from the normal control group and 5 rabbits selected randomly from the model group were sacrificed and the changes in histopathology and the expression of Vascular Endothelial Growth Factor （VEGF） were observed. The other 20 rabbits in the model group were randomly divided into the treatment group 1 and the treatment group 2, and the control group 1 and the control group 2, 5 rabbits in every group. Taohong Siwu Tang （桃红四物汤 Decoction of Four Drugs with Addition of Peach Kernel and Safflower） was orally administered to the rabbits in the treatment group 1 and the treatment group 2 in a dosage of 7 ml/kg, once daily and normal saline of the equal volume was orally administered to the rabbits in the control groupl and the control group, 2 once daily. After 10 weeks the rabbits in the treatment group 1 and the control groupl were sacrificed and after 13 weeks the rabbits in the treatment group 2 and the control group 2 were sacrificed, and the expression of VEGF was detected in these rabbits. Results： The expression of VEGF was significantly enhanced in rabbits of the model group as compared with the normal control group （P〈0.01）, and gradually reduced with the lapse of time. The expression of VEGF in the control groups was significantly reduced as compared with the treatment groups （P〈0.001）. Conclusion： Chinese drugs for promoting blood circulation and eliminating blood stasis can improve the microcirculation of femoral head in rabbits with glucocortieoid-induced ischemic necrosis of femoral head by promoting the expression of VEGF.     

Effect of electroacupuncture combined with stretta radiofrequency treatment on MTL and GAS in patients with gastroesophageal reflux disease

Objective： To observe the effect of electroacupuncture （EA） combined with Stretta radiofrequency treatment on contents of motilin （MTL） and gastrin （GAS） in gastroesophageal reflux disease （GERD） patients. Methods： A total of 90 eligible GERD cases were randomly allocated into three groups, 30 in each group. Patients in the EA group were treated with EA, patients in the radiofrequency group were treated with Stretta radiofrequency, and patients in the observation group were treated with EA combined with Stretta radiofrequency. Assessment was made after a course of treatment. Results： The recovery, improvement and failure cases and total effective rate in the EAgroup were 2, 18, 10 and 67.7% respectively, versus 13, 13, 4 and 86.7% in the radiofrequency group and 20, 9, 1 and 96.7% in the observation group. The therapeutic efficacy in the observation group was significantly better than that in the other two groups （P〈0.01）. After treatment, symptom scores of regurgitation, heartburn and substernal burning pain in all three groups were significantly reduced （P〈0.05）; the reduction in the observation group was more significant than that inthe other two groups （P〈0.05）,and the reduction in the radiofrequency group was more significant than that in the EA group （P〈0.05）. After treatment, the contents of MTL and GAS in all three groups were increased; and the increase in the observation group was more significant than that in the other two groups （P〈0.05）. Conclusion： EA combined with Stretta radiofrequency treatment can significantly improve the clinical effect of GERD patients, improve regurgitation, heartburn and substernal burning pain and increase the contents of MTL and GAS.     

Effects of Bushen Huoxue formula on the learning and memory function and the cerebral neurotransmitters in diabetic mice

Objective： To observe the effects of Bushen Huoxue Formula （Formula for reinforcing the kidney and activating blood circulation） on the learning and memory function and the cerebral neurotransmitters in diabetic mice. Methods： Forty ICR mice were randomized into the normal control group, model group, Nimotop group and Chinese medicine group, 10 mice in each group. Tail intravenous injection of alloxan was applied to prepare diabetic model. Four weeks later, intragastric administration of Bushen Huoxue Formula for the Chinese medicine group, Nimotop for the Nimotop group, and isometric distilled water for the other two groups were respectively given for 8 weeks. The changes in the blood sugar level were observed; the learning and memory function was detected by Morris labyrinth test; and the contents of norepinephrine （NE）, dopamine （DA）, 5-hydroxyltryptamine （5-HT） and 5-hydroxyl indole acetic acid （5-HIAA） in cerebral cortex were determined in mice of all the groups. Results： The blood sugar levels in the diabetic model mice significantly increased as compared with those of the normal control group determined 72 h and 12 weeks later （P〈0.05 or P〈0.01）. Latencies for Morris labyrinth test in the Nimotop group and the Chinese medicine group were significantly shortened as compared with that in the model group （P〈0.01）. The contents of cortical NE in the Chinese medicine group was significantly higher than that in the model group （P〈0.01）. Conclusion： Bushen Huoxue Formula can improve the learning and memory function in the diabetic mice, and the mechanism is possibly related with change of the cortical NE content.     

Therapeutic Observation of Point-towards-point Electroacupuncture for Cervical Spondylotic Radiculopathy简

Objective： To compare the therapeutic efficacies of point-towards-point electroacupuncture（EA）, EA with Jiaji points（EX-B 2）, and Jing Fu Kang in treating cervical spondylotic radiculopathy（CSR）, and to explore the optimal treatment protocol. Methods： Totally 160 patients with CSR were randomized into three groups： a point-towards-point group（n=60） treated with EA with point-towards-point method; a Jiaji group（n=60） treated with EA at cervical Jiaji（EX-B 2） points; a medicine group（n=40） treated with oral administration of Jing Fu Kang alone. The clinical efficacies were compared afterwards. Results： After treatment, the recovery rate and total effective rate of the point-towards-point group were significantly better than that of the Jiaji group and medicine group（both P〈0.01）. After 1-week treatment, the symptom and function score of the point-towards-point group was significantly better than that of the Jiaji group and medicine group（both P〈0.01）; the point-towards-point group and Jiaji group both achieved significant improvements in the symptom and function score（P〈0.01, P〈0.05）. After 2-week treatment, the three groups all achieved marked improvements in the symptom and function score（P〈0.01）. At the end of treatment, in comparing the symptom and function score, the point-towards-point group was significantly different from the medicine group（P〈0.01） and Jiaji group（P〈0.05）; the difference between the Jiaji group and medicine group was also statistically significant（P〈0.05）. Conclusion： Point-towards-point EA can rapidly improve the symptoms and function of CSR patients, and it＇s superior to EA at Jiaji（EX-B 2） and oral administration of Jing Fu Kang in comparing the clinical efficacy.     

Anti-diabetic Activity of Zhenqing Recipe and Ligustri Lucidi Fructus in Type 2 Diabetic Rats

Objective To investigate the influence of Zhenqing Recipe(ZQR)and Ligustri Lucidi Fructus(LLF)on diabetic rats and its possible mechanism.Methods The model of type 2 diabetic rats was established by feeding a high-sucrose-high-fat diet and injecting a low dose of Streptozotocin in Wistar rats.The model rats were randomly divided into three groups:diabetic model,ZQR-treated,and LLF-treated groups for 8-weeks treatment.The normal Wistar rats were as a normal control group.Results The level of fasting blood glucose in ZQR and LLF groups was decreased compared with model group(P<0.01,0.05,respectively).Both ZQR and LLF markedly reduced serum triglycerides(P<0.01,0.05,respectively),and increased the insulin sensitivity index(P<0.05).Histopathology revealed that ZQR and LLF reduced pancreatic damage.Immunohistochemistry evaluation showed that the percentage of insulin positive cells in pancreatic island was higher than model group(P<0.01,0.05,respectively).The mRNA and protein expression of SREBP-1c in pancreas were significantly decreased in ZQR and FLL group(P<0.01).Conclusion ZQR has therapeutic effect on type 2 diabetes,it ameliorates the histopathological changes of pancreas,protectsβcells,improves insulin resistance,and attenuates the expression of SREBP-1c.This study also provides the anti-diabetic evidence of FLL even its effects are weaker than ZQR.     

Effect of electroacupuncture on expressions of VEGF and CD31 in MCAO model rats

Objective

To investigate the effect of electroacupuncture (EA) at Ganshu (BL 18) and Shenshu (BL 23) on vascular endothelial growth factor (VEGF) and platelet endothelial cell adhesion molecule-1 (PECAM-1)/CD31 around the cerebral infarction focus in middle cerebral artery occlusion (MCAO) rats and the possible mechanism, thus to provide a new strategy for the treatment of cerebral ischemic stroke by acupuncture.

Methods

A total of 180 healthy male Sprague-Dawley (SD) rats were randomly divided into a sham operation group, a model group, an acupoint group and a non-acupoint group, 45 rats in each group. MCAO model was established using the modified line-embolus method in all rats except for those in the sham operation group; rats in the acupoint group were treated with EA at Ganshu (BL 18) and Shenshu (BL 23); rats in the non-acupoint group were treated with EA at the control points; rats in other 2 groups were only subjected to bundling without treatment. Ten rats in each group were randomly selected on the 3rd day, the 14th day and the 21st day after acupuncture stimulation to test the neurological function impairment. The expression levels of CD31 and VEGF were also detected.

Results

Compared with the model group and non-acupoint group, the neurological function score of the acupoint group was decreased at each time point, and the differences were statistically significant (P<0.05, P<0.01). The expressions of VEGF and CD31 in each group were the lowest on the 3rd day, reached the peak on the 14th day and still remained at high level on the 21st day. And the differences among groups were statistically significant both on the 14th day and the 21st day (P<0.05, P<0.01). Compared with the model group and the non-acupoint group, the expressions of VEGF and CD31 in the acupoint group were increased, and the differences were statistically significant (all P<0.05).

Conclusion

EA at Ganshu (BL 18) and Shenshu (BL 23) can significantly improve the neurological function score of MCAO model rats, and shows protective effect on cerebral ischemia. The protective mechanism may be related to the up-regulation of CD31 and VEGF expression around the cerebral infarction focus in the MCAO model rats and induction of angiogenesis.

     

Effect of electroacupuncture on electrogastrogram and gastric antrum ghrelin in rats with diabetic gastroparesis

Objective

To observe the effect of electroacupuncture (EA) on the electrogastrogram and gastric antrum ghrelin in rats with diabetic gastroparesis (DGP).

Methods

Fifty Sprague-Dawley (SD) rats were randomly divided into group A, group B, group C, group D and group E, 10 rats in each group. Group A was the blank control group without intervention. Group B, Group C, Group D and Group E were treated with single dose intraperitoneal injection of 2% streptozotocin (STZ), combined with 8-week high glucose and high fat diet to establish DGP rat models. Group B was the model group without treatment. Group C was the EA at acupoint group, was treated with EA at Zusanli (ST 36), Liangmen (ST 21) and Sanyinjiao (SP 6). Group D was the EA at non-acupoint group, was treated by EA at the control points of Zusanli (ST 36), Liangmen (ST 21) and Sanyinjiao (SP 6). Rats in the metoclopramide control group received 1.7% metoclopramide solution [10 mL/(kg·bw)] by gavage. Rat’s blood glucose level was measured by blood glucose meter; gastric emptying rate was detected using phenol red as a marker; the electrogastrogram was detected by BL-420F biological function system; the protein level of ghrelin was detected by enzyme-linked immunosorbent assay (ELISA); the expression of ghrelin mRNA was detected by real-time polymerase chain reaction (RT-PCR).

Results

Compared with group A, the blood glucose of group B, C, D and E were significantly increased before and after the treatment (all P<0.01); after treatment, the gastric emptying rate of group B was significantly decreased (P<0.01), the migration rates of small intestine in group B, C, D and E were significantly lower (all P<0.01), and the protein content of ghrelin in group C was significantly decreased (P<0.01); the expressions of ghrelin mRNA were significantly increased in group B, C, D and E (all P<0.01), the mean amplitudes of electrogastrogram in group B and D were significantly decreased (both P<0.01). After treatment, compared with group B, the blood glucose of group C was significantly decreased (P<0.05), the gastric emptying rate and small intestine migration rate were significantly increased in group C and E (P<0.05, P<0.01), the small intestinal migration rate was significantly increased in group D (P<0.05), the expression of ghrelin in protein and mRNA in group C was significantly lower (P<0.01), the expression of ghrelin mRNA in group E was significantly lower (P<0.05), and the mean amplitude of electrogastrogram in group C was significantly increased (P<0.05). After treatment, compared with group D, the protein and mRNA expressions of ghrelin in group C were significantly decreased (P<0.01). After treatment, compared with group E, the protein expression of ghrelin in group C was significantly decreased (P<0.01).

Conclusion

EA at Zusanli (ST 36), Liangmen (ST 21) and Sanyinjiao (SP 6) could regulate the blood glucose level of DGP model rats, enhance electrogastrogram activity, promote gastric emptying, and regulate ghrelin expression in protein and mRNA.

     

Study on the mechanisms of electroacupuncture for promoting gastrointestinal motility in rats with diabetic gastroparesis

Objective

To observe the effect of electroacupuncture (EA) at Zusanli (ST 36) on the gastrointestinal motility and the ultrastructures of pacemaker cells [the interstitial cells of Cajal (ICC)] in diabetic gastroparesis (DGP) rats and explore the mechanism of EA for DGP.

Methods

A total of 50 Sprague-Dawley (SD) rats were randomly divided into group A, group B, group C, group D and group E, with 10 rats in each group. Group A was the blank control; a single intraperitoneal injection of 2% streptozotocin (STZ) was performed in rats of group B, group C, group D and group E, with high glucose and high fat diet for 8 weeks to establish the DGP rat models. Group B was the model group and the rats did not receive any treatment; group C was EA at acupoint group and the rats received EA at Zusanli (ST 36), Liangmen (ST 21) and Sanyinjiao (SP 6); group D was EA at non-acupoint group and the rats received EA at the control points of Zusanli (ST 36), Liangmen (ST 21) and Sanyinjiao (SP 6); group E was metoclopramide group and the rats were treated by intragastric administration of metoclopramide. Blood glucose was detected using ONE TOUCH blood glucose meter; gastric emptying rate and small intestine migration rate were measured using intragastric phenol red; ultrastructures of gastric antrum ICC were detected by transmission electron microscopy.

Results

The differences of blood glucose between group B, group C, group D, group E versus that of group A were statistically significant after modeling (P<0.01); after treatment, the differences of blood glucose between group D, group E versus that of group C were statistically significant (P<0.05, P<0.01); the gastric emptying rate of rats in group B was statistically significant different from that in group A (P<0.01); the gastric emptying rate of rats in group C was statistically significant different from that in group B (P<0.01). The migration rates of rats’ small intestines in group B, group C, group D and group E were all statistically significant different from that in group A (P<0.01); the migration rate of rats’ small intestines in group C was statistically significant different from that in group B (P<0.01). The ultrastructure of rat’s ICC in group B showed apoptosis compared with that in group A; rat‘s ICC in group C had complete basement membrane, more cytoplasm mitochondria, Golgi and rough endoplasmic reticulum, showing clear structure, occasional mitochondria swelling and gap junctions with adjacent smooth muscle cells; there were no significant differences between group D, group E versus group B.

Conclusion

EA at Zusanli (ST 36) plus other acupoints can regulate the blood glucose and promote gastrointestinal motility in DGP rats, and the mechanism may be related to repairing the damaged ICC structure.

     

Scalp acupuncture improves disordered behavior in the valproic acid-induced rat model of autism: 头针改善丙戊酸钠诱导的孤独症大鼠行为学及其机制研究

ObjectivesTo investigate the underlying mechanisms of scalp acupuncture treatment (SAT) on autism spectrum disorder (ASD).MethodsThirty male Wistar rat pups that had been prenatally exposed to valproic acid sodium (VPA) were randomly divided into the VPA, VPA+acupoint, and VPA+ non-acupoint groups using the random number table method, with 10 rats in each group. Ten pups who had been prenatally exposed to saline were assigned to the control group (CG). There was no intervention in either the control or VPA groups. In the acupoint group, “Shenting (GV24),” bilateral “Benshen (GB13)” were manipulated. In the non-acupoint group, the area below the costal space was stimulated. Acupuncture stimulation lasted for 40 min, with manual twisting of the needles every 10 min, 5 days/week, with 2 days of rest per week, for a total duration of 4 weeks. After the corresponding treatments, behavioral tests (including the open field, social interaction, and Morris water maze tests) were performed to evaluate the therapeutic effects. RT-PCR and Western blotting were performed to detect the expression of neuregulin 1 (NRG1)/ErbB4.ResultsIn the open field test, the activity time spent in the central area in the VPA+acupoint group was significantly longer than that in the VPA group and VPA+ non-acupoint group (both P<0.05). The total length in the VPA+acupoint group was significantly longer than that in the VPA group (P<0.05). The number of bouts in the central area of the VPA+acupoint group was significantly higher than that of the VPA group (P<0.05). In session I of social interaction test, all experimental rats spent more time interacting with stranger 1 (all P<0.05). In session II, the CG and VPA+acupoint groups rats showed more interest in searching for new strangers, but the VPA+non-acupoint group spent more time interacting with stranger 1 than with stranger 2(all P<0.05). In the Morris water maze test, compared with the VPA group, the latency of the VPA+acupoint group was shorter (day 2, 3, 4, 5, P<0.05); compared with VPA+acupoint group, the latency of the VPA+non-acupoint group was longer (day 2, 4, P<0.05). The mean distance in the VPA+acupoint group was shorter than that in the VPA group (day 3, 5, P<0.05). The platform quadrant time of the VPA+non-acupoint group was significantly shorter than that of the VPA+acupoint group (P<0.05) (day 6). The VPA+acupoint group had more platform crossings than the VPA group (P<0.05), and the VPA+ non-acupoint group had fewer platform crossings than the VPA+acupoint group (P<0.05) (day 6). After SAT, the expression levels of NRG1 and ErbB4 proteins in the VPA+acupoint group were significantly increased than those in the VPA group (both P<0.05) in the medial prefrontal cortex (mPFC). The expression levels of NRG1 and ErbB4 proteins in VPA+non-acupoint group were significantly less than those in the VPA+acupoint group (both P<0.05) in the mPFC. After SAT, the expression levels of NRG1 and ErbB4 proteins in the VPA+acupoint group were significantly higher than those in the VPA group (both P<0.05) in the hippocampus. The expression levels of NRG1 and ErbB4 proteins in the VPA+non-acupoint group were significantly lower than those in the VPA+acupoint group (both P<0.05) in the hippocampus. After SAT, the expression levels of mRNA levels of NRG1-ErbB4 in the mPFC and hippocampus in VPA+acupoint group were significantly higher than those in the VPA group (all P<0.05). The mRNA expression levels of NRG1-ErbB4 in the mPFC and hippocampus in VPA+ non-acupoint group were significantly lower than those in the VPA+acupoint group (all P<0.05).ConclusionsOur results suggest that SAT can improve ASD-like behaviors in young rats in a VPA model of ASD. This may be related to its function in upregulating the expression of protein and gene of NRG1 and its receptor ErbB4 in the mPFC.     

荧光双重标记Cx43在大鼠“足三里”表达的实验研究

目的:研究缝隙连接蛋白Cx43在大鼠“足三里”穴的表达及其与穴位、经络的可能关系。方法:20只健康成年SD大鼠随机分为非针刺组、针刺组,每组10只。应用荧光双重标记免疫组织化学方法和激光扫描共聚焦显微镜技术对成年大鼠皮肤肌肉组织的Cx43进行定位、定量分析。结果:皮肤肌肉组织中Cx43主要在表皮上皮细胞、皮下筋膜的肥大细胞中表达;非针刺组中穴位处Cx43的表达显著高于非经非穴处,两者相比有极显著性差异(P<0.01);穴位针刺后Cx43表达显著增加,与非针刺组中穴位比较有极显著性差异(P<0.01)。结论:电针可使“足三里”穴区皮肤及肌肉组织中的Cx43表达明显增加,缝隙连接蛋白及缝隙连接可能在经络活动中起重要作用。     

Influence of electroacupuncture on COX activity of hippocampal mitochondria in senescence-accelerated mouse prone 8 mice

Objective： To observe the effect of electroacupuncture （EA） on cytochrome c oxidase （COX）activity of hippocampal mitochondria in senescence-accelerated mouse prone 8 （SAMP8） mice, and to explore the EA mechanism on Alzheimer disease （AD） in improving energy metabolic disorder.
Methods： Twelve SAMP8 mice were randomly divided into a model group and an EA group, with six in each group. Six senescence-accelerated mouse resistance 1 （SAMR1） mice were prepared as blank group. Mice in the EA group received EA on Baihui （GV 20） and Yongquan （KI 1）, once a day for 7 d as a course, altogether 3 courses with one day intervalbetween two courses. Mice in the model group and the blank group were manipulated and fixed as those in the EA group. After interventions, Morris water maze was employed to test spatial learning and memory ability to evaluate EA effect; spectrophotometry was used to detect the activity of hippocampal mitochondria COX.
Results： Compared with the blank group, mean escape latenciesof the EA group and model group were prolonged significantly in Morris water maze tests （P〈0.01）, the residue duration in the former platform quadrant significantly decreased （P〈0.01）. Compared with the model group, mean escape latencies on 1 d, 2 d and 3 d of the EA group were significantly reduced （P〈0.05）, and those on 4 d and 5 d continued the decreasing tendency （P〈0.01）, the residue duration on the former platform quadrant was significantly prolonged （P〈0.05）. The COX activity tests showed that, compared with the blank group, COX activities of the model group and the EA group were significantly decreased （P〈0.01）; compared with the model group, COX activity of the EA group was significantly elevated （P〈0.01）.
Conclusion： It＇s plausible that EA improves AD learning and memory ability by increasing mitochondria COX activity, protecting the structure and function, and improving energy metabolism.     

Effect of electroacupuncture at Ganshu (BL 18) and Shenshu (BL 23) on the expression of EphB2 protein in cortex around cerebral infracted area of rat

Objective

To observe the effect of electroacupuncture (EA) on the expression of erythropoie-tin-producing hepatocyte receptor B2 (EphB2) in the cortex around the infracted area of middle cerebral artery occlusion (MCAO) rats at different timing, and to reveal the possible mechanism of acupuncture in the treatment of cerebral ischemia.

Methods

A total of 180 male Sprague-Dawley (SD) rats were randomly divided into a sham operation group, a model group, an acupoint group and a non-acupoint group, with 45 rats in each group. Rats in each group were further divided into three subgroups: postoperative 3 d, postoperative 14 d and postoperative 21 d groups, with 15 rats in each subgroup. The MCAO model was made by the modified occlusion method. The neurological function score, 2,3,5-triphenyl tetrazolium chloride (TTC) staining, immunohistochemistry assay, immunofluorescence double labeling method and Western blot were used to detect the corresponding indicators.

Results

The neurological impairment of rats was most obvious at postoperative 3 d, and then gradually improved with time, which was more significant in the acupoint group (P<0.05). The change of infarcted volume was consistent with the neurological function impairment. The number of EphB2 positive cells (EphB2⁺) around the infarcted area was decreased significantly at postoperative 3 d, and then gradually improved with time, which returned to the same level as that in the sham operation group at postoperative 21 d. The increase was most significant in the acupoint group (P<0.05), and the positive cell number was higher than that in the sham operation group (P<0.01). Western blot and immunohistochemistry results were basically consistent. Immunofluorescence displayed that EphB2⁺ and postsynaptic density-95 positive (PSD-95+) were co-expressed, after the MCAO operation, in the cortical neuron around the infracted area, and the number of co-expressing cells was increased gradually with time, which was most significant in the acupoint group (P<0.05).

Conclusion

Electroacupuncture at Ganshu (BL 18) and Shenshu (BL 23) can significantly improve the neurological function and cerebral infarcted volume ratio of MCAO rats, which may be related to the activation of EphB2 expression in cortex around the infracted area and the promotion of synaptic remodeling.

     

Clinical observation on electroacupuncture for post-stroke flaccid paralysis

Objective:To observe the clinical efficacy of electroacupuncture (EA) in treating post-stroke flaccid paralysis. Methods:Forty patients with post-stroke flaccid paralysis were randomized by the random number table into a treatment group and a control group, 20 cases in each. The treatment group was intervened by acupuncture at Jiquan (HT 1), Tianquan (PC 2), Ququan (LR 8), Yinlingquan (SP 9), and Yongquan (KI 1), and the control group was treated by acupuncture with conventional acupoint selection. Barthel index (BI) was adopted for evaluating the activities of daily living (ADL), and therapeutic efficacy was analyzed. Results:The two groups both had marked increases of BI score after treatment. Compared to the control group after 1 treatment course and 2 treatment courses respectively, the treatment group had significant differences in BI score (P<0.01). The total effective rate was 100.0% in the treatment group versus 90.0% in the control group, and the difference was statistically significant (P<0.01). Conclusion:EA at Jiquan (HT 1), Tianquan (PC 2), Ququan (LR 8), Yinlingquan (SP 9), Yongquan (KI 1) is an effective approach in treating post-stroke flaccid paralysis.     

Diverse needling methods for dry eye syndrome: A randomized controlled study

Objective:To explore a proper acupuncture treatment protocol for dry eye syndrome(DES),by comparing the therapeutic effects between ordinary acupuncture and electroacupuncture(EA).Methods:Forty-seven eligible subjects with DES were randomized into an acupuncture group(n=23)and an EA group(n=24).With the same acupoint formula,the acupuncture group was treated with ordinary acupuncture,and the EA group was treated with ordinary acupuncture plus electrical stimulation.After a treatment course,eye symptom score,Schirmer I test(SIT),Break-up Time(BUT)of tear film,Corneal Fluorescein Staining(CFS),and Visual Analogue Scale(VAS)were adopted in evaluation and comparison of the two groups.Results:The total effective rate was 79.2%in the EA group versus 56.5%in the acupuncture group,and the difference was statistically significant(P<0.05).After treatment,both groups had marked improvements in eye symptom score,SIT,BUT,CFS,and VAS values(P<0.001);the EA group was better than acupuncture group in improving eye symptom score and SIT value(both P<0.05);the differences were insignificant in comparing VAS,BUT and CFS results between the two groups(P>0.05).Conclusion:Both EA and ordinary acupuncture are effective in treating DES,but EA is better than ordinary acupuncture in improving eye symptom and SIT score.