OBJECTIVE: To investigate the effects on the motor function, cortex blood flow perfusion, microglial cells, and the contents of serum inflammatory factors, i.e. interleukin-1β (IL-1β), transforming growth factor-β (TGF-β), and interleukin-10 (IL-10) after electroacupuncture (EA) preconditioning at \"Baihui\" (GV20) and \"Dazhui\" (GV14) in the mice with ischemic stroke, so as to explore the mechanism of EA preconditioning for improving motor function after ischemic stroke.
METHODS: C57BL/6 mice were randomly divided into sham-operation group, model group, and EA preconditioning group (EA group), with 15 mice in each group. A photothrombotic method was used to induce the model of unilateral ischemic stroke and motor impairment. The mice in the EA group received EA preconditioning, 20 min each time, once daily for 7 consecutive days before modeling. The motor function of mice was evaluated by the grid-walking test and cylinder test before and after modeling. Laser speckle blood flow video monitoring system was employed to assess the cerebral blood flow perfusion in the primary motor cortex of mice. The contents of IL-1β, TGF-β, and IL-10 in the serum were measured by ELISA, and the expressions of microglial cell and M2 subtype cell marker in the primary motor cortex were detected using immunofluorescence staining.
RESULTS: After modeling, compared with the sham-operation group, the grid error rate and the dragging rate of the affected limb were increased (P<0.01)；the utilization rate of the affected limb and percentage of the blood perfusion in the affected cortex to healthy side were decreased (P<0.01)；the contents of serum IL-1β, TGF-β, and IL-10 were increased (P<0.01, P<0.05)；and the microglia in the primary motor cortex on the affected side showed ameboid, the fluorescence intensity of ionized calcium-binding adapter molecule 1 (IBA1) and CD206 was increased (P<0.01) in the model group. In the EA group, when compared with the model group, the grid error rate and the dragging rate of affected limb were decreased (P<0.01)；the utilization rate of affected limb and the percentage of blood perfusion were increased (P<0.05)；the content of serum IL-1β was decreased (P<0.01), while the contents of TGF-β and IL-10 were increased (P<0.01)；and the microglia in the primary motor cortex on the affected side got more round and were distributed more densely, the fluorescence intensity of IBA1 and CD206 was increased (P<0.01).
CONCLUSIONS: Electroacupuncture preconditioning at \"GV20\" and \"GV14\" can up-regulate the expression of microglial cells, especially the M2 subtype cell marker, and increase the contents of the anti-inflammatory factors and decrease that of the pro-inflammatory factors in the serum, thereby alleviate the inflammatory reaction.
目的: 观察电针预处理“百会”“大椎”对缺血性中风小鼠运动功能、脑皮层血流灌注、小胶质细胞和血清炎性因子白细胞介素1β（IL-1β）、转化生长因子β（TGF-β）、白细胞介素10（IL-10）含量的影响，探讨电针预处理改善缺血性中风后运动功能的机制。方法: C57BL/6小鼠随机分为假手术组、模型组、电针预处理组（电针组），每组15只。光栓诱导构建单侧缺血性卒中运动障碍模型。电针组小鼠在造模前进行连续7 d的电针“百会”“大椎”预处理，每次20 min，每天1次。用网格实验和爬杯实验评价小鼠造模前后运动功能；激光散斑血流监测视频系统评估小鼠初级运动皮层血流灌注量；ELISA法检测小鼠血清IL-1β、TGF-β、IL-10的含量；免疫荧光染色法检测小鼠初级运动皮层小胶质细胞及其M2亚型细胞标记物的表达。结果: 造模后，与假手术组比较，模型组小鼠网格错误率、患肢拖拽率升高（P<0.01），患肢利用率及患侧占健侧皮层血流灌注量百分比降低（P<0.01）；血清IL-1β、TGF-β、IL-10含量升高（P<0.01，P<0.05）；患侧初级运动皮层小胶质细胞呈阿米巴样，离子钙接头蛋白分子1（IBA1）和CD206荧光强度升高（P<0.01）。与模型组比较，电针组小鼠网格错误率、患肢拖拽率降低（P<0.01），患肢利用率及患侧与健侧血流灌注量百分比升高（P<0.05）；血清IL-1β含量降低（P<0.01），TGF-β、IL-10含量升高（P<0.01）；患侧初级运动皮层小胶质细胞胞体较模型组更圆、更密集，IBA1和CD206荧光强度升高（P<0.01）。结论: 电针预处理“百会”“大椎”可提高梗死局部小胶质细胞尤其是M2亚型标记物的表达，提高血清抗炎因子含量及降低促炎因子含量，从而减轻炎性反应。.

UNASSIGNED: The protective effects of electroacupuncture (EA) preconditioning against myocardial ischemia-reperfusion injury (MIRI) have been reported. However, the underlying mechanism remains unclear. Recent research has indicated that the dynamic inflammatory response following MIRI plays an essential role in the progression of myocardial injury. This study aimed to investigate the myocardial protective effects of EA preconditioning on MIRI in rats and to explore the relevant mechanism from the perspective of dynamic inflammatory response.
UNASSIGNED: A MIRI model was employed, and the rats were subjected to EA on Neiguan for four days prior to modeling. The myocardial protective effect of EA preconditioning was evaluated by echocardiography, Evans blue and triphenyltetrazolium chloride staining. Real-time polymerase chain reaction, Western blot, hematoxylin & eosin staining, and immunohistochemistry were utilized to detect the content of mitochondrial DNA, NOD receptor family protein 3 (NLRP3) inflammasome activation, neutrophil recruitment and macrophage infiltration in blood samples and myocardium below the ligation.
UNASSIGNED: We found that EA preconditioning could accelerate the recovery of left ventricle function after MIRI and reduce the myocardial infarction area, thereby protecting the myocardium against MIRI. Furthermore, EA preconditioning was observed to ameliorate mitochondrial impairment, reduce the level of plasma mitochondrial DNA, modulate NLRP3 inflammasome activation, attenuate neutrophil infiltration, and promote the polarization of M1 macrophages towards M2 macrophages in the myocardium after MIRI.
UNASSIGNED: EA preconditioning could reduce plasma mtDNA, suppress overactivation of the NLRP3 inflammasome, facilitate the transition from the acute pro-inflammatory phase to the anti-inflammatory reparative phase after MIRI, and ultimately confer cardioprotective benefits.

OBJECTIVE: To observe the effect of different intensities of electroacupuncture (EA) preconditioning on car-diac function and polarization state of macrophages in mice with acute myocardial ischemia (AMI), so as to explore its possible mechanism underlying improvement of AMI.
METHODS: A total of 50 male C57BL/6J mice were randomly divided into sham ope-ration, AMI model, and EA pretreatment groups (0.5 mA, 1 mA, 3 mA subgroups), with 10 mice in each group/subgroup. The mice in the EA pretreatment groups were subjected to EA stimulation of bilateral \"Neiguan\"(PC6) with 0.5, 1.0 and 3 mA respectively and frequency of 2 Hz/15 Hz for 20 min, once a day, for 3 days. The acute myocardial ischemia model was established by ligating the anterior descending branch (ADB) of the left coronary artery, while the sham operation only had a surgical suture trans-passed below the ADB but without ligation. The myocardial infarction area was measured after TTC staining, and the cardiac function ［left ventricular ejection fraction (EF), short-axis contraction rate (FS)］ was detected by using echocardiography. The M1 macrophages were labeled with CD11b+F480+CD206low, M2 macrophages were labeled with CD11b+F480+CD206high and detected by using flow cytometry, and the expression levels of myocardial interleukin-1β (IL-1β), tumor necrosis factor-α (TNF-α), Toll-like receptor-4 (TLR4) proteins were detected by using Western blot.
RESULTS: Compared with the sham operation group, the model group had a significant increase in the infarction area (P<0.000 1), number of cardiac macrophages and percentage of M1 type macrophages (P<0.000 1), and the expression levels of myocardial IL-1β, TNF-α, TLR4 proteins (P<0.001, P<0.01), and a remarkable decrease in the levels of EF, FS and the percentage of M2 type macrophages (P<0.000 1). In contrast to those of the model group, the area of myocardial infarction (P<0.000 1, P<0.01), expression levels of myocardial IL-1β, TNF-α, TLR4 proteins (P<0.01, P<0.05, P<0.001) in the 0.5 mA, 1 mA and 3 mA groups, number of macrophages and percentage of M1 macrophages (P<0.05) in the 1 mA group were significantly decreased, while the levels of EF and FS (P<0.000 1, P<0.05, P<0.001) in the 3 EA groups, and percentage of M2 macrophage (P<0.05) in the 1 mA group were significantly increased. Comparison among the 3 EA groups displayed that the effects of 1 mA group were significantly superior to those of 0.5 and 3 mA groups in up-regulating EF and FS (P<0.01, P<0.001), and in down-regulating the area of infarct myocardium (P<0.01, P<0.000 1), and the expression of TLR4 protein (P<0.01), and 0.5 mA group in the expression of IL-1β and TNF-α proteins (P<0.05).
CONCLUSIONS: EA preconditioning with electrical current intensities of 0.5 mA, 1 mA and 3 mA can effectively reduce myocardial infarction size, improve cardiac function in mice with AMI, which may be related with its effects in reducing the number of cardiac macrophages and down-regulating the expression of myocardial IL-1β, TNF-α and TLR4 proteins. The therapeutic effect of 1 mA is better than that of 0.5 and 3 mA.
目的：观察不同强度电针预处理对急性心肌缺血（AMI）小鼠心功能影响的效应差异，并探讨其可能的作用机制。方法：C57BL/6J小鼠随机分为假手术组、模型组、电针预处理组(0.5 mA组、1 mA组、3 mA组)，每组10只。各电针预处理组采用0.5、1、3 mA电流强度分别电针小鼠双侧“内关”，每次20 min，每日1次，连续干预3 d后造模。采用结扎心脏冠状动脉左前降支法建立AMI小鼠模型。采用超声心动评价各组小鼠心脏功能，TTC染色法检测心肌梗死面积百分比，流式细胞技术检测心脏巨噬细胞数量和M1/M2巨噬细胞极化状态，Western blot法检测心肌组织白细胞介素-1β(IL-1β)、肿瘤坏死因子α(TNF-α)、Toll样受体4(TLR4)蛋白表达水平。结果：与假手术组比较，模型组左心室射血分数(EF)和短轴收缩率(FS)降低(P<0.000 1)，心肌梗死面积百分比升高(P<0.000 1)，心脏巨噬细胞数量增加(P<0.000 1)，且以M1型为主，心肌组织IL-1β、TNF-α、TLR4蛋白表达水平升高(P<0.001, P<0.01)。与模型组比较，0.5 mA组、1 mA组和3 mA组EF和FS均明显升高(P<0.000 1，P<0.05，P<0.001)，心肌梗死面积百分比明显降低(P<0.000 1，P<0.01)，心肌组织IL-1β、TNF-α、TLR4蛋白表达水平均降低(P<0.01，P<0.05，P<0.001)，1 mA组心脏巨噬细胞数量降低(P<0.05)，且以M1型为主。与1 mA组比较，0.5 mA组和3 mA组EF和FS降低(P<0.01，P<0.000 1)，心肌梗死面积百分比升高(P<0.01，P<0.000 1)，心肌组织TLR4蛋白表达水平升高(P<0.01)，0.5 mA组心肌组织IL-1β、TNF-α蛋白表达水平升高(P<0.05)。结论：0.5、1、3 mA电针预处理均能有效改善AMI小鼠心功能，减少AMI小鼠心肌梗死面积和心脏巨噬细胞数量，下调心肌组织IL-1β、TNF-α、TLR4蛋白表达；其中1 mA组效应最佳，可能与其促进心肌巨噬细胞由M1型向M2型极化有关。.

OBJECTIVE: To explore the protective effect and molecular mechanism of electroacupuncture (EA) preconditioning on renal injury in type 2 diabetic rats.
METHODS: Fifty male Wistar rats were randomly divided into control, model, EA, EA+inhibitor, and inhibitor groups, with 10 rats in each group. Diabetes model was established by high fat and high glucose diet and intraperitoneal injection of streptozotocin (40 mg/kg). EA (2 Hz, 1 mA) preconditioning was applied to \"Guanyuan\" (CV4), \"Zhongwan\" (CV12), bilateral \"Zusanli\" (ST36) and \"Fenglong\" (ST40) for 15 min, once every other day for 8 weeks. Rats of the inhibitor and EA+inhibitor groups were given intraperitoneal injection of 3-TYP (50 mg/kg) once every other day for a total of 3 times. The body weight, kidney mass, and renal index were recorded. The contents of urine microalbumin (ALB), 24 h urine 8-hydroxydeoxyguanosine (8-OHdG) and activities of reactive oxygen species (ROS), superoxide dismutase (SOD), catalase (CAT), glutathione glycine peroxidase (GSH-Px) in kidney were detected by ELISA. The activities of mitochondrial respiratory chain enzyme complex (RCCⅠ-RCCⅣ) in kidney were detected using spectrophotometric method. HE staining, Masson staining and transmission electron microscopy were used to observe the changes of renal structure. The protein and mRNA expressions of silent information regulator 3 (Sirt3), manganese superoxide dismutase (MnSOD) in kidney were detected by Western blot and quantitative real-time PCR, respectively.
RESULTS: After modeling and compared with the control group, the contents of ALB, the renal index, activity of ROS and content of 8-OHdG, and the renal collagen volume fraction (CVF) were increased (P<0.01), while the activities of SOD, CAT and GSH-Px, RCCⅠ-RCCⅣ, and the mRNA and protein expressions of Sirt3 and MnSOD were decreased (P<0.01). After the treatment and compared with the model group, the contents of ALB, the renal index, ROS, 8-OHdG, and the CVF were decreased in the EA group(P<0.01, P<0.05), while the activities of SOD, CAT, GSH-Px, RCCⅠ-RCCⅣ, and Sirt3 and MnSOD expression levels were increased (P<0.01, P<0.05)；the RCCⅡ activity and the expression level of MnSOD mRNA were increased (P<0.05) in the EA+inhibitor group; the ALB and 8-OHdG contents and the CVF in the inhibitor group were increased (P<0.05), while the activity of SOD, and Sirt3 and MnSOD expression levels were decreased (P<0.05). In comparison with the EA group, the contents of ALB, the renal index, activities of ROS and 8-OHdG contents, and the CVF were increased (P<0.05, P<0.01), activities of SOD, CAT and GSH-Px and RCCⅠ and RCCⅡ, and the mRNA and protein expressions of Sirt3 and MnSOD were decreased (P<0.05, P<0.01) in both EA+inhibitor group and inhibitor group, whereas the activities of RCCⅢ and RCCⅣ were decreased in the inhibitor group (P<0.05). The therapeutic effect of inhibitor was notably inferior to that of EA+inhibitor in decreasing ALB and 8-OHdG contents, and CVF (P<0.01), and in up-regulating SOD and RCCⅡ activities, Sirt3 and MnSOD expression levels (P<0.01, P<0.05).
CONCLUSIONS: EA preconditioning can increase the expressions of renal Sirt3 and MnSOD in type 2 diabetic rats, thereby reducing the oxidative stress response and protecting the kidneys.
目的：观察电针预处理对2型糖尿病大鼠肾脏的保护效应和对沉默信息调节因子3(Sirt3)、锰超氧化物歧化酶(MnSOD)表达的影响, 探讨其作用机制。方法：Wistar大鼠随机分为对照组、模型组、电针组、电针+抑制剂组、抑制剂组, 每组10只。除对照组外, 各组大鼠均予高糖高脂饮食联合链脲佐菌素复制2型糖尿病模型。造模前, 电针组、电针+抑制剂组给予电针“足三里”“关元”“丰隆”“中脘”, 15 min/次, 隔日1次, 共8周；电针+抑制剂组、抑制剂组给予腹腔注射Sirt3抑制剂3-TYP(50 mg/kg), 隔日1次, 共3次。测量大鼠双肾质量及体质量, 计算肾脏指数；ELISA法检测尿微量白蛋白(ALB)、24 h尿液中8-羟基脱氧鸟苷(8-OHdG)和肾脏组织活性氧(ROS)、超氧化物歧化酶(SOD)、过氧化氢酶(CAT)、谷胱甘肽过氧化物酶(GSH-Px)活性；紫外分光光度法检测肾脏组织线粒体呼吸链酶复合物(RCCⅠ—RCCⅣ)活性；HE染色法、Masson染色法和透射电镜观察肾脏病理变化；Western blot法和荧光定量PCR法检测肾脏Sirt3、MnSOD蛋白和mRNA表达水平。结果：与对照组比较, 模型组尿ALB含量及肾脏指数、肾脏组织ROS活性、24 h尿液中8-OHdG含量和肾脏胶原容积分数(CVF)升高(P<0.01), 肾脏组织SOD、CAT、GSH-Px活性和RCCⅠ—RCCⅣ活性、Sirt3和MnSOD蛋白和mRNA表达水平降低(P<0.01)。与模型组比较, 电针组尿ALB含量及肾脏指数、肾脏组织ROS活性、24 h尿8-OHdG含量和肾脏CVF降低(P<0.01, P<0.05), 肾脏组织SOD、CAT、GSH-Px活性和RCCⅠ—RCCⅣ活性、Sirt3和MnSOD蛋白和mRNA表达水平升高(P<0.01, P<0.05)；电针+抑制剂组RCCⅡ活性、肾脏组织MnSOD mRNA表达水平均升高(P<0.05)；抑制剂组尿ALB、24 h尿液中8-OHdG含量及CVF升高(P<0.05), 肾脏组织SOD活性、Sirt3和MnSOD蛋白及mRNA表达水平降低(P<0.05)。与电针组比较, 电针+抑制剂组和抑制剂组尿ALB、肾脏指数、肾脏组织ROS、24 h尿液中8-OHdG及CVF升高(P<0.05, P<0.01), 肾脏组织SOD、CAT、GSH-Px活性和RCCⅠ、RCCⅡ活性及Sirt3、MnSOD蛋白和mRNA表达水平降低(P<0.05, P<0.01), 抑制剂组RCCⅢ、RCCⅣ活性降低(P<0.05)。与电针+抑制剂组比较, 抑制剂组尿ALB、24 h尿液中8-OHdG含量及肾脏CVF升高(P<0.01), 肾脏组织SOD活性、RCCⅡ活性和Sirt3、MnSOD蛋白及mRNA表达水平降低(P<0.01, P<0.05)。模型组和抑制剂组肾小球肥大, 肾脏纤维化严重, 足突大量消失, 电针组和电针+抑制剂组肾脏损伤不同程度减轻。结论：电针预处理能减轻糖尿病所导致的肾脏损伤, 其机制可能与促进Sirt3、MnSOD的表达, 改善肾脏氧化应激有关。.

OBJECTIVE: To observe the effect of electroacupuncture (EA) at \"Zusanli\"(ST36) pretreatment on lung functions, inflammatory response, and levels of angiotensin-converting enzyme 2 (ACE2) and angiotensin (1-7) ［Ang (1-7)］ in rats with sepsis-induced acute lung injury (ALI), so as to explore its mechanisms underlying improvement of ALI.
METHODS: Thirty male SD rats were randomly divided into normal, model and EA groups (n=10 in each group). The sepsis-related ALI model was established by intraperitoneal injection of lipopolysaccharide (LPS, 5 mg/kg). Rats of the EA group received EA (4 Hz/20 Hz, 1-3 mA) stimulation at bilateral ST36 for 30 min, once each day, for 7 days before modeling. The lung functions including forced vital capacity (FVC), forced expiratory volume at 0.1 second (FEV0.1) and FEV0.3 were detected using a respiratory function detector for small animals at 3 h after modeling. The bronchoalveolar lavage fluid (BALF) was collected for assaying the contents of Ang (1-7), tumor necrosis factor-α (TNF-α) and interleukin-1 β (IL-1β) using ELISA. The lung wet/dry weight (W/D) ratio, FEV0.1/FVC, and FEV0.3/FVC were calculated. The histopathological changes of lung tissues were displayed by hematoxylin-eosin (H.E.) staining. The expression of ACE2 and mitochondrial assembly receptor (MasR) mRNAs and proteins in the lung tissue was detected by fluorescence quantitative real-time PCR and Western blot, separately.
RESULTS: Following modeling, the levels of FVC, FEV0.1, FEV0.3, ratio of FEV0.1/FVC and FEV0.3/FVC, content of Ang (1-7) in the BALF, and the expression levels of ACE2 and MasR mRNAs and proteins in the lung tissue were significantly decreased (P<0.01), while the level of W/D ratio and TNF-α and IL-1β contents in the BALF significantly increased (P<0.01) in the model group relevant to the normal group. In comparison with the model group, the levels of FVC, FEV0.1, FEV0.3, ratio of FEV0.1/FVC and FEV0.3/FVC, content of Ang (1-7) in the BALF, and expression levels of ACE2 and MasR mRNAs and proteins in the lung tissue were significantly increased (P<0.05, P<0.01), whereas the level of W/D ratio, and TNF-α and IL-1β contents in the BALF were significantly decreased (P<0.05, P<0.01) in the EA group. H.E. staining showed pulmonary interstitial edema and alveolar septum thickening with severe inflammatory cell infiltration in the model group, which was relatively milder in the EA group.
CONCLUSIONS: EA preconditioning at ST36 can improve pulmonary function in sepsis-related ALI rats, which may be related to its effects in inhibiting inflammatory response and up-regulating ACE2 and MasR expression and Ang (1-7) content in the lung tissue.
目的：观察电针“足三里”预处理对脂多糖(LPS)诱导的脓毒症急性肺损伤(ALI)大鼠炎性反应、血管紧张素转化酶2(ACE2)及血管紧张素(1-7)［Ang(1-7)］的影响, 探讨电针对脓毒症ALI的预防作用和可能机制。方法：将30只雄性SD大鼠随机分为正常组、模型组、电针组, 每组10只。采用腹腔注射LPS(5 mg/kg)复制脓毒症ALI大鼠模型。电针组予双侧“足三里”穴电针预处理, 每日1次, 每次30 min, 连续1周后造模。造模3 h后采用小动物呼吸功能检测仪检测大鼠肺功能；ELISA法检测支气管肺泡灌洗液(BALF)中Ang(1-7)、肿瘤坏死因子-α(TNF-α)、白细胞介素-1β(IL-1β)的含量；计算肺湿/干重比(W/D)；HE染色法观察大鼠肺组织病理变化；Western blot法、实时荧光定量PCR法检测肺组织中ACE2、线粒体组装受体(MasR)蛋白和mRNA的表达。结果：与正常组比较, 模型组肺组织可见明显肺间质水肿、肺泡间隔增厚；用力肺活量(FVC)、第0.1秒用力呼气量(FEV0.1)、第0.3秒用力呼气量(FEV0.3)、FEV0.1占FVC之比(FEV0.1/FVC), FEV0.3占FVC之比(FEV0.3/FVC)均显著降低(P<0.01)；BALF内TNF-α、IL-1β含量显著升高(P<0.01), Ang(1-7)含量显著降低(P<0.01)；W/D显著升高(P<0.01)；肺组织内ACE2、MasR蛋白和mRNA表达水平均显著降低(P<0.01)。与模型组比较, 电针组肺间质水肿及肺泡间隔厚度均有改善；FVC、FEV0.1、FEV0.3、FEV0.1/FVC及FEV0.3/FVC均显著升高(P<0.05, P<0.01)；BALF内TNF-α、IL-1β含量显著降低(P<0.01), Ang(1-7)含量显著升高(P<0.01)；W/D显著降低(P<0.05)；肺组织内ACE2、MasR蛋白和mRNA表达水平均显著升高(P<0.05, P<0.01)。结论：电针“足三里”穴预处理对LPS诱导的脓毒症ALI大鼠具有抗炎、降低肺水肿和对肺功能保护的作用, 其机制可能与电针通过上调ACE2和MasR的表达及Ang (1-7)的含量从而抑制炎性反应有关。.

OBJECTIVE: To observe the effect of electroacupuncture(EA) preconditioning on expression of Caspase-1, Gasdermin D(GSDMD) and interleukin-1β(IL-1β) in myocardial tissue of myocardial ischemia reperfusion injury (MIRI) rats in order to explore its underlying mechanisms in resisting MIRI.
METHODS: Forty male rats were randomly divided into 4 groups: normal control (normal), sham operation (sham), MIRI model and EA groups. The MIRI model was established by ligation of the left anterior descending branch of the left coronary artery for 30 min and perfusion. EA (2 Hz/100 Hz, 1 mA) was applied to bilateral \"Neiguan\" (PC6) for 20 min, once a day for 3 consecutive days. The echocardiography was used to analyze the left ventricular end-diastolic dimension (LVEDD), left ventricular end-systolic dimension (LVESD) and left ventricular ejection fraction (LVEF, by using Teichholz formula) 4 h after modeling. The myocardial TTC staining was used to observe the proportion of the infarct area, and Western blot was used to detect the expression levels of GSDMD, Caspase-1, IL-1β proteins in the myocardium.
RESULTS: Compared with the normal group, the immunoactivity of GSDMD was increased in the sham group (P<0.05). Compared with the sham group, the LVEF was significantly decreased (P<0.000 1), while the myocardial infarction area, immunoactivity of GSDMD, and the expression levels of Caspase-1, GSDMD and IL-1β proteins were considerably increased in the model group (P<0.000 1, P<0.001). In comparison with the model group, the decreased ejection fraction and the increased myocardial infarction area, and Caspase-1, GSDMD and IL-1β expression were reversed in the EA group (P<0.001, P<0.000 1, P<0.01).
CONCLUSIONS: EA preconditioning may ameliorate myocardial injury in MIRI rats which may be associated with its function in down-regulating the expression of myocardial Caspase-1 protein to reduce cardiomyocyte pyroptosis.
目的：观察电针预处理对心肌缺血再灌注损伤(MIRI)大鼠心肌细胞焦亡相关蛋白半胱氨酸蛋白酶-1(Caspase-1)、Gasdermin D(GSDMD)、白细胞介素-1β(IL-1β)的影响, 探讨电针预处理减轻MIRI损伤的可能机制。方法：SD大鼠随机分为空白组、假手术组、模型组、电针组, 每组10只。电针组电针大鼠双侧“内关”, 每次20 min, 每日1次, 连续3 d。采用冠状动脉左前降支结扎法复制MIRI大鼠模型。采用超声心动图观察造模4 h后大鼠左心室射血分数(LVEF)以评估心功能；TTC染色法观察大鼠心肌梗死面积；免疫组织化学法观察大鼠心肌组织中GSDMD表达情况；Western blot法检测心肌组织GSDMD、Caspase-1、IL-1β蛋白表达水平。结果：与空白组相比, 假手术组心肌组织GSDMD表达升高(P<0.05)。与假手术组相比, 模型组大鼠术后4 h时LVEF明显下降(P<0.000 1), 梗死面积增大(P<0.000 1), 心肌组织GSDMD表达显著升高(P<0.000 1), 心肌GSDMD、Caspase-1、IL-1β蛋白表达升高(P<0.001, P<0.000 1)。与模型组相比, 电针组大鼠术后4 h时LVEF升高(P<0.001), 梗死面积缩小(P<0.000 1), 心肌组织GSDMD表达降低(P<0.001), 心肌GSDMD、Caspase-1、IL-1β蛋白表达降低(P<0.01, P<0.001)。结论：电针预处理可能通过下调Caspase-1蛋白的表达, 降低心肌细胞焦亡水平, 从而改善MIRI损伤。.

OBJECTIVE: To explore the protective effect and molecular mechanism of electroacupuncture (EA) preconditioning on renal injury induced by streptozotocin (STZ) in hyperglycemic mice.
METHODS: Eighty male C57BL/6 mice were randomly divided into 4 groups: control, model, EA and sham EA groups, n=20 in each group. The hyperglycemic model was established by intraperitoneal injection of 0.1% STZ solution (50 mg/kg) for 5 days. EA preconditioning or sham EA was applied at \"Zusanli\" (ST36) and \"Shenshu\" (BL23), once daily for 7 successive days in the EA or sham EA group. Three days after mode-ling, the blood glucose was measured after fasting for 6 hours. The degree of renal injury was observed by HE staining and PAS staining; the expressions of transient receptor potential cation channel 6 (TRPC6) and Nephrin protein in glomerulus were observed by immunohistochemistry; the expressions of TRPC6 and Nephrin protein in renal cortex were detected by Western blot.
RESULTS: Compared with the control group, the blood glucose level was significantly increased (P<0.01), the glomerular cross-sectional area was apparently increased (P<0.000 1), the glomerular capillaries dilated, the matrix proliferated, the brush edge of renal tubules disappeared, the proportion of abnormal renal tubules increased (P<0.000 1), and the expressions of TRPC6 and Nephrin in glomeruli and renal tissue were significantly increased (P<0.01) in the model group. Following EA treatment and compared with the model and sham EA groups, the blood glucose was significantly down-regulated (P<0.01), the renal tissue injury was apparently alleviate, the cross-sectional area of glomerulus was reduced (P<0.05), the brush edge of renal tubules changes were obviously improved, the proportion of abnormal renal tubules decreased (P<0.000 1), and the expressions of TRPC6 and Nephrin in glomerulus and renal tissue decreased (P<0.05) in the EA group.
CONCLUSIONS: EA preconditioning can alleviate renal injury in hyperglycemic mice, which is closely related to its effects in reducing renal TRPC6 and Nephrin expressions and inhibiting podocyte activation.
目的：探讨电针预处理对链脲佐菌素(STZ)诱导的高血糖小鼠肾损伤的保护效应及分子机制。方法： 将C57BL/6小鼠随机分为对照组、模型组、假电针组和电针组，每组20只。模型组、假电针组和电针组小鼠连续5 d腹腔注射STZ(50 mg·kg-1·d-1)进行造模，电针组在造模前于“足三里”及“肾俞”行电针预处理7 d(2 Hz/15 Hz，梯度电流，30 min/次)。各组小鼠在造模完成3 d后，禁食6 h后测血糖值；HE染色法、PAS染色法观察肾脏组织损伤程度；免疫组织化学法检测肾小球瞬时受体电位阳离子通道蛋白6(TRPC6)及Nephrin蛋白阳性表达；Western blot法检测肾皮质TRPC6及Nephrin蛋白表达。结果：与对照组比较，模型组血糖水平明显升高(P<0.01)，肾小球切面面积明显增大(P＜0.000 1)，肾小球毛细血管明显扩张、基质增生，肾小管刷状缘消失，异常肾小管比例升高(P＜0.000 1)，肾小球及肾组织TRPC6、Nephrin表达明显增加(P＜0.01)。与模型组、假电针组比较，电针组血糖明显降低(P＜0.01)，肾组织损伤不明显，肾小球切面面积缩小(P＜0.05)，肾小管刷状缘消失较少，异常肾小管比例降低(P＜0.000 1)，肾小球及肾组织TRPC6、Nephrin表达降低(P＜0.05)。结论：电针预处理可减轻高血糖小鼠肾脏损伤，其保护效应可能是通过抑制高血糖，减少肾脏TRPC6和Nephrin的表达，抑制足细胞活化实现的。.

BACKGROUND: Electroacupuncture is well known for its advantageous neuroanalgesic and therapeutic effects on myocardial ischemia-reperfusion injury. The purpose of the present research was to verify whether electroacupuncture can alleviate bupivacaine-induced myocardial injury.
METHODS: Specific pathogen-free Wistar rats were used to establish the bupivacaine-induced myocardial injury model. Western blot, PCR, transmission electron microscope and enzyme-linked immunosorbent (ELISA) methods were used to evaluate bupivacaine-induced structure injury and dysfunction of the mitochondria as well as the alleviating effects of lipid emulsion, acupoint injection, and electroacupuncture pre-treatment of the oxidase stress response.
RESULTS: Bupivacaine caused structural damage, degradation, and swelling of mitochondria. Furthermore, it reduced adenosine triphosphate (ATP) synthesis and impaired energy metabolism in the mitochondria. Structural and functional impairment of the mitochondria was alleviated via lipid emulsion injection, acupoint injection, and electroacupuncture pre-treatment. Electroacupuncture pre-treatment of PC6 yielded a greater alleviating effect than others approaches. Following electroacupuncture pre-treatment of PC6 point, the number of mitochondria increased; apoptosis was reduced, enzymatic activity of cytochrome C oxidase (COX) and superoxide dismutase and expression of uncoupling protein 2, voltage-dependent anion channel 1, and Bcl 2 were upregulated and SLC25A6, MDA levels were downregulated. Additionally, our findings indicated that electroacupuncture pre-treatment of PC6 point exerted an effect on the mitochondria via the mitochondrial-transcription-factor-A/nuclear-respiratory-factor-1/proliferator-activated-receptor-gamma-coactivator-1 pathway.
CONCLUSIONS: The present study revealed that electroacupuncture pre-treatment of PC6 could effectively alleviate bupivacaine-induced myocardial mitochondrial damage, thereby providing a theoretical basis for clinical studies and applications of this treatment method.

Myocardial ischemia/reperfusion (I/R) is an important complication of reperfusion therapy for myocardial infarction, and trimetazidine is used successfully for treatment of ischemic cardiomyopathy by regulating mitochondrial function. Moreover, electroacupuncture (EA) preconditioning was demonstrated to be cardioprotective in both in vivo rodent models and in patients undergoing heart valve replacement surgery. However, the mechanisms have not been well elucidated. Mitophagy, mediated by the mTORC1-ULK1-FUNDC1 (mTOR complex 1-unc-51-like autophagy-activating kinase 1-FUN14 domain-containing 1) pathway, can regulate mitochondrial mass and cell survival effectively to restrain the development of myocardial ischemia/reperfusion injury (MIRI). In this study, we hypothesized that EA preconditioning ameliorated MIRI via mitophagy. To test this, rapamycin, an mTOR inhibitor, was used. The results showed that EA preconditioning could reduce the infarct size and risk size, and decrease the ventricular arrhythmia score and serum creatine kinase-myocardial band isoenzyme (CK-MB), lactate dehydrogenase (LDH), and cardiac troponin T (cTnT) in MIRI rats. Moreover, it also attenuated MIRI-induced apoptosis and mitophagy accompanied by elevated mTORC1 level and decreased ULK1 and FUNDC1 levels. However, these effects of EA preconditioning were blocked by rapamycin, which aggravated MIRI, reduced adenosine triphosphate (ATP) production, and antagonized infarct size reduction. In conclusion, our results indicated that EA preconditioning protected the myocardium against I/R injury by inhibiting mitophagy mediated by the mTORC1-ULK1-FUNDC1 pathway.

OBJECTIVE: To explore the protective effect and apoptosis-related mechanism of electroacupuncture (EA) preconditioning in the rats with cerebral ischemia-reperfusion injury.
METHODS: Sixty male SD rats, 3 months old, at SPF grade were randomized into a sham-operation group, an ischemia-reperfusion group and an EA preconditioning group, 20 rats in each one. In the ischemia-reperfusion group and EA preconditioning group, the modified MCAO suture-occlusion method was adopted to exert ischemia for 2 h and reperfusion for 3 h, and thus, the models of focal cerebral ischemia-reperfusion injury were prepared on the right side. In the sham-operation group, the right common carotid artery was separated and no more management was given. In the EA preconditioning group, EA at \"Baihui\" (GV 20), \"Shenshu\" (BL 23) and \"Sanyinjiao\" (SP 6) was provided before modeling, with disperse-dense wave, at 2 Hz/100 Hz, 1 mA in intensity. The stimulation for 15 min was taken as one unit (meaning electric stimulation for 10 min and needle retaining for 5 min without electric stimulation). Such preconditioning was repeated continuously for 4 times, totally for 1 h. The neuroethologic condition was assessed in 3 h of reperfusion in each group. TTC staining method was used to determine the percentage of cerebral infarction zone, TUNEL method was to determine the apoptosis index (AI) in hippocampal neuron and the immunohistochemical method (IHC) was to determine the protein expression of p53 and caspase-3.
RESULTS: Compared with the sham-operation group, the neuroethologic score, the percentage of cerebral infarction zone and neuronal AI were all increased obviously in the ischemia-reperfusion group (all P<0.01). Compared with the ischemia-reperfusion group, the neuroethologic score, the percentage of cerebral infarction zone and neuronal AI were all reduced obviously in the EA preconditioning group (all P<0.01). p53\'s nuclei and caspase-3\'s cytoplasms were stained. The positive cells of both of them were brown-yellow in color. In the sham-operation group, the structure of the right hippocampal CA3 neurons of rats was clear, with few positive cells. In the ischemia-perfusion group, the positive expressions of p53 and caspase-3 in the right hippocampal CA3 were increased obviously (P<0.01). Compared with the ischemia-reperfusion group, the positive expressions of caspase-3 and p53 in the right hippocampal CA3 were significantly reduced in the EA preconditioning group (P<0.01).
CONCLUSIONS: Electroacupuncture preconditioning relieves ischemic injury in brain tissue of rats probably through inhibiting the expressions of p53 and caspase-3 to resisting neuronal apoptosis.