OBJECTIVE:To investigate the effect of moxibustion on synovitis and the autophagy of synoviocytes in rheumatoid arthritis(RA).METHODS:Forty Sprague-Dawley rats were randomly divided into a normal group,model group,mox...OBJECTIVE:To investigate the effect of moxibustion on synovitis and the autophagy of synoviocytes in rheumatoid arthritis(RA).METHODS:Forty Sprague-Dawley rats were randomly divided into a normal group,model group,moxibustion group,cigarette moxibustion group,and medicine group,with eight rats included in each group.The RA model was established by subcutaneous injection of complete Freund's adjuvant into the left posterior toe.Rats in the model group were not interfered with.In the moxibustion group,rats were treated by moxibustion,where a 1-cm diameter moxa stick was applied at the left Zusanli(ST 36)point.The distance of the moxa stick to the skin was 2 cm and moxibustion was completed for 20 min daily for 15 d total.In the cigarette moxibustion group,the moxa stick was replaced by a common cigarette.In the medicine group,rats were treated with a tripterygium glycoside suspension(8 mg/kg)once a day for 15 d total.In each group,the left hind limb toe volume was measured with a toe volume meter;the synovial cells were observed by hematoxylin and eosin staining;the interleukin(IL)-4,IL-6,IL-10,IL-1β,IL-23,IL-17,and tumor necrosis factor(TNF)-αlevels in serum were measured by enzyme-linked immunosorbent assay;the erythrocyte sedimentation rate(ESR)were detected by Westergren sedimentation rate testing;the C-reactive protein(CRP)and rheumatoid factor(RF)levels in serum were detected by rate nephelometry;the expression levels of ULK1,autophagy-associated protein(Atg)3,Atg5,and Atg12 messenger RNA(m RNA)in synovium were detected by real time-quantitative polymerase chain reaction(RT-q PCR);and the protein expression levels of phosphatidylinositol-3-kinase(PI3K),protein kinase B(Akt),mammalian target of rapamycin(mTOR),LC3-Ⅱ,beclin-1,phosphorylated-PI3K(p-PI3K),p-Akt,p-mTOR in synovium were detected by Western blotting.RESULTS:Among the RA model rats,joint swelling,an inflammatory reaction,and the proliferation of synovial tissue were obvious and the signal of the PI3K/Akt/mTOR pathway was active,while autophagy was inhibited.Moxibustion at Zusanli(ST36)or intragastric administration of Tripterygium wilfordii glycosides could alleviate the inflammatory reaction of RA rats;relieve the swelling of the toes;downregulate the levels of ESR,CRF,RF;lower the levels of IL-6,IL-1β,TNF-α,and IL-17;and increase the IL-4 and IL-10.At the same time,the m RNA expression levels of ULK1,Atg3,Atg5,and Atg12 and those of LC3-Ⅱand beclin-1 were increased,while the PI3K,Akt,mTOR,p-PI3K,p-Akt,p-mTOR were decreased.Cigarette moxibustion did not significantly reduce the swelling of the toe joint in RA rats,and was not as good as that of moxibustion or Tripterygium wilfordii polyglycosides in the effects of inflammation relief and the influences of the levels of ESR,CRF,RF.While cigarette moxibustion has a weak effect to affect the expression of corresponding molecules in autophages and the expression level of the autophagy biomaker in synovial tissue.Moxibustion and tripterygium glycosides can significantly reduce the joint swelling,relieve synovitis and synovial hyperplasia,and inhibit the PI3K/Akt/mTOR signaling pathway to increase autophagy in a manner superior to cigarette moxibustion.CONCLUSION:Moxibustion can limit the proliferation of synoviocytes in RA rats by inhibiting the PI3K/Akt/mTOR signaling pathway,promoting autophagy,effectively reducing synovitis,and alleviating joint swelling.展开更多
OBJECTIVE:To explore whether the paraventricular nucleus(PVN)participates in regulation of the antimyocardial ischemia-reperfusion injury(MIRI)effect of electroacupuncture(EA)and whether this is achieved through the P...OBJECTIVE:To explore whether the paraventricular nucleus(PVN)participates in regulation of the antimyocardial ischemia-reperfusion injury(MIRI)effect of electroacupuncture(EA)and whether this is achieved through the PVN-interposed nucleus(IN)neural pathway.METHODS:The modeling method of myocardial ischemia reperfusion injury was achieved by ligating the left anterior descending coronary artery in SpragueDawley rats.We used the Powerlab multi-channel physiological recorder system to record electrocardiograms and analyze the changes in ST segment displacement;2,3,5-Triphenyltetrazolium chloride staining was used to observe the percentage of myocardial infarction areas.Detecting cardiac troponin I(cTnI),lactate dehydrogenase(LDH)in serum was done with an enzyme-linked immunosorbent assay kit.Morphological changes in the myocardium were detected in each group with hematoxylin-eosin staining of paraffin sections.Detection of c-fos protein expression in the PVN of the hypothalamus was done with the immuneofluorescence method.The Plexon multi-channel acquisition system recorded PVN neuron discharges and local field potentials in each group of rats.Offline Sorter software was used for cluster analysis.Neuro Explorer software was used to perform autocorrelation,raster and frequency characteristics and spectral energy analysis of neuron signals in each group.RESULTS:Compared with the MIRI model group,the areas of myocardial infarction in the EA group were significantly reduced;the expression of cTnI,LDH in serum was decreased significantly.The firing frequency of pyramidal cells in the PVN was significantly increased and the spectrum energy map showed energy was reduced,c-fos expression in PVN was reduced,this indicated that neuronal activity in the PVN participates in the effect of EA improving myocardial injury.In addition,we used the kainic acid method to lesion the IN and observed that the effect of EA was weakened.For example,the area of myocardial infarction of lesion IN+EA group in rats was significantly increased compared with that resulting from EA group,the expression of cTnI,LDH in serum was significantly increased,the firing frequency of pyramidal cells in the PVN was significantly reduced.A spectral energy diagram shows that the energy after damage was higher than that of EA group.At the same time,the expression of c-fos in the PVN increased again.CONCLUSION:Our results indicated that the PVN-IN nerve pathway may participate as an effective pathway of EA to improve the effect of myocardial injury.展开更多
基金国家自然科学基金项目:8147378481774414+2 种基金安徽省高校自然科学研究重点项目:KJ 2017 A 298安徽省高校优秀青年人才支持计划重点项目:gxyq ZD 2016134安徽高校科研创新平台建设项目:2015 TD 033
文摘目的:观察胃俞募配穴针刺与胃俞、中脘单穴针刺对胃扩张受试者静息态脑功能局部一致性(regional homogeneity,ReHo)的影响及其与胃运动变化的相关性,探讨胃俞募配穴的中枢整合机制。方法:采用交叉试验设计,将24例健康受试者分3次先后纳入胃俞组、中脘组和胃俞配伍中脘组(配穴组),每次每组纳入8例,每组共纳入24例受试者,在水负荷胃扩张状态下分别接受胃俞穴、中脘穴和胃俞配伍中脘穴针刺,每次针刺前后分别进行胃电图检测和静息态脑功能磁共振成像(functional magnetic resonance imaging,fMRI)扫描,对针刺前后胃运动变化情况进行分析,对采集的fMRI图像均进行ReHo值计算,分析比较各组针刺前后和各组间ReHo变化;对针刺前后胃运动情况和ReHo变化进行相关性分析。结果:(1)针刺后,各组受试者胃电图振幅均明显低于针刺前(均P<0.01),胃电图频率均较治疗前差异无统计学意义(均P>0.05),胃俞组及中脘组胃电图振幅均高于配穴组(均P<0.05)。(2)与针刺前比较,胃俞募配穴针刺及胃俞、中脘单穴针刺引起了不同的大脑ReHo变化;胃俞募配穴针刺可引起右颞下回、左侧丘脑、楔前叶、后扣带回ReHo显著增高(均P<0.05),右侧颞极颞中回、距状沟、楔前叶ReHo明显减低(均P<0.05)。与单穴组比较,胃俞募配穴针刺均引起了后扣带回ReHo值增高和颞极ReHo值减低(均P<0.05)。(3)相关性分析显示,配穴组后扣带回、丘脑、楔前叶ReHo变化与胃运动振幅变化呈正相关,颞极ReHo变化与胃运动振幅变化呈负相关。结论:胃俞募配穴针刺及胃俞、中脘单穴针刺引起了不同的脑区ReHo变化。胃俞募配穴较单穴针刺可引起某些新的脑区ReHo变化,丘脑、后扣带回、楔前叶等脑区可能是胃俞募配穴针刺调节胃运动效应的重点整合脑区。胃俞募配穴针刺对胃运动的调节作用与丘脑、边缘系统和默认网络部分脑区密切相关。
基金Supported by the University Natural Science Research Program of Jiangsu:Study on the Mechanism Of Moxibustion Regulating Autophagy in RA synovial Fibroblasts Based on PI3K/Akt/mTOR Pathway(No.18KJB360005)the 973 Program of China:Effect and Biological Mechanism of Heat,Light and Smoking in Moxibustion(No.2015CB554504)+1 种基金funded by the Priority Academic Program Development of Jiangsu Higher Education Institutions(Integration of Chinese and Western Medicine)the Exploration and Scientific Research Project of Anhui University of Chinese Medicine:Clinical Application and Mechanism Study of Moxibustion(No.2016ts024)。
文摘OBJECTIVE:To investigate the effect of moxibustion on synovitis and the autophagy of synoviocytes in rheumatoid arthritis(RA).METHODS:Forty Sprague-Dawley rats were randomly divided into a normal group,model group,moxibustion group,cigarette moxibustion group,and medicine group,with eight rats included in each group.The RA model was established by subcutaneous injection of complete Freund's adjuvant into the left posterior toe.Rats in the model group were not interfered with.In the moxibustion group,rats were treated by moxibustion,where a 1-cm diameter moxa stick was applied at the left Zusanli(ST 36)point.The distance of the moxa stick to the skin was 2 cm and moxibustion was completed for 20 min daily for 15 d total.In the cigarette moxibustion group,the moxa stick was replaced by a common cigarette.In the medicine group,rats were treated with a tripterygium glycoside suspension(8 mg/kg)once a day for 15 d total.In each group,the left hind limb toe volume was measured with a toe volume meter;the synovial cells were observed by hematoxylin and eosin staining;the interleukin(IL)-4,IL-6,IL-10,IL-1β,IL-23,IL-17,and tumor necrosis factor(TNF)-αlevels in serum were measured by enzyme-linked immunosorbent assay;the erythrocyte sedimentation rate(ESR)were detected by Westergren sedimentation rate testing;the C-reactive protein(CRP)and rheumatoid factor(RF)levels in serum were detected by rate nephelometry;the expression levels of ULK1,autophagy-associated protein(Atg)3,Atg5,and Atg12 messenger RNA(m RNA)in synovium were detected by real time-quantitative polymerase chain reaction(RT-q PCR);and the protein expression levels of phosphatidylinositol-3-kinase(PI3K),protein kinase B(Akt),mammalian target of rapamycin(mTOR),LC3-Ⅱ,beclin-1,phosphorylated-PI3K(p-PI3K),p-Akt,p-mTOR in synovium were detected by Western blotting.RESULTS:Among the RA model rats,joint swelling,an inflammatory reaction,and the proliferation of synovial tissue were obvious and the signal of the PI3K/Akt/mTOR pathway was active,while autophagy was inhibited.Moxibustion at Zusanli(ST36)or intragastric administration of Tripterygium wilfordii glycosides could alleviate the inflammatory reaction of RA rats;relieve the swelling of the toes;downregulate the levels of ESR,CRF,RF;lower the levels of IL-6,IL-1β,TNF-α,and IL-17;and increase the IL-4 and IL-10.At the same time,the m RNA expression levels of ULK1,Atg3,Atg5,and Atg12 and those of LC3-Ⅱand beclin-1 were increased,while the PI3K,Akt,mTOR,p-PI3K,p-Akt,p-mTOR were decreased.Cigarette moxibustion did not significantly reduce the swelling of the toe joint in RA rats,and was not as good as that of moxibustion or Tripterygium wilfordii polyglycosides in the effects of inflammation relief and the influences of the levels of ESR,CRF,RF.While cigarette moxibustion has a weak effect to affect the expression of corresponding molecules in autophages and the expression level of the autophagy biomaker in synovial tissue.Moxibustion and tripterygium glycosides can significantly reduce the joint swelling,relieve synovitis and synovial hyperplasia,and inhibit the PI3K/Akt/mTOR signaling pathway to increase autophagy in a manner superior to cigarette moxibustion.CONCLUSION:Moxibustion can limit the proliferation of synoviocytes in RA rats by inhibiting the PI3K/Akt/mTOR signaling pathway,promoting autophagy,effectively reducing synovitis,and alleviating joint swelling.
基金Supported by National Natural Science Foundation of China:Mechanism of GABA/Glu Neural Circuit in Lateral HypothalamusParietal Nucleus in Alleviating Myocardial Ischemia-Reperfusion Injury by Acupuncture Preconditioning(82074536)Study on the Protective Effect of Acupuncture Pretreatment on Myocardial Ischemia-Reperfusion Injury Based on Hypothalamic-Cerebellar Neural Circuit(81774414)+2 种基金Mechanism of GABA Neural Circuit in the Paraventricular Nucleus of Hypothalamus and Ventrolateral Region of Medulla Oblongata in Alleviating Myocardial Ischemia-Reperfusion Injury Induced by Acupuncture Pretreatment(82104999)Natural Science Foundation of Anhui Province the Central Regulatory Mechanism of Acupuncture Regulating Cardiac Function(2108085Y30)Anhui Province University Outstanding Top Talent Cultivation Funding Project(gxgwfx2019025)
文摘OBJECTIVE:To explore whether the paraventricular nucleus(PVN)participates in regulation of the antimyocardial ischemia-reperfusion injury(MIRI)effect of electroacupuncture(EA)and whether this is achieved through the PVN-interposed nucleus(IN)neural pathway.METHODS:The modeling method of myocardial ischemia reperfusion injury was achieved by ligating the left anterior descending coronary artery in SpragueDawley rats.We used the Powerlab multi-channel physiological recorder system to record electrocardiograms and analyze the changes in ST segment displacement;2,3,5-Triphenyltetrazolium chloride staining was used to observe the percentage of myocardial infarction areas.Detecting cardiac troponin I(cTnI),lactate dehydrogenase(LDH)in serum was done with an enzyme-linked immunosorbent assay kit.Morphological changes in the myocardium were detected in each group with hematoxylin-eosin staining of paraffin sections.Detection of c-fos protein expression in the PVN of the hypothalamus was done with the immuneofluorescence method.The Plexon multi-channel acquisition system recorded PVN neuron discharges and local field potentials in each group of rats.Offline Sorter software was used for cluster analysis.Neuro Explorer software was used to perform autocorrelation,raster and frequency characteristics and spectral energy analysis of neuron signals in each group.RESULTS:Compared with the MIRI model group,the areas of myocardial infarction in the EA group were significantly reduced;the expression of cTnI,LDH in serum was decreased significantly.The firing frequency of pyramidal cells in the PVN was significantly increased and the spectrum energy map showed energy was reduced,c-fos expression in PVN was reduced,this indicated that neuronal activity in the PVN participates in the effect of EA improving myocardial injury.In addition,we used the kainic acid method to lesion the IN and observed that the effect of EA was weakened.For example,the area of myocardial infarction of lesion IN+EA group in rats was significantly increased compared with that resulting from EA group,the expression of cTnI,LDH in serum was significantly increased,the firing frequency of pyramidal cells in the PVN was significantly reduced.A spectral energy diagram shows that the energy after damage was higher than that of EA group.At the same time,the expression of c-fos in the PVN increased again.CONCLUSION:Our results indicated that the PVN-IN nerve pathway may participate as an effective pathway of EA to improve the effect of myocardial injury.
