ABSTRACT
BACKGROUND: Acupuncture preconditioning was able to reduce the extent of ischaemia reperfusion (I/R) injury. Previous studies have shown that electroacupuncture (EA) pretreatment at T4-T5 Jiaji (EX-B2) acupoints had cardioprotective effects against myocardial I/R injury. However, the molecular mechanism remains inconclusive. METHODS: Wistar rats were pretreated with electroacupuncture for 7 days at the Neiguan (PC6), T4-T5 Jiaji (EX-B2), Yanglingquan (GB34), and Quchi (LI11) acupoints, which belong to different meridians. Then, we investigated the genome-wide gene expression profiles of rats prestimulated at these acupoints after I/R injury. RESULTS: Our study revealed previously unknown cardioprotective roles of T4-T5 Jiaji (EX-B2) acupoints in the I/R progression. The extent of myocardial injury was significantly decreased in the Jiaji group compared with the I/R group. In addition, our data are among the first to link the EA preconditioning at Neiguan (PC6) acupoints and circadian rhythm in the I/R model. Also, for the first time, we explored the meridian and acupoint specificity involved in EA pretreatment at the heart meridian, in which Yanglingquan and Quchi acupoints were selected as the control group for heart-divergent-meridian and nonheart-meridian acupoints. CONCLUSIONS: The present study suggested that EA pretreatment at Jiaji alters genome-wide gene expression and protects the rat myocardium against I/R injury, which are most likely through neurohumoral regulation.
ABSTRACT
BACKGROUND: Stroke is one of the leading causes of death and disability worldwide. Scalp acupuncture and exercise therapy have been proven as two effective methods for the treatment of stroke. However, their combined action and mechanisms have not been fully elucidated. The present study aimed to investigate the protective effect of scalp acupuncture combined with exercise therapy on neurons in rats with ischemic brain injury. METHODS: 100 rats were randomly divided into 5 groups including sham group, model group, acupuncture group, rehabilitation group, and experimental group (scalp acupuncture combined with exercise therapy). Middle cerebral artery occlusion (MCAO) model in rats was established according to Longa modified suture method to mimic ischemic stroke. The modified Bedexer's neurological function score was used to evaluate the neurological deficits of rats and the brain infarct volume was measured using 2, 3, 5-triphenyl tetrazolium chloride monohydrate (TTC) staining. Moreover, the apoptosis in the hippocampus was detected by western blotting and terminal deoxynucleotidyl transferase dUTP nick end labeling (TUNEL) assay. The pro-inflammatory cytokines such as interleukin-1 beta (IL-1ß) and tumor necrosis factor-alpha (TNF-α), reactive oxygen species (ROS) and superoxide dismutase (SOD) were determined by corresponding kits. Immunohistochemistry or immunofluorescence was performed to detect the expression of brain-derived neurotrophic factor (BDNF), S100ß and glial fibrillary acidic protein (GFAP) in the hippocampi of rats. RESULTS: The neurological deficit score, the expression levels of apoptotic factors such as cleaved caspase-3 and Bax, and the TUNEL-positive cell rate of the experimental group were significantly lower than those of the acupuncture group and the rehabilitation group. However, apoptosis inhibitor Bcl-2 showed downregulated expression in the MCAO model rats but this trend was reverted by single and combinatorial treatments. In addition, the contents of TNF-α, IL-1ß and ROS in the acupuncture group and the rehabilitation group were significantly lower than those in the model group, but higher than the experimental group. While the opposite results were obtained in SOD activity. Furthermore, compared with the model group, the ratios of BDNF, S100ß, and GFAP-positive cells in the acupuncture, rehabilitation and experimental groups were significantly increased, and the highest ratios were recorded in the experimental group. CONCLUSIONS: This study demonstrated that scalp acupuncture combined with exercise therapy effectively counteracts ischemic brain injury via the downregulation of pro-inflammatory mediators and ROS, the increased production of the antioxidant enzyme SOD, neurotrophic factor BDNF and astrocyte activities.
Subject(s)
Acupuncture Points , Acupuncture Therapy , Apoptosis , Brain/pathology , Exercise Therapy , Infarction, Middle Cerebral Artery/prevention & control , Scalp , Animals , Apoptosis Regulatory Proteins/metabolism , Behavior, Animal , Brain/metabolism , Brain/physiopathology , Brain-Derived Neurotrophic Factor/metabolism , Cytokines/metabolism , Disease Models, Animal , Glial Fibrillary Acidic Protein/metabolism , Infarction, Middle Cerebral Artery/metabolism , Infarction, Middle Cerebral Artery/pathology , Infarction, Middle Cerebral Artery/psychology , Inflammation Mediators/metabolism , Male , Necrosis , Rats, Sprague-Dawley , Reactive Oxygen Species/metabolism , S100 Calcium Binding Protein beta Subunit/metabolism , Superoxide Dismutase/metabolismABSTRACT
OBJECTIVE: To observe the protective effect of acupuncture preconditioning at "Jiaji" (EX-B 2) on acute myocardial ischemia-reperfusion injury (MIRI) in the rats and explore the mechanism. METHODS: Fifty Wistar rats were randomly divided into a control group, a model group, a Jiaji group, a Neiguan group and a Quchi group, 10 rats in each one. In the Jiaji group, the Neiguan group and the Quchi group, electroacupuncture was given for preconditioning at "Jiaji" T4~T5 (EX-B 2), "Neiguan" (PC 6) and "Quchi" (LI 11) for 7 days before modeling. In the control group and the model group, the regular feeding was given, without any acupuncture. At the end of acupuncture, except the control group, ligating the left anterior descending coronary artery (LAD) was adopted to duplicate MIRI models in the rest groups. Electrocardio-gram (ECG) was monitored and ST-segment shift was analyzed. HE staining method was adopted to observe the morphology of cardiac tissue in the rats of the groups. The transmission electron microscope was used to observe myocardial cell ultrastructure. WST-1 method was used to determine the activity of serum superoxide dismutase (SOD), TBA method was used to determine the content of serum malondialehyde (MDA) and the real-time fluorescent quantitative PCR method to determine the expressions of Nrf 2 in ischemic myocardial tissue and downstream HO-1 gene. RESULTS: Compared with the control group, after LAD ligation, ST-segment was elevated and depressed in ECG apparently after reperfusion in the rest groups (all P<0.05). The ST-segment elevation in the Jiaji group and the Neiguan group was less than that in the model group (both P<0.05). Compared with the model group, SOD activity was increased apparently in the Jiaji group and the Neiguan group (both P<0.05), and MDA content was reduced apparently (both P<0.05). The effects in the Jiaji group were better than those in the Neiguan group (both P<0.05). Pathologically, "Jiajia" (EX-B 2) and "Neiguan" (PC 6) all improved the morphology of cardiac tissue and cell ultrastructure. The effects in the Jiaji group were much more significant and the improvements in the Quchi group were not apparent. Compared with the control group, the expressions of Nrf 2 and HO-1 gene in myocardial tissue were down-regulated in the model group (both P<0.05). Those were up-regulated apparently in the Jiaji group and the Neiguan group as compared with the model group (P<0.05, P<0.01). The up-regulation times of the expressions of Nrf 2 and HO-1 gene in the Jiajia group were the highest in comparison. CONCLUSIONS: Acupuncture preconditioning at "Jiaji" (EX-B 2) has the protective effect on cardiac ischemia and reperfusion damage, which is probably relevant with the up-regulation of Nrf 2-ARE pathway expression, the activation of endogenous anti-oxidative pathway, the improvement of oxygen free radical scavenging capacity and the alleviation of lipid peroxide damage.
Subject(s)
Acupuncture Therapy , Ischemic Preconditioning, Myocardial/methods , Myocardial Reperfusion Injury/prevention & control , Oxidative Stress , Acupuncture Points , Animals , Electroacupuncture , Myocardial Reperfusion Injury/metabolism , Random Allocation , Rats , Rats, WistarABSTRACT
OBJECTIVE: To investigate the effect of "Jiaji"(EX-B 2)-electroacupuncture(EA) preconditioning on the myocardium and expression of cytochrome P 450 signaling pathways of myocardial cells in myocardial ischemia-reperfusion injury(MI/RI) rats. METHODS: Sixty male Wistar rats were randomly divided into sham operation, model, Jiaji, Neiguan(PC 6), Yanglingquan(GB 34) and Quchi(LI 11) groups (n=10 in each group). MI/RI model was established by occlusion of the descen-ding anterior branch of the left coronary artery. EA preconditioning was respectively applied to EX-B 2, PC 6, GB 34 and LI 11 for 30 min, once daily for 7 days. The ECG-ST segment was detected before and after the modeling. The contents of serum creatine kinase-MB(CK-MB) and lactate dehydrogenase(LDH) were detected by ELISA and microplate method separately. The expression levels of myocardial CYP 450 related genes cyp 1 a 1, cyp 2 b 2, cyp 2 c 11, cyp 4 a 2 were detected by Real-time PCR. RESULTS: Thirty minutes after ligation the changes of ECG-ST segment displacement of the Jiaji group and Neiguan group were signifi-cantly lower than that of the model group(P<0.01), and the change was significantly lower in the Neiguan group than in the model group(P<0.01) 30 minutes after reperfusion. Compared with the sham operation group, the activities of serum CK-MB and LDH were significantly increased in the model group(P<0.01). Compared with the model group, the activities of serum CK-MB and LDH were significantly decreased in the Jiaji, Neiguan and Yanglingquan groups (P<0.01). Compared with the Jiaji group, the activity of serum LDH was significantly increased in the Quchi group, while the activities of serum CK-MB were significantly increased in the Quchi and Yanglingquan groups (P<0.05). Compared with the sham operation group, the expression levels of myocardial cyp 1 a 1 and cyp 2 b 2 mRNAs were significantly higher in the model group(P<0.01), while the expression of cyp 2 c 11 mRNA was lower(P<0.01). Compared with the model group, the expression levels of cyp 1 a 1 and cyp 4 a 2 mRNAs were lower in the Jiaji, Neiguan and Yanglingquan groups (P<0.01), while the expressions of cyp 2 b 2 and cyp 2 c 11 mRNAs were higher(P<0.01). Compared with the Jiaji group, the expression of cyp 1 a 1 mRNA was lower in the Neiguan group, while the expression of cyp 2 b 2 mRNA was higher(P<0.05). CONCLUSIONS: EA preconditioning at Jiaji(EX-B 2) can reduce serum myocardial enzymes of myocardial ischemia-reperfusion rats, lower the ST segment of electrocardiogram, up-regulate myocardial CYP 450 epoxygenase cyp 2 b 2 and cyp 2 c 11 mRNAs expression, down-regulate ω-hydroxylase cyp 1 a 1 and cyp 4 a 2 mRNAs expression, thus play a role in myocardial protection.