Moxibustion at ST36 activates peritoneal macrophages in CTX-induced immunosuppression in mouse model via IFN-γ

Background:To explored whether moxa cone moxibustion can reduce peritoneal inflammation by increasing the content of peritoneal macrophages and B cells via interferon-gamma.Methods:The mice were randomly divided into three groups with six mice in each group:the control group,model group,and moxibustion group,and the model was established in mice using cyclophosphamide.In the moxibustion group,the mice received moxa cone moxibustion at Zusanli(ST36)for 7 days.Analysis of Peritoneal cell were detected by flow cytometry and immunofluorescence,the protein expression level in the peritoneal fluid were measured with mouse cytokine antibody arrays and verified by enzyme linked immuno sorbent assay test,and RNA-Sequencing was used for peritoneal cell RNA analysis.Results:Our results showed that moxa cone moxibustion could reduce the loss of large peritoneal macrophages and B1 cells(P<0.05).With the cytokine array analysis and enzyme linked immuno sorbent assay test of peritoneal fluid,we found that IFN‐γwas up-regulated in moxibustion group(P<0.05).There were 169 genes were down-regulated in the model group and up-regulated in the moxibustion group while 19 genes that were up-regulated in the model group and down-regulated in the moxibustion group via RNA-sequencing.Kyoto Encyclopedia of Genes and Genomes pathway analysis of 188 intersect differentially expressed genes were found that the top 3 pathways with the highest enrichment of up-regulated genes included Hematopoietic cell lineage,Inflammatory bowel disease and Malaria.The differentially expressed genes visualization protein-protein interaction network shows the top 10 genes including Ifng,Grb2,CCR7,CTLA4,CXCR5,Foxp3,kit,PRF1,CD5 and klrg1.Conclusion:These findings showed that moxa cone moxibustion can alleviate chemotherapy-induced diarrhea by reducing the loss of large peritoneal macrophages and B1 cells in the peritoneal cavity,possibly through up-regulating inflammatory bowel disease signaling pathway via interferon-gamma to regulate the survival and function of large peritoneal macrophages and B1 cells.

Comparison of protective effects of electroacupuncture and moxibustion at Zusanli(ST 36)on perinatal nicotine exposure-induced lung phenotype in rat offspring

Objective:To compare the effects of electroacupuncture(EA)and moxibustion at Zusanli(ST 36)on the lung phenotype of rat offspring exposed to nicotine during the perinatal period.Methods:Pregnant Sprague-Dawley rats were randomly divided into 4 groups:the control group(saline only),the model group(nicotine only),the EA group(nicotine+EA at ST 36 acupoints bilaterally),and the moxibustion group(nicotine+moxibustion at ST 36 acupoints bilaterally).n=6 rats per group.On postnatal day 21,the body weight,lung weight,and pulmonary function were determined and lung morphometry was performed.Peroxisome proliferator-activated receptor gamma andβ-catenin levels in the lung tissue of offspring were also determined.Results:Perinatal nicotine exposure(PNE)results in decreased body and lung weights of offspring rats,abnormal lung tissue morphology,and significantly altered pulmonary function,showing an increase in total airway resistance and a decrease in tidal volume,minute ventilation,total airway compliance,and peak expiratory flow.Bilateral EA at ST 36 acupoints could block all of these perinatal nicotine-induced effects.Although moxibustion also had protective effects in nicotine-induced offspring lungs,some of these effects did not reach statistical significance,e.g.,protection against the upregulation ofβ-catenin,the downregulation of PPARγsignaling,and the increase in peak expiratory flow.Conclusion:Maternal EA at ST 36 blocked the PNE-induced changes in key developmental signaling pathways,prevented the PNE-induced changes in lung morphology,and protected pulmonary function.Moxibustion at ST 36 showed similar but weaker protective effects against the PNE-induced changes in the exposed offspring.It is important to note that the mechanism underlying the protective effects of moxibustion at ST 36 may be different from those of EA at ST 36,and further research is needed to understand these differences.

Effect of electroacupuncture at different acupoints in treating rapid gastric emptying:Zusanli(ST36)plus Zhongwan(CV12)versus Zhongwan(CV12)alone

OBJECTIVE:To evaluate the effect of electroacupuncture on rapid gastric emptying by comparing the effectiveness of electroacupuncture at both Zusanli(ST36)and Zhongwan(CV12)(He-Mu acupoints)versus electroacupuncture at Zhongwan(CV12)alone.METHODS:Sixty-five patients were randomly divided into two groups;33 patients in the He-Mu acupoints group received electroacupuncture at both Zusanli(ST36)and Zhongwan(CV12),while 32 patients in the single acupoint group received electroacupuncture only at Zhongwan(CV12).Both groups were treated once daily for 30 min,five times per week for 3 weeks.Before and after the 3-week treatment course,patients underwent gastric emptying scintigraphy and were assessed using the Traditional Chinese Medicine(TCM)symptom scale of gastrointestinal diseases.The two groups were compared regarding the percentages of gastric retention at 30 and 60 min and the TCM symptom scores.RESULTS:After the treatment course,the percentages of gastric retention at 30 and 60 min were significantly improved in both groups compared with the pre-treatment values(P<0.01).The improvement in the gastric retention was significantly better in the He-Mu acupoints group than the single acupoint group at 30 min(P<0.01)and 60 min(P<0.05).The TCM symptom score was significantly decreased after the treatment course in both groups(P<0.01).The improvement in the TCM symptom score was significantly better in the He-Mu acupoints group than the single acupoint group(P<0.01).The total effective rate was 93.55%(29/31)in the He-Mu acupoints group and 77.42%(24/31)in the single acupoint group.The treatment effect was better in the He-Mu acupoints group than the single acupoint group(P<0.05).CONCLUSIONS:Electroacupuncture at both Zusanli(ST36)and Zhongwan(CV12)or at Zhongwan(CV12)alone is effective in treating rapid gastric emptying.Furthermore,electroacupuncture at both Zusanli(ST36)and Zhongwan(CV12)is more effective than electroacupuncture at Zhongwan(CV12)alone.

Effect of scar-producing moxibustion at the acupoints Zusanli(ST 36) and Feishu(BL 13) on neutrophil-to-lymphocyte ratio and quality of life in patients with non-small-cell lung cancer: A randomized,controlled trial

OBJECTIVE: To evaluate the effect of heat stimulation via scar-producing moxibustion at the acupoints Zusanli(ST 36) and Feishu(BL 13) on the neutrophil-to-lymphocyte ratio(NLR) and quality of life in patients with non-small-cell lung cancer(NSCLC).METHODS: Seventy patients with NSCLC were randomly assigned into two groups: group A received scar-producing moxibustion at the acupoints Zusanli(ST 36) and Feishu(BL 13) every day for 6 weeks, while group B received no intervention(control group). Outcome measures were the NLR and the scores from the European Organization for Research and Treatment of Cancer Quality of Life Questionnaire(EORTC QLQ-C30). The NLR and the EORTC QLQ-C30 were assessed at baseline and at the end of 6 weeks.RESULTS: Five participants dropped out, leaving afinal total of 65 participants who completed the trial. Groups A and B had a similar mean NLR at baseline. After the treatment course, the NLR in group A was significantly lower than that in group B(P <0.001). Compared with group B, the EORTC QLQ-C30 scores in group A were significantly greater in terms of global health status or quality of life(P < 0.001) and function(P < 0.05), and significantly lower in terms of symptoms(P < 0.05).CONCLUSION: The present study suggests that performing scar-producing moxibustion by heat-stimulating the acupoints Zusanli(ST 36) and Feishu(BL13) effectively decreases the NLR and improves the quality of life in patients with NSCLC.

Effect of dexamethasone injection into Zusanli(ST 36) acupoint on ovalbumin-induced allergic rhinitis

OBJECTIVE:To investigate the effects of acupunc ture with dexamethasone(A.Dex)on allergic rhini tis(AR)by injecting dexamethasone into the Zusan li(ST 36)acupoint.METHODS:Thirty 6-week-old female BALB/c mice were sensitized on days 1,5,and 14 by intraperitoneal injection of 100 pg of ovalbumin(OVA).The mice were then randomly divided into six groups(n =5 in each group).Five groups were sensitized intranasally with 2μL of 1.5 mg of OVA for 10 consecutive days,while one group was sensitized intranasally with PBS in a similar manner as a negative control group.One hour before each administration of intranasal OVA,two groups were orally administered either a control vehicle(distilled water;AR control group)or 200 pg/kg Dex(O.Dex group),while three groups received A.Dex at Zusanli(ST36)with Dex concentrations of 2,20,and 200μg/kg for each group,respectively.AR symptoms were evaluated by measuring the rubbing score,which comprised the number of nose,ear,and eye rubs that occurred in the initial 10 min after OVA intranasal provocation on the 10 th day.We isolated spleen,serum,and nasal mucosal tissue after measuring the rubbing score.Spleen weight was measured using an electronic microbalance.The levels of IgE,thymic stromal lymphopoietin,tumor necrosis factor-a,intercellular adhesion molecule-1,and macrophage-inflammatory protein-2 in serum or nasal mucosal tissue were measured using enzyme-linked immunosorbent assays.The serum histamine levels of OVA-sensitized AR mice were measured using O-phthaldialdehyde spectrofluorometry.Western blot analysis was performed on nasal mucosal tissue extracts.RESULTS:A.Dex significantly reduced the rubbing score,spleen weight,serum IgE,and serum histamine in OVA-sensitized mice.A.Dex significantly decreased the serum levels of inflammatory cytokines(thymic stromal lymphopoietin and tumor necrosis factor-a)in OVA-sensitized mice.A.Dex significantly reduced the nasal mucosal levels of inflammatory markers(intercellular adhesion molecule-land macrophage-inflammatory protein-2)in AR mice.A.Dex effectively attenuated the expression of caspase-1 and receptorinteractingprotein-2 in nasal mucosal tissue.CONCLUSION:A.Dex may be a new and useful therapy for AR.

TREATMENT WITH ACUPUNCTURE AT ZUSANLI (St 36) FOR EPIGASTRIC PAIN IN THE ELDERLY

Acupuncture at a single point is a spe-cial feature in the treatment of acute symp-toms for its simplicity and effectiveness.Theauthor experienced many cases of acuteepigastric pain treated with acupuncture at asingle acupoint with satisfactory results.Pain in the elderly is most often of

Effects of electroacupuncture at Zusanli(ST36) on inflammatory cytokines in a rat model of smokeinduced chronic obstructive pulmonary disease

OBJECTIVE： Improvement in lung function was reported after acupuncture treatment of chronic obstructive pulmonary disease （COPD）, but little is known about the underlying mechanisms. Because an immune response imbalance could be seen in COPD, we hypothesize that electroacupuncture （EA） may play a role in regulating inflammatory cytokines and contribute to lung protection in a rat model of smoke-induced COPD. METHODS： A COPD model using male Sprague-Dawley rats exposed to cigarette smoke was established. The rats were randomly divided into four groups （control, sham, COPD, and COPD plus EA）, and COPD model was evaluated by measuring pulmonary pathological changes and lung function. EA was applied to the acupuncture point Zusanli （ST36） for 30 min/d for 14 d in sham and COPD rats. Bronchoalveolar lavage fluid （BALF） was used to measure levels of tumor necrosis factor-a （TNF-a）, interleukin-113 （IL-113）, and malonaldehyde （MDA）. RESULTS： Compared with the control rats, COPD rats had significant changes in lung resistance （R,） and lung compliance （C,） （both P〈0.01）, bronchi and bronchiole airway obstruction （P〈0.01）, and levels of MDA, TNF-α, and IL-1β（P〈0.01）. There were no significant differences between the control and the sham groups. Compared with the COPD rats, the COPD plus EA rats had decreased R, and increased CL （both P〈0.05）, and reduced bronchi and bronchiole airway obstruction （P〈0.05, P〈0.01, respectively）, while levels of TNF-α, IL-1β, and MDA in BALF were lowered （P〈0.05 and P〈0.01, respectively）. However, TNF-α and IL-1β levels of the EA group rats remained higher than those of the control group （P〈0.05）. CONCLUSION： EA at ST36 can reduce lung injury in a COPD rat model, and beneficial effects may be related to down-regulation of inflammatory cytokines. The anti-inflammatory and antioxidant effects may prolong the clinical benefit of EA.

Electro-acupuncture at Zusanli(ST 36) to improve lower extremity motor function in sensory disturbance patients with cerebral stroke A randomized controlled study of 240 cases

BACKGROUND： Studies have shown that sensory transduction is a way to introduce needle sensation. OBJECTIVE： To observe the influence of electro-acupuncture at the ＂Zusanll＂ （ST 36） point on lower extremity motor function in various sensory disturbance patients with cerebral stroke. DESIGN, TIME AND SETTING： A randomized, controlled, clinical study was performed at the Department of Neurological Rehabilitation, China Rehabilitation Research Centre from September 2006 to June 2008. PARTICIPANTS： Patients with first-time cerebral infarction or hemorrhage, or with a stroke history, but no neurodysfunction （single damage）, were selected for this study. The subjects were right-handed and disease state was stable. A total of 240 inpatients were randomly assigned to the following groups： electro-acupuncture （n = 124） and control （n = 116）. The two groups were further assigned into sub-groups： no sensory disturbance, superficial sensory disturbance, deep sensory disturbance, and deep and superficial sensory disturbance. METHODS： On the basis of routine limb function training, the acupoint Zusanliwas utilized in all patients from the electro-acupuncture group. Perpendicular acupuncture was 3.0-4.0 cm deep. An electric acupuncture instrument was connected when patients developed the needle sensation, deqi. A stimulation pattern consisting of distant and dense waves of 50 Hz was used to elicit slight dorsal foot extension. Acupuncture was administered 5 times per week, 30 minutes per session, for 6 weeks in total. MAIN OUTCOME MEASURES： FugI-Meyer assessment （FMA） was used to evaluate lower extremity motor function; Ver.1.0 gait analysis to estimate gait （step frequency, step speed, and step scope）; lower extremity Composite Spasticity Scale （CSS） to estimate muscle spastic degree. RESULTS： Following treatment, motor function improved in both groups. Compared with the control group, FMA score, step speed, step frequency, and step scope were increased in the electro-acupuncture group, but there was no difference in lower extremity CSS scores between the electro-acupuncture and the control groups （P 〉 0.05）. Compared with the control group, Zusanli （ST 36） electro-acupuncture improved motor function indices as follows： FMA score, step frequency, step speed, and step scope of patients with no sensory disturbance （P 〈 0.05-0.01）, step frequency of patients with superficial sensory disturbance （P 〈 0.05）, and step frequency and step speed of patients with deep sensory disturbance （P 〈 0.05）. CONCLUSION： Zusanfi （ST 36） electro-acupuncture effects on lower extremity motor function in stroke patients were improved with no muscle tone rise. Therefore, this form of treatment can be used in convalescent treatment, Moreover, effects were different according to various sensory disturbance types, which suggested that sensory input influenced acupuncture effects.

Effects of pre-moxibustion at Zusanli (ST36) on heat shock protein 70 expression in rats with gastric mucosal lesions after neurotomy

Studies have shown that pre-moxibustion protects the gastric mucosa by up-regulating the expression of heat shock protein 70. However, the signaling pathway underlying this effect remains unclear. Rats were intragastrically administered absolute alcohol, causing obvious lesion of the gastric mucosa. Following pre-moxibustion at Zusanfi （ST36） for 8 days, the ulcer index decreased to different degrees. The results of an enzyme linked immunosorbent assay and western blotting showed significant upregulation of heat shock protein 70 expression in the gastric mucosa and serum. None out of transection of the spinal cord, damage to the nucleus of the solitary tract, neurotomy of the vagal nerve and neurotomy of the common peroneal nerve affected the decrease in ulcer index or the increase in heat shock protein 70 expression in serum after pre-moxibustion at Zusanfi, and heat shock protein 70 expression was obviously decreased in the gastric mucosa. These findings suggest that pre-moxibustion at Zusanfi can protect the gastric mucosa against lesioning, and that the mechanism underlying this effect involves its induction of heat shock protein 70 expression. Neural pathways participate in the regulatory effects of moxibustion on heat shock protein 70 expression in the gastric mucosa.

EXPERIMENTAL STUDY ON MOXIBUSTION AT ZUSANLI(ST 36) AND LIANGMEN (ST 21) INDUCING HEAT SHOCK PROTEIN 70 (HSP70) EXPRESSION TO RESIST OXIDATIVE INJURY OF GASTRIC MUCOSA

Objective: To observe effect of moxibustion at Zusanli (足三里 ST 36) and Liangmen (梁门 ST 21) on expression of heat shock protein 70 (HSP70) in gastric mucosa of the rat of stress ulcer (SU) to explore the mechanism of moxibustion in resisting oxidative injury of the gastric mucosa. Methods: Sixty SD rats were evenly randomized into 4 groups, a blank group, a model group, an acupoint moxibustion group and a non-acupoint moxibustion group. Water restraint stress (WRS) method was used to make stress gastric ulcer rat model. The ulcerative index (UI) of gastric mucosa was evaluated by using GUTH method, the gastric mucosa blood flux (GMBF) was detected by a laser Doppler bloodflow monitor, and HSP70 expression and malondialdehyde (MDA) content in the gastric mucosa were determined respectively with immunohistochemical and thiobarbiturate methods. Results: Moxibustion at Zusanli (ST 36) and Liangmen (ST 21) significantly decreased UI, up-regulated HSP70 expression, increased GMBF, and decreased MDA content in the gastric mucosa in the rat of stress gastric ulcer, with significant differences as compared with the model group and the non-acupoint moxibustion group (P<0.05 or P<0.01). Conclusion: Moxibustion at Zusanli (ST 36) and Liangmen (ST 21) can induce high expression of HSP70 and decrease MDA content in the gastric mucosa, so as to resist oxidative injury, with relative acupoint specificity.

Effect of Sanhuangwuji powder,anti-rheumatic drugs,and ginger-partitioned acupoint stimulation on the treatment of rheumatoid arthritis with peptic ulcer:a randomized controlled study

OBjECTIVE:To observe the efficacy and safety of oral Sanhuangwuji powder,anti-rheumatic drugs(ARDs),and ginger-partitioned acupoint stimulation at Zusanli(ST 36) on the treatment of rheumatoid arthritis(RA) complicated by peptic ulcer.METHODS:This prospective randomized controlled study included 180 eligible inpatients and outpatients randomly assigned to an ARD treatment(n = 60),ginger-partitioned stimulation(n =60),or combination treatment(n = 60).Patients assigned to the ARD group were given oral celecoxib,methotrexate,and esomeprazole.Patients assigned to theginger-partitioned stimulation group were given ginger-partitioned acupoint stimulation at Zusanli(ST 36) in addition to the ARDs.Patients in the combination treatment group were given oral Sanhuangwuji powder,ginger-partitioned acupoint stimulation at Zusanli(ST 36),and ARDs.All patients were followed up for 2 months to evaluate clinical effects and safety.The study was registered in the World Health Organization database at the General Hospital of Chengdu Military Area Command Chinese People's Liberation Army(ChiCTR-TCC12002824).RESULTS:The combination treatment group had significantly greater improvements in RA symptoms,laboratory outcomes,and gastrointestinal symptom scores,compared with the other groups(P < 0.05).The peptic ulcer healing rate in the combination treatment group was significantly greater than that in the ARD treatment group(x^2 = 16.875,P < 0.05) and the ginger-partitioned stimulation group(x^2 = 6.171,P<0.05).CONCLUSION:Combination treatment with ginger-partitioned acupoint stimulation at Zusanli(ST36),oral Sanhuangwuji powder,and ARDs had a better clinical effect for RA with complicated peptic ulcer,compared with ARD treatmentalone or in combination with ginger-partitioned acupoint stimulation.

中药内服联合电针对行Halsted疝修补术的老年腹股沟嵌顿疝患者胃肠功能的影响

目的:观察中药内服联合电针对行Halsted疝修补术的老年腹股沟嵌顿疝患者胃肠功能的影响。方法:将64例行Halsted疝修补术的老年腹股沟嵌顿疝患者随机分为治疗组和对照组,每组32例。对照组给予常规治疗,治疗组在对照组治疗的基础上给予中药内服联合电针足三里治疗。观察两组患者肠鸣音恢复时间、肛门排气时间、排便时间、胃肠减压时间及术后围手术期腹胀发生率等。结果:治疗组肠鸣音恢复时间、肛门排气时间、排便时间、胃肠减压时间短于对照组,差异有统计学意义(P<0.05);治疗组腹胀发生率为18.75%,对照组腹胀发生率为53.13%,治疗组腹胀发生率低于对照组,差异有统计学意义(P<0.05)。结论:中药内服联合电针能够促进行Halsted疝修补术的老年腹股沟嵌顿疝患者胃肠功能及早恢复,降低腹胀发生率,减轻腹胀程度。

Long-term acupuncture treatment has a multitargeting regulation on multiple brain regions in rats with Alzheimer＇s disease：a positron emission tomography study

The acute effect of acupuncture on Alzheimer＇s disease,i.e.,on brain activation during treatment,has been reported.However,the effect of long-term acupuncture on brain activation in Alzheimer＇s disease is unclear.Therefore,in this study,we performed long-term needling at Zusanli（ST36）or a sham point（1.5 mm lateral to ST36）in a rat Alzheimer＇s disease model,for 30 minutes,once per day,for 30 days.The rats underwent 18F-fluorodeoxyglucose positron emission tomography scanning.Positron emission tomography images were processed with SPM2.The brain areas activated after needling at ST36 included the left hippocampus,the left orbital cortex,the left infralimbic cortex,the left olfactory cortex,the left cerebellum and the left pons.In the sham-point group,the activated regions were similar to those in the ST36 group.However,the ST36 group showed greater activation in the cerebellum and pons than the sham-point group.These findings suggest that long-term acupuncture treatment has targeted regulatory effects on multiple brain regions in rats with Alzheimer＇s disease.

经皮穴位电刺激治疗下肢动脉硬化闭塞患者的临床观察

目的探讨经皮穴位电刺激(Transcutaneous electrical acupoint stimulation,TEAS)治疗下肢动脉硬化闭塞症(Arteriosclerosis obliterans,ASO)的临床疗效。方法选取2021年3月—2022年10月期间南昌大学第一附属医院收治的98例ASO患者,按随机数字表法分为TEAS组和对照组,每组各49例。对照组予以常规治疗:包括控制饮食,低盐低脂饮食、控制血压、降血脂治疗,TEAS组给予TEAS治疗。治疗3周后,观察比较两组患者的临床疗效、不良反应情况,治疗前后的临床症状积分、踝肱指数(Ankle brachial index,ABI)和视觉模拟评分(Visual analogue scale score,VAS)、血清白细胞介素-6((Interleukin-6,IL-6)、肿瘤坏死因子-α(Tumor necrosis factor-α,TNF-α)、β2微球蛋白(β2 microglobulin,β2-M)水平。结果治疗后两组患者各临床症状积分及总积分均明显低于治疗前,差异有统计学意义(P<0.05);且TEAS组各临床症状积分及总积分较对照组明显降低,差异有统计学意义(P<0.05)。治疗后两组患者ABI值均明显高于治疗前,VAS评分明显低于治疗前,差异有统计学意义(P<0.05);且TEAS组ABI值较对照组明显升高,VAS评分较对照组明显降低,差异有统计学意义(P<0.05)。治疗后两组患者血清TNF-α、IL-6、β2-M水平均明显低于治疗前,差异有统计学意义(P<0.05);且TEAS组血清TNF-α、IL-6、β2-M水平较对照组明显降低,差异有统计学意义(P<0.05)。TEAS组的治疗总有效率(93.88%)高于对照组(73.47%),差异有统计学意义(P<0.05)。治疗期间,两组患者均未发生严重不良事件。结论TEAS治疗ASO能够有效改善下肢缺血,抑制炎症反应,减轻肢端麻木、酸胀、发冷等症状,减轻疼痛,改善下肢功能,治疗效果显著,且无明显不良反应。

电针调控背根神经节巨噬细胞浸润改善紫杉醇诱导神经病理痛的机制研究

目的:观察电针双侧足三里穴对紫杉醇诱导神经病理痛小鼠模型背根神经节(DRG)巨噬细胞标记物CD68、单核细胞趋化蛋白-1(MCP-1)、白细胞介素-1β(IL-1β)和诱导型一氧化氮合酶(iNOS)表达的影响,探讨电针调控紫杉醇诱导神经病理痛的机制。方法:选择成年C57小鼠30只,采用随机数字表法分为对照组、模型组、电针组,每组10只。模型组、电针组在第1、3、5、7天分别通过腹腔注射2 mg/kg紫杉醇进行造模;对照组注射相同体积的溶媒。造模结束后(第9天),电针组给予电针双侧足三里穴治疗,频率15 Hz,强度1 mA,30 min/次,1次/d,共7 d。采用“up and down”法测定足底机械阈值(PWMT);采用苏木精和伊红(HE)染色观察各组DRG组织的病理特征;采用免疫荧光染色法检测DRG组织MCP-1和CD68免疫阳性面积百分比;采用Western blot法检测DRG组织CD68、MCP-1、iNOS和IL-1β蛋白表达水平;采用透射电镜观察DRG组织巨噬细胞超微结构。结果:①PWMT:与对照组同一时间点比较,模型组第8、11、13、15天PWMT明显降低(P<0.05);与模型组同一时间点比较,电针组第11、13、15天PWMT明显升高,差异具有统计学意义(P<0.05)。②DRG组织病理学特征:炎症细胞形态为近圆形,颜色为蓝紫色,对照组有少量的炎症细胞,模型组聚集浸润状态的炎症细胞数量增加,电针组聚集浸润状态的炎症细胞数量较少。③DRG组织MCP-1、CD68免疫阳性面积百分比:与对照组比较,模型组DRG组织MCP-1、CD68免疫阳性面积百分比明显升高(P<0.05);与模型组比较,电针组DRG组织MCP-1、CD68免疫阳性面积百分比明显降低(P<0.05)。④DRG组织MCP-1、CD68、IL-1β和iNOS蛋白表达水平:与对照组比较,模型组DRG组织MCP-1、CD68、IL-1β和iNOS蛋白表达水平明显升高(P<0.05);与模型组比较,电针组DRG组织MCP-1、CD68、IL-1β和iNOS蛋白表达水平明显下降(P<0.05)。⑤DRG组织巨噬细胞超微结构:对照组DRG组织巨噬细胞形态呈近圆形,有圆钝形突起;模型组DRG组织巨噬细胞形态呈扁平状,有伪足,胞体比对照组大;电针组DRG组织巨噬细胞的形态呈近圆形,胞体比对照组大。结论:电针双侧足三里穴可改善紫杉醇诱导的神经病理痛小鼠疼痛程度,其机制可能是电针通过下调DRG中MCP-1表达,抑制巨噬细胞在DRG组织中的浸润,减少DRG组织中IL-1β、iNOS和CD68炎症因子释放,从而发挥镇痛作用。

电针对正常小鼠血清单胺类神经递质的影响

目的观察单次电针对小鼠血清中单胺类神经递质的影响。方法30只雄性C57BL/6J小鼠随机分为对照组和电针组,每组15只。电针组单次电针双侧足三里穴,对照组仅予电针组相同的束缚。采用高效液相色谱仪测定电针干预前及干预后1 h、2 h、4 h、8 h、24 h血清中去甲肾上腺素(norepinephrine,NE)、多巴胺(dopamine,DA)及其代谢产物3,4-二羟苯乙酸(3,4-dihydroxyphenylaceticacid,DOPAC)、5-羟色胺(5-hydroxytryptamine,5-HT)及其代谢产物5-羟基吲哚乙酸(5-hydroxyindole acetic acid,5-HIAA)的含量。结果干预前,两组血清各单胺类神经递质比较,差异无统计学意义(P>0.05)。两组干预后1 h、2 h、4 h和8 h血清NE含量较干预前降低(P<0.05,P<0.01);电针组干预后24 h血清NE含量显著低于对照组和干预前(P<0.01)。两组干预后血清DA含量和DOPAC/DA比值在各个时间点均降低(P<0.05,P<0.01),对照组干预后2 h血清DOPAC含量较干预前降低(P<0.05),电针组干预后4 h和8 h血清DOPAC含量较干预前升高(P<0.01);电针组干预后2 h、4 h、8 h和24 h血清DA含量高于对照组(P<0.05,P<0.01),DOPAC/DA比值低于对照组(P<0.01)。两组干预后各时间点血清5-HT含量均降低(P<0.01),两组血清5-HIAA在各个时间点差异无统计学意义(P>0.05);对照组干预后8 h和24 h血清5-HIAA/5-HT比值较干预前降低(P<0.05),电针组干预后1 h、4 h和24 h血清5-HIAA/5-HT比值较干预前升高(P<0.01);电针组干预后2 h和4 h血清5-HT含量低于对照组(P<0.05),干预后1 h、4 h、8 h和24 h血清5-HIAA/5-HT比值高于对照组(P<0.01,P<0.05)。结论单次电针可对生理状态下小鼠外周血血清单胺类神经递质的含量产生影响。

Adhesive ileus treated by electroacupuncture at Zhigou(TE 6) and Zusanli(ST 36):a ran-domized controlled study

Objective To assess the clinical efficacy on adhesive ileus treated by electroacupuncture(EA) at Zhigou(TE 6) and Zusanli(ST 36),and to explore the different effects of acupoint and non-acupoint. Methods Forty cases were randomized into an acupoint group

基于16S rDNA高通量测序研究电针对腹泻型肠易激综合征大鼠肠道菌群的调节作用

目的采用16S rDNA高通量测序技术,分析电针足三里对腹泻型肠易激综合征(irritable bowel syndrome-diarrhea,IBS-D)大鼠肠道菌群物种相对丰度和多样性的影响,探讨电针对IBS-D的作用机制。方法将SD大鼠随机分为空白组、模型组、电针组。采用束缚联合番泻叶煎剂灌胃法造模,电针组采用电针双侧足三里干预,空白组与模型组不做干预处理。干预结束后,检测大鼠内脏敏感性,并取大鼠粪便进行16S rDNA高通量测序技术检测大鼠肠道菌群。结果(1)与空白组比较,模型组大鼠腹壁回撤反射评分明显升高(P<0.05);经过电针治疗后,在20 mmHg压力下的腹壁回撤反射评分明显降低(P<0.05)。(2)与空白组相比,模型组OTU数目明显降低,电针组接近于空白组。(3)与空白组相比,模型组芽孢杆菌纲、乳杆菌属、真杆菌属等丰度明显降低,拟杆菌纲、梭菌纲、δ-变形杆菌纲、毛螺菌属等丰度明显升高,而电针组上述菌纲/属丰度均有所改善。(4)三组的Beta多样性分析结果显示,模型组与空白组的样本明显被区分开,而电针组则在两组之间。结论IBS-D与内脏高敏感性以及肠道菌群结构紊乱有关,电针能调节IBS-D大鼠内脏敏感性以及肠道菌群物种相对丰度,改善菌群多样性。

The Expression of Calcitonin Gene-related Peptide on the Neurons Associated Zusanli(ST 36)in Rats

Objective： To investigate the biochemical characteristic of the neurons associated Zusanli （ST 36） in the rat by using Alexa Fluor 594 conjugated cholera toxin subunit B （AF594-CTB） neural tracing and calcitonin gene-related peptide （CGRP） fluorescent immunohistochemical techniques. Methods： Four male Sprague Dawley rats were injected with AF594-CTB into the corresponding area of the Zusanli in the human body. After 3 surviving days, the rat＇s spinal cord and dorsal root ganglia （DRGs） at lumbar segments were dissected following perfusion with 4% paraformaldehyde, cut into sections, and then stained with CGRP- fluorescent immunohistochemical method. Results： AF594-CTB labeled sensory neurons were detected in the L3-L6 DRGs with high concentration in L4 DRG, and the labeled motor neurons located in the dorsolateral and intermediate regions of lamina IX from L3-L5 segments with high concentration at L4. Meanwhile, CGRP- positive neural labeling distributed symmetrically on both sides of DRGs, anterior and dorsal horns of spinal cord. In the AF594-CTB labeled neurons, 37% sensory neurons and 100% motor neurons expressed CGRP- positive. Conclusion： These findings present the morphological evidence to demonstrate that the sensory and motor neurons associated Zusanli in the rat distributed with segmental and regional patterns, and contained CGRP-expression.

Electroacupuncture at Feishu(BL13) and Zusanli(ST36) down-regulates the expression of orexins and their receptors in rats with chronic obstructive pulmonary disease

Inflammation and lung function decline are the main pathophysiological features of chronic obstructive pulmonary disease （COPD）. Acupuncture can improve lung function in patients with COPD, but the underlying mechanisms are not well understood. Orexins （OXs）, which are found in peripheral plasma, are neuropeptides that regulate respiration and their levels are related to COPD. Therefore, we hypothesized that acupuncture might alter OXs, reduce lung inflammation and improve lung function in COPD. METHODS： COPD was induced in rats by exposure to cigarette smoke for 8 weeks and injecting with lipopolysaccharide twice. Electroacupuncture （EA） was performed at Feishu （BL13） and Zusanli （ST36） for 30 min/d for 2 weeks. Rat lung function and morphology were assessed after EA. The levels of tumor necrosis factor-α （TNF-a） and intedeukin-1β （IL-16） in bronchoalveolar lavage fluid （BALF） and orexin A and B levels in the lung tissue were detected by enzyme-linked immunosorbent assay. OX receptor mRNA levels and immunopositive cells were assessed with real-time polymerase chain reaction and immunohistochemical methods, respectively. The relationships among lung function, cell factors, and OX levels were analyzed by Pearson correlation analyses. RESULTS： Compared with the control group, lung function was significantly decreased in the rats with COPD （P〈0.05）. There were obvious increases in TNF-α and IL-1β levels in BALF （P〈0.05 and P〈0.01, respectively）, orexin A level in lung tissue （P〈0.01; but not orexin B） and mRNA expressions of OX 1 type receptor （OXR1） and OX 2 type receptor （OXR2） in lung tissue （P〈0.05 and P〈0.01, respectively）; the integrative optical densities （IODs） of both receptors were greater in the COPD group （P〈0.05）. For rats with COPD subjected to EA, lung function was improved （P〈0.05）. There were notable decreases in TNF-a and IL-1β levels （P〈0.05 and 〈0.01, respectively） in BALF. Orexin A, but not orexin B, levels in lung tissue also decreased （P〈0.01）, as did mRNA expression of OXlR and OX2R in lung tissue （P〈0.05 and P〈0.01, respectively）. Receptor IODs were also reduced after EA treatment （P〈0.05）. Furthermore, orexin A levels and ratio of forced expiratory volume in 0.3 s to forced vital capacity were strongly negatively correlated （P〈0.01）, and orexin A was positively correlated with TNF-α and IL-1β （P〈0.001 and P〈0.05, respectively）.CONCLUSION： EA at Zusanli and Feishu improved lung function of rats with COPD and had an anti-inflammatory effect, which may be related to down-regulation of OXA and its receptors.