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.

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.

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.

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.

TREATMENT OF OBSTINATE PEPTIC ULCER AND CHRONIC GASTRITIS WITH A METAL RING EMBEDDED IN ZUSANLI ACUPOINT

We had treated 102 cases of obstinate peptic ulcer and chronic gastritis with ametal ring embedded in Zusanli acupoint since October 1989 to October 1992,and got satisfactorytherapeutic effects.The total effective rate is 96.08%,the cure rate of peptic ulcer is 87.32%,therecurrence rate is significantly lower than that of control group(P【0.001),and it has no side-effect.

CONSTIPATION TREATED BY AURICULAR ACUPOINT PRESSING COMBINED WITH ACUPUNCTURE AT ZUSANLI POINT

This paper reports that 65 cases with functional constipation were treated by putting one Wang Bu Liu Xing (semen vaccariae) seed on each acupoint, Intestinum crassum, Intestinum tenue,Sympathicum, Lung, Spleen, Subcortex, of auricula and pressing, associated with acupuncture at Zusanli point on both sides. The patients were instructed to press the stuck seed twice a day for 3 to 5 minutes each time. Auricular acupoints of both sides were used alternately every 2 days. The electric acupuncture was applied at Zusanli points about 30 minutes each time every other day. Ten times (i. e. 20 days) make one course of treatment and the routine treatment consists of 1 to 2 courses. The results showed that all the patients were cured, patients with Shi Mi (the constipation of excess type) were cured after one course of treatrnent while Xu Mi (the constipation of deficiency type) were cured after two courses. The study suggests that this kind of treatment is an ideal therapy for functional constipation. The pathogenesis and mechanism of treatment of auricular acupuncture pressing associated with acupuncture at Zusanli point for constipation are discussed.

A COMPARATIVE OBSERVATION ON THE EFFECTS OF ACUPUNCTURING RENZHONG AND ZUSANLI ON THE NEURONAL ACTIVITY OF LOCUS COERULEUS IN RATS

In present work,the effects of acupuncturing Renzhong（GV 26）and Zusanli（ST 36）on the neuronal activity of Locus Coeruleus（LC）in rats were observed and compared.The resultsindicated that the central mechanism of promoting blood pressure（BP）by acupuncturing Renzhongwas related to the activity of LC;the influence of acupuncturing Renzhong on neuronal activity of LC was more obvious than that of acupuncturing Zusanli;and there was relative speciality between the acupoints.

Expression of ceramide galactosy transferase in sciatic nerve of rats with diabetic peripheral neuropathy after electroacupuncture at Zusanli and Shenshu points

BACKGROUND： Ceramide galactosyltransferase （CGT） protein and mRNA expression defect can cause the abnormal morphology and slowing conduction velocity of peripheral nerve. Morphologic change and functional disorder of myelin sheath and axon appear when diabetic peripheral neuropathy （DPN） occurs. Whether electroacupuncture at Zusanfi（ST 36） and Shenshu（BL 32） points can enhance the expression of CGT protein and mRNA in the DPN tissue？ OBJECTIVE： To observe the effect of electroacupuncture at Zusanfi and Shenshu points on motor, sensory conduction velocity and CGT mRNA and its protein expression of sciatic nerve in rats with DPN. DESIGN： A randomized and controlled animal experiment SETTING ： Department of Neurology and Central Laboratory, Yueyang Hospital of Traditional Chinese ＆ Western Medicine, Shanghai University of Traditional Chinese Medicine. MATERIALS ： Totally 60 healthy male Wistar rats of clean grade, aged 4 month, with body mass of 200 to 220 g, were enrolled in this study. Streptozotocin （STZ, Sigma Company of USA, Batch No. S-0130）. METHODS： This study was carried out in the Animal Experimental Center and Central Laboratory, Yueyang Hospital of Traditional Chinese ＆ Western Medicine during February 2005 to March 2006. （1） Fifteen rats were randomly chosen,serving as normal group.AU the other rats were intraperitoneally injected once with STZ to develop experimental diabetic rat models. If fasting blood glucose was ≥ 15 mmol/L,sensory nerve and motor nerve conduction velocity of sciatic nerve was obviously slowed, tail-swaying temperature threshold was increased and myelinated nerve fiber of sciatic nerve changed, DPN models were successful. The successful model rats were randomly assigned into 3 groups： model group, control group（electroacupuncture at non-meridian-non-acupoint）and electroacupuncture group [electroacupuncture at Zusan/i and Shenshu points], with 15 rats each. The rats in the normal group and model group were untouched. In the electroacupuncture group （electroacupuncture at Zusanfi and Shenshu points）, Shenshu point （double） and Zusanfi point （double） were chosen referencing to The Atlas of the Rat＇s Acupoints. G6805- Ⅱ electric acupuncture apparatus was used, and current intensity was controlled at 20 min/time, once every other day, 12 times within 24 days. In the control group, the tip of rat-tail was stimulated, and the concrete procedures were the same as in the electroacupuncture at Zusanfi and Shenshu points. （2） Motor nerve conduction velocity and sensory nerve conduction velocity of rats were detected with neuroelectrophysiology detector in the end of the treatment, and the expressions of CGT protein and mRNA of sciatic nerve were detected with immunohistochemical method and fluorescent quantitative PCR technique. MAIN OUTCOME MEASURES： （1） Motor and sensory nerve conduction velocity. （2） The expression of CGT protein and its mRNA. RESULTS： All the 60 rats entered the stage of result analysis. （1） Comparison of motor and sensory nerve conduction velocity of rats after electroacupuncture： Motor nerve conduction velocity of rats in the model group[（31.37±3.69） m/s], control group [（32.74±5.42） m/s] and electroacupuncture group [（41.30 ±1.15） m/s] was significantly lower than that in the normal group [（41.30±1.15） m/s, P 〈 0.01]; The sensory nerve conduction velocity of rats in the model group[（18.17±9.54） m/s], control group [（21.39±5.61） m/s]and electroacupuncture group [（35.81 ±4.59） m/s] was significantly lower than that in the normal group [（46.38± 6.32） m/s,P 〈 0.01]; The motor and sensory nerve conduction velocity of electroacupuncture group was significantly higher than that in the model group [（38.04±2.01） m/s vs. （32.74±5.42） m/s,（35.81±4.59） m/s vs. （21.39±5.61） m/s,P 〈 0.01]. （2） Comparison of the expression of CGT protein of sciatic nerve of rats： The number of CGT positive cells of sciatic nerve in model group, control group or electroacupuncture group was significantly smaller than that in normal group [（9 770.33±1 461.73）, （10 588.13±1119.52）, （27 518.27± 9 078.29）, （37 769.67±4 021.81）/μm^2,P 〈 0.01]; The number of CGT positive cells of the sciatic nerve in the electroacupuncture group was significantly larger than that in the model group and control group （P 〈 0.01）. The number of CGT positive cells of sciatic nerve was close between control group and electroacupuncture group （P 〉 0.05）. （3） Comparison of CGT mRNA expression of sciatic nerve of rats： Ct value of CGT mRNA of sciatic nerve of rats in the model group,control group and electroacupuncture group was significantly higher than that in the normal group （13.75±2.60,14.81±2.80,11.67±1.75,9.30±0.98, P 〈 0.01 ）; Ct value of CGT mRNA of sciatic nerve of rats in the electroacupuncture group was significantly lower than that in the model group and control group （P 〈 0.01）, and that was close between electroacupuncture group and control group （P 〉 0.05）. CONCLUSION ： Electroacupuncture at Zusanfi and Shenshu points can increase motor and sensory nerve conduction velocity of rats with DPN. It might be associated with up-regulating the expression of CGT mRNA and its protein.

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.

Effects of moxibustion at"Zusanli"(ST 36)on oxidative stress and intestinal flora in aging rats

Objective To observe the effects of moxibustion at"Zusanli"(ST 36)on oxidative stress and intestinal flora in subacute aging rats,and to explore the possible mechanism of moxibustion in delaying aging.Methods Thirty SD rats were randomly divided into a blank group,a model group and a Zusanli group,with 10 rats in each group.Subacute aging model was established by intraperitoneal injection of D-galactose at dosage of 500 mg/kg in the model group and the Zusanli group,once a day for 42 days.In the Zusanli group,moxibustion was applied at bilateral"Zusanli"(ST 36),once a day,3 moxa cones at each acupoint,for consecutive 28 days.After intervention,the serum levels of superoxide dismutase(SOD)and malondialdehyde(MDA)were detected by ELISA;the intestinal flora was detected by 16S rRNA sequencing technique in each group.

附子理中汤联合足三里穴位注射配合循经热熨法对腹腔镜胃癌术后(脾胃虚寒证)患者胃肠激素、炎症反应、营养参数和血清MIP-3α、Stathmin的影响

目的:观察附子理中汤联合足三里穴位注射配合循经热熨法对腹腔镜胃癌术后(脾胃虚寒证)患者胃肠激素、炎症反应、营养参数和血清微管解聚蛋白(Stathmin)、巨噬细胞炎性蛋白-3α(MIP-3α)的影响。方法:选择2021年1月至2023年3月该院收治的胃癌患者126例,分组方式为随机数字表法。对照组42例患者(脱落2例,最终纳入40例)给予常规西医治疗,观察1组42例患者(脱落3例,最终纳入39例)在对照组的基础上给予足三里穴位注射配合循经热熨法治疗,观察2组42例患者(脱落2例,最终纳入40例)在观察1组的基础上给予附子理中汤治疗。比较三组患者的临床疗效,记录肛门首次排气时间、首次经口进食时间和首次排便时间;术后1、15 d测定三组患者营养参数(血清血红蛋白、转铁蛋白和白蛋白),检测炎症反应指标[血红素加氧酶-1(HO-1)、缺氧诱导因子-1α(HIF-1α)和白细胞介素6(IL-6)]、胃肠激素指标[胃泌素17(G-17)、胃动素(MTL)]及MIP-3α、Stathmin水平。结果:观察1组、观察2组患者的总有效率分别为94.87%(37/39)、97.50%(39/40),高于对照组的75.00%(30/40),差异有统计学意义(P<0.05);而观察2组与观察1组患者总有效率的差异无统计学意义(P>0.05)。观察2组患者肛门首次排气时间、首次经口进食时间和首次排便时间短于对照组、观察1组,且观察1组患者短于对照组,差异均有统计学意义(P<0.05)。术后15 d,观察2组患者转铁蛋白、血红蛋白和白蛋白水平高于对照组、观察1组,且观察1组患者高于对照组;观察2组患者HO-1、HIF-1α和IL-6水平低于对照组、观察1组,且观察1组患者低于对照组;观察2组患者G-17、MTL水平高于对照组、观察1组,且观察1组患者高于对照组;观察2组患者Stathmin、MIP-3α水平低于对照组、观察1组,且观察1组患者低于对照组,上述差异均有统计学意义(P<0.05)。结论:附子理中汤联合足三里穴位注射配合循经热熨法治疗腹腔镜胃癌术后患者,可减少炎症反应,调节胃肠激素和营养参数,降低Stathmin、MIP-3α水平,促进疾病康复,提高临床疗效。

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.

针刺合谷、内关、足三里联合药物治疗急性胆囊炎的临床研究

目的分析针刺合谷、内关、足三里联合药物治疗急性胆囊炎(AC)的效果。方法选择2021年3月至2022年3月郏县中医院收治的AC患者62例,按照随机数字表法分为A组(常规药物治疗)和B组(常规药物治疗+针刺合谷、内关、足三里),对比两组治疗有效率、并发症发生率、临床症状评分、胆囊指标、炎性因子指标、疼痛程度、生活质量评分。结果B组治疗有效率高于A组,并发症发生率低于A组,差异具有统计学意义(P<0.05);治疗后B组临床症状评分(厌食、黄疸、口苦、发热、便秘、恶心、腹痛)、胆囊指标(胆囊横径、胆囊壁厚度)、炎性因子指标[白细胞计数(WBC)、高敏C反应蛋白(hs-CRP)、降钙素原(PCT)、白介素-6(IL-6)]、VAS、PPI、PRI评分低于A组,差异具有统计学意义(P<0.05);B组生活质量评分(躯体疼痛、社会功能、情感职能、活力、生理职能、生理机能、精神健康)高于A组,差异具有统计学意义(P<0.05)。结论AC患者在常规服用药物治疗的基础上联合针刺合谷、内关、足三里,能更好地改善患者临床症状、疼痛感,降低炎症指标、胆囊指标,在一定程度上降低并发症风险,提高患者生活质量。

坐骨神经胞外核苷酸酶在针刺缓解大鼠踝关节炎性痛中的作用

目的 探讨坐骨神经胞外核苷酸酶[CD39(NTPDase1)]在针刺缓解大鼠踝关节炎性痛中的作用。方法 将54只雄性大鼠随机分为9组,即空白组、踝关节炎性痛模型组、针刺组、坐骨神经腺苷三磷酸(ATP)水解酶抑制(ARL67156注射)+针刺组、坐骨神经CD39上调(三磷酸腺苷双磷酸酶注射)组、坐骨神经CD39特异性抑制(噻氯匹啶注射)+针刺组、坐骨神经CD73特异性抑制(α,β-亚甲基腺苷5′-二磷酸钠盐注射)+针刺组、穴区CD39上调(三磷酸腺苷双磷酸酶注射)组、穴区CD39特异性抑制(噻氯匹啶注射)+针刺组,每组6只。除空白组外,其余各组大鼠均采用注射完全弗氏佐剂建立佐剂性关节炎疼痛模型;造模48 h后,需针刺组大鼠针刺患侧“足三里”穴20 min,需注射药物组于针刺前20 min进行局部相关试剂注射。以大鼠患侧足底机械痛阈值和热痛阈值作为疼痛评价指标;采用实时荧光定量逆转录聚合酶链式反应(RT-qPCR)法和Western blot法检测坐骨神经以及“足三里”穴胞外核苷酸酶的表达。结果 (1)调节坐骨神经ATP水解酶活性:与空白组比较,踝关节炎性痛模型组和坐骨神经ATP水解酶抑制+针刺组热痛阈值和机械痛阈值明显下降(P<0.05);与踝关节炎性痛模型组比较,针刺组热痛阈值和机械痛阈值明显上升(P<0.05)。(2)调节坐骨神经CD39活性:与空白组比较,坐骨神经CD39特异性抑制+针刺组热痛阈值和机械痛阈值明显下降(P<0.05);与踝关节炎性痛模型组比较,针刺组和坐骨神经CD39上调组热痛阈值和机械痛阈值明显上升(P<0.05);与针刺组比较,坐骨神经CD39特异性抑制+针刺组热痛阈值和机械痛阈值明显下降(P<0.05)。(3)调节穴区CD39活性:与空白组比较,穴区CD39特异性抑制+针刺组热痛阈值和机械痛阈值明显下降(P<0.05);与踝关节炎性痛模型组比较,针刺组和穴区CD39上调组热痛阈值和机械痛阈值明显上升(P<0.05);与针刺组比较,穴区CD39特异性抑制+针刺组热痛阈值和机械痛阈值明显下降(P<0.05)。(4)调节坐骨神经CD73活性:与空白组比较,坐骨神经CD73特异性抑制+针刺组热痛阈值和机械痛阈值明显下降(P<0.05);与踝关节炎性痛模型组比较,针刺组热痛阈值和机械痛阈值明显上升(P<0.05)。(5)针刺对坐骨神经以及穴区CD39表达的影响:Western blot法检测结果显示,与空白组比较,踝关节炎性痛模型组坐骨神经CD39的表达显著增加(P<0.05);与踝关节炎性痛模型组比较,针刺组CD39表达显著下调(P<0.05)。RT-qPCR法检测结果显示,与踝关节炎性痛模型组比较,针刺组CD39 mRNA表达水平明显上调(P<0.05)。结论 坐骨神经局部CD39参与了针刺缓解大鼠踝关节炎性痛的机制,这可能与CD73联合促进胞外ATP向腺苷转化有关。

针刺足三里、内关穴对老年髋关节置换术后镇痛疗效及对血清炎症因子的影响

目的探讨针刺足三里、内关穴对老年髋关节置换术后镇痛疗效及对血清炎症因子的影响。方法选择196例行全髋关节置换术的老年患者作为研究对象,采用随机数字法平均分为观察组与对照组:对照组术后给予常规对症止痛及冰袋冷敷治疗,观察组术后6 h在对照组基础上给予足三里、内关穴针刺治疗。于术后6 h、24 h、48 h、72 h及第5天采用疼痛视觉模拟评分(Visual analogue scale,VAS)评价患者疼痛状态,于术后6 h、48 h及72 h评价术后胀气情况;于术后6 h、24 h、48 h及72 h检测血清炎症因子:前列腺素E2(Prostaglandin E2,PGE2)、白细胞介素-1β(Interleukin-1β,IL-1β)、肿瘤坏死因子-α(Tumor necrosis factor-α,TNF-α);于髋关节置换术前、术后72 h、出院时采用Harris髋关节评分标准评价髋关节功能。结果两组患者术后24 h VAS评分最高,随后逐渐降低(P<0.05);术后48 h观察组VAS评分降至术后6 h水平,而对照组仍高于术后6 h水平(P<0.05);术后24 h、48 h、72 h及第5天观察组VAS评分均低于对照组(P<0.05)。术后两组患者腹胀评分均逐渐降低(P<0.05);术后48 h、72 h观察组腹胀评分低于对照组(P<0.05)。观察组术后肠鸣音出现及恢复正常时间、首次排气及排便时间均较对照组短(P<0.05)。两组患者术后24 h PGE2、IL-1β、TNF-α水平最高,术后72 h逐渐降低(P<0.05);术后5 d观察组PGE2、IL-1β、TNF-α水平降至术后6 h水平,而对照组仍高于术后6 h水平(P<0.05);术后24 h、72 h及第5天观察组PGE2、IL-1β、TNF-α水平均低于对照组(P<0.05)。两组术后Harris髋关节评分均逐渐升高(P<0.05);术后72 h及出院时观察组Harris髋关节评分高于对照组(P<0.05)。结论针刺足三里、内关穴可以更好地降低老年髋关节置换术后疼痛程度,缩短疼痛时间,同时更好地促进胃肠道功能恢复,其机制可能与降低PGE2、IL-1β、TNF-α等炎症因子相关。

基于miRNA-29c/白血病抑制因子轴探讨针刺调节脾虚大鼠肠道炎症机制研究

目的观察针刺调节miRNA-29c/白血病抑制因子(leukemia inhibitory factor,LIF)轴改善脾虚大鼠结肠炎症机制,探讨“健脾益气”针刺疗法改善脾虚便溏症状现代作用机理。方法将60只SD雄性大鼠以随机数字表法分为正常组、模型组、非经非穴组、针刺组,每组15只。除正常组外,其余3组采用劳倦过度和不规律饮食复合方法建立脾虚证大鼠模型。造模成功后,针刺组电针“足三里”穴及“脾俞”穴干预治疗,非经非穴组给予电针尾尖作为对照干预,每日1次/20 min,共治疗14 d。治疗结束后,取大鼠结肠组织及全血。qPCR检测大鼠结肠组织miRNA-29/LIF轴相关基因的表达;Western blot检测大鼠白介素-6(interleukin6,IL-6)、肿瘤坏死因子α(tumornecrosisfactor-α,TNF-α)的表达;ELISA检测结肠组织中白介素-1β(interleukin1β,IL-1β)、白介素-18(interleukin18,IL-18)的含量;HE染色法观察大鼠结肠组织病理形态变化;TUNEL染色观察各组结肠组织凋亡情况;流式细胞术观察外周血单个核细胞(PBMC)凋亡状况。结果与正常组相比,模型组大鼠结肠组织LIF的表达水平明显下降(P<0.05),miRNA-29、IL-1β、IL-6、白介素-8(interleukin18,IL-8)、TNF-αmRNA表达以及IL-6、TNF-α蛋白表达水平有所升高(P<0.05);与模型组相比,针刺组大鼠结肠组织LIF表达水平明显升高(P<0.05),miRNA-29c、IL-1β、IL-6、IL-8、TNF-αmRNA表达以及IL-6、TNF-α蛋白表达水平有所降低(P<0.05),非经非穴组差异无统计学意义(P>0.05);与正常组相比,模型组大鼠结肠组织中促炎因子IL-1β、IL-18表达水平明显升高(P<0.05),与模型组相比,针刺组大鼠结肠组织中促炎因子IL-1β、IL-18的表达水平有所降低(P<0.05)。另外HE染色结果显示,针刺治疗可有效减轻结肠黏膜组织中炎症细胞浸润和杯状细胞的损失;TUNEL染色结果显示,针刺可以抑制miRNA-29c/LIF轴,改善脾虚大鼠结肠炎症,抑制结肠黏膜细胞凋亡;流式细胞术结果显示与正常组相比,模型组、非经非穴组大鼠PBMC凋亡率明显升高(P<0.05),与模型组相比,针刺组大鼠PBMC细胞凋亡率有所降低(P<0.05)。结论电针“足三里”“脾俞”穴可以调节miRNA-29c/LIF轴,抑制肠道炎症和促炎症介质的表达,进而抑制血液、结肠黏膜细胞凋亡,提高机体免疫力,从而改善脾虚便溏症状。

针刺足三里、阳陵泉穴对坐骨神经损伤Wistar大鼠NGF、BDNF表达的影响

目的:比较针刺足三里穴、阳陵泉穴对Wistar大鼠坐骨神经损伤(SNI)的功能恢复作用差异。方法:将48只SPF级雄性Wistar大鼠随机分为四组:假手术组、模型组、阳陵泉组、足三里组,每组12只,并采用物理损伤法制备大鼠坐骨神经损伤模型。造模结束24 h后,根据治疗方案,阳陵泉组和足三里组分别针刺大鼠双侧阳陵泉穴及足三里穴,直刺进针5 mm并留针15 min,每日1次,连续针刺治疗28 d。假手术组仅接受外科手术暴露坐骨神经而不进行挤压,不进行任何针刺治疗处理。观察针刺治疗后大鼠坐骨神经功能指数(SFI)的变化、受损坐骨神经组织病理学改变、免疫荧光法和Western blot检测受损坐骨神经组织中神经生长因子(NGF)和脑源性神经营养因子(BDNF)蛋白的表达变化、Hoechst 33258染色观察雪旺细胞的参与情况。结果:与假手术组相比,模型组大鼠行走印迹明显减轻,SFI评分显著下降(P<0.01);HE染色结果显示模型组坐骨神经组织纤维结构紊乱,排列混乱,呈不规则形式,且周围雪旺细胞分布增多、变圆,部分变形呈空泡状变性。免疫荧光与蛋白质印迹分析显示模型组坐骨神经中NGF、BDNF蛋白阳性表达显著增加(P<0.01);Hoechst 33258染色结果显示模型组细胞核数量明显增加(P<0.01)。与模型组比较,足三里组、阳陵泉组大鼠行走印迹明显加深,SFI评分显著上升(P<0.01),足三里组较阳陵泉组各指标改善更显著(P<0.01);HE染色结果显示足三里、阳陵泉组大鼠坐骨神经组织纤维整体排列序列性尚整齐,仅有部分髓鞘脱落,轴索水肿减轻,雪旺细胞增殖分化,炎性细胞浸润明显减少,并观察到神经纤维出现新的生长,且足三里组病理变化改善程度优于阳陵泉组。坐骨神经中NGF、BDNF蛋白阳性表达明显上调(P<0.01),足三里组NGF、BDNF蛋白表达较阳陵泉组上调更显著(P<0.01);细胞核数量增加(P<0.01)。结论:针刺足三里穴、阳陵泉穴有助于促进受损坐骨神经的结构和功能恢复,并显著上调Wistar大鼠坐骨神经损伤处NGF、BDNF蛋白的表达,且针刺足三里穴对受损坐骨神经损伤的改善效果更好,从而促进受损神经再生与修复。

曲安奈德足三里穴位注射治疗非过敏性鼻炎的临床观察

目的观察曲安奈德注射液足三里穴位注射治疗非过敏性鼻炎的临床疗效并探讨其作用机制。方法随机选取非过敏性鼻炎(non-allergic rhinitis,NAR)与过敏性鼻炎(allergic rhinitis,AR)门诊患者各50例,均予以双侧足三里穴位注射曲安奈德注射液治疗,每周1次,连续注射4周。治疗前后检测患者Th1、Th2相关因子水平;观察两组治疗4周时、3月后以及1年后随访期间的鼻部症状总评分(TNSS)变化,比较分析两组近期疗效和远期疗效。结果NAR组与AR组在治疗4周结束时、疗后3月及1年随访期间的TNSS均比治疗前明显下降(P<0.05)。NAR组与AR组患者治疗4周后的血清白介素-2(IL-2)、干扰素-γ(INF-γ)、肿瘤坏死因子(TNF)水平均比治疗前上升(P<0.05),白介素-4(IL-4)则较治疗前显著下降(P<0.05)。治疗4周后NAR组有效率84.0%,AR组有效率92.0%。随访期间,疗后3月NAR组有效率72.0%,AR组有效率64.0%,两组疗效差异无统计学意义(P>0.05);1年后,NAR组总有效率54.0%,AR组总有效率30.0%,NAR组总有效率显著高于AR组(P<0.05)。结论足三里穴位注射治疗NAR远期疗效更佳,对脾气虚弱和肺气虚寒型的治疗有效率更高。其机制可能与提升Th1细胞因子的分泌并抑制IL-4的产生,从而调节Th1/Th2的稳定状态相关。