1.Study on the apoptosis-inducing effect of esculetin on acute myeloid leukemia HL-60 cells via regulating the AKT/SKP2/MTH1 pathway
Weihua SONG ; Fuying CHU ; Wei XIE ; Jinliang CHEN ; Ping ZHAO ; Hong QIU ; Jian TAO ; Xiang CHEN
China Pharmacy 2026;37(1):36-41
OBJECTIVE To investigate the apoptosis-inducing effect of esculetin (Esc) on acute myeloid leukemia (AML) HL-60 cells by regulating the protein kinase B (AKT)/S-phase kinase-associated protein 2 (SKP2)/MutT homolog 1 (MTH1) pathway. METHODS AML HL-60 cells were randomly divided into control group (routine culture), Esc low-concentration group (L-Esc group, 25 μmol/L Esc), Esc medium-concentration group (M-Esc group, 50 μmol/L Esc), Esc high-concentration group (H-Esc group, 100 μmol/L Esc), and high-concentration of Esc+ SC79 (AKT agonist) group (100 μmol/L Esc+5 μmol/L SC79). Cell proliferation in each group was detected by MTT assay and colony formation assay. The level of reactive oxygen species (ROS) in cells was measured by using the CM-H2DCFDA fluorescent probe. Cell apoptosis was analyzed by flow cytometry. Western blot assay was performed to detect the expression levels of apoptosis-related proteins [B-cell lymphoma 2 (Bcl-2), Bcl-2-associated X protein (Bax), cleaved caspase-3], AKT/SKP2/MTH1 pathway-related proteins (p-AKT, AKT, SKP2, MTH1), along with the upstream and downstream proteins of AKT phosphatidylinositol 3-kinase (PI3K), cyclin-dependent kinase inhibitor 1 (P21) and cyclin-dependent kinase inhibitor 1B (P27). RESULTS Compared with control group, the cell viability, colony number, and the phosphorylation levels of AKT and PI3K proteins as well as protein expressions of SKP2, MTH1 and Bcl-2 were significantly decreased (P<0.05), while ROS level, apoptosis rate, and the expression levels of Bax, cleaved caspase-3, P21 and P27 proteins were significantly increased (P<0.05). Moreover, the effects of Esc exhibited concentration-dependence (P<0.05). Compared with H-Esc group, above indexes of high-concentration of Esc+ SC79 group were reversed significantly (P<0.05). CONCLUSIONS Esc may promote massive ROS production and induce activation of apoptosis in HL-60 cells by inhibiting the AKT/SKP2/MTH1 pathway, thus inhibiting the proliferation of HL-60 cells.
2.Increasing toe-out angle during drop-landing can diminish risk of inversion injuries among individuals with chronic ankle instability
Xiaoxue ZHU ; Qiongqiu ZHAO ; Teng ZHANG ; Dan WANG ; Jihong QIU ; Qipeng SONG ; Peixin SHEN
Chinese Journal of Tissue Engineering Research 2025;29(9):1827-1833
BACKGROUND:Individuals with chronic ankle instability are prone to inversion ankle sprains during landing.Moderately increasing the foot toe-out angle during landing may reduce the occurrence of inversion ankle sprains,but no studies have directly demonstrated this effect. OBJECTIVE:To explore the effect of increased toe-out angle during landing on the peak inversion angle,peak angular velocity,and the time to peak inversion among individuals with and without chronic ankle instability. METHODS:A total of 60 participants were recruited for this study,including 30 individuals with chronic ankle instability and 30 without chronic ankle instability.The study utilized a simulated sprain apparatus for drop-landing tests,featuring a platform that could tilt forward by 24° and inward by 15°,thus simulating the foot position during an ankle inversion sprain.Participants were required to perform drop-landing tests under two landing conditions:natural landing and toe-out landing,with the latter involving a greater foot toe-out angle,over 150%more than the former.Kinematic data of participants were recorded using a 12-camera three-dimensional motion capture system.Data analysis was conducted using two-way repeated measures analysis of variance and Spearman correlation analysis. RESULTS AND CONCLUSION:(1)Significant main effects of condition were found for peak inversion angle during drop-landing(P<0.001,η2 p=0.270),peak inversion velocity(P=0.015,η2 p=0.098),and peak inversion time(P<0.001,η2 p=0.260);a significant main effect of group was found for peak inversion velocity(P=0.029,η2 p=0.080).(2)There were significant negative correlations between the foot toe-out angle at landing and the peak ankle inversion angle(P=0.021,r=-0.310;P=0.042,r=-0.278)as well as the peak inversion time(P=0.018,r=-0.312;P=0.021,r=-0.309)in both chronic ankle instability and non-chronic ankle instability groups.Moreover,a significant negative correlation was also found between the foot toe-out angle and peak inversion velocity in the chronic ankle instability group(P=0.021,r=-0.312).(3)It is indicated that increasing the foot toe-out angle at landing can reduce the peak inversion angle,peak inversion velocity,and the peak inversion time during landing in patients with chronic ankle instability and non-chronic ankle instability,thereby decreasing the risk of ankle inversion sprains.
3.Triglyceride-glucose index and homocysteine in association with the risk of stroke in middle-aged and elderly diabetic populations
Xiaolin LIU ; Jin ZHANG ; Zhitao LI ; Xiaonan WANG ; Juzhong KE ; Kang WU ; Hua QIU ; Qingping LIU ; Jiahui SONG ; Jiaojiao GAO ; Yang LIU ; Qian XU ; Yi ZHOU ; Xiaonan RUAN
Shanghai Journal of Preventive Medicine 2025;37(6):515-520
ObjectiveTo investigate the triglyceride-glucose (TyG) index and the level of serum homocysteine (Hcy) in association with the incidence of stroke in type 2 diabetes mellitus (T2DM) patients. MethodsBased on the chronic disease risk factor surveillance cohort in Pudong New Area, Shanghai, excluding those with stroke in baseline survey, T2DM patients who joined the cohort from January 2016 to October 2020 were selected as the research subjects. During the follow-up period, a total of 318 new-onset ischemic stroke patients were selected as the case group, and a total of 318 individuals matched by gender without stroke were selected as the control group. The Cox proportional hazards regression model was used to adjust for confounding factors and explore the serum TyG index and the Hcy biochemical indicator in association with the risk of stroke. ResultsThe Cox proportional hazards regression results showed that after adjusting for confounding factors, the risk of stroke in T2DM patients with 10 μmol·L⁻¹
4.Short- to medium-term safety and efficacy of the implantable Corheart 6 left ventricular assist system in patients with end-stage heart failure
Zhibing QIU ; Xiaochun SONG ; Liangpeng LI ; Hongwei SHI ; Liqiong XIAO ; Yunzhang WU ; Xiaosong RONG ; Jidan FAN ; Liang WEI ; Xin CHEN
Chinese Journal of Clinical Thoracic and Cardiovascular Surgery 2025;32(05):639-645
Objective To investigate the efficacy and safety of the Corheart 6 left ventricular assist system in patients with end-stage heart failure. Methods A retrospective study was conducted on patients with end-stage heart failure who were treated with Corheart 6 left ventricular assist system from March 2022 to June 2024 in 4 hospitals in Jiangsu Province. The efficacy of the device was evaluated by comparing changes in clinical indicators at preoperative, discharge, 3-month postoperative, and 6-month postoperative timepoints, including the New York Heart Association (NYHA) functional classification, left ventricular ejection fraction (LVEF), and left ventricular end-diastolic diameter (LVEDD). The safety of the device was assessed by analyzing the intraoperative position and orientation of the blood pump inlet cannula, as well as the incidence of adverse events. Results In this study, 39 patients were collected, including 34 males and 5 females with a mean age of (56.4±12.5) years, ranging from 20 to 75 years. There was no operative death. There was no death in postoperative 3 months with a survival rate of 100.0%. There were 3 deaths in 6 months postoperatively, with a survival rate of 92.3%. All patients had a preoperative NYHA cardiac function classification of class Ⅳ. The NYHA cardiac function class of the patients improved (P<0.05) at discharge, 3 and 6 months after surgery when compared to the preoperative period. LVEF was significantly higher at 3 months after surgery than that during the preoperative period (P<0.05). LVEDD was significantly smaller at discharge, 3 and 6 months after surgery than that during the preoperative period (P<0.05). The safety evaluation's findings demonstrated that all 39 patients' intraoperative blood pump inlet tubes were oriented correctly, the artificial blood vessel suture sites were appropriate, there were no instances of device malfunction or pump thrombosis, or instances of bleeding or hemolysis, and the rate of the remaining adverse events was low. Conclusion With a low rate of adverse events and an excellent safety profile, the Corheart 6 left ventricular assist system can efficiently enhance cardiac function in patients with end-stage heart failure. It also has considerable clinical uses.
5.Effects of Tongmai Yangxin Pills (通脉养心丸) on Arrhythmia and K+/Ca2+ Channel-Related Proteins and Gene Expression in Myocardial Tissue of Ischemia-Reperfusion Model Rats
Zuoying XING ; Yucai HU ; Huanhuan SONG ; Boyong QIU ; Yankun SONG ; Yongxia WANG
Journal of Traditional Chinese Medicine 2025;66(8):851-859
ObjectiveTo investigate the effects and potential mechanisms of Tongmai Yangxin Pills (通脉养心丸) (TYPs) in preventing ischemia-reperfusion (I/R)-induced arrhythmia. MethodsSixty male SD rats were randomly assigned to sham operation group, model group, amiodarone group, low-dose and high-dose TYPs group, with 12 rats in each group. The sham operation group and the model group received 10 g/(kg·d) normal saline by gavage, the amiodarone group received 60 mg/(kg·d) amiodarone, and the low-dose and high-dose TMP groups received 1 g/(kg·d) and 2 g/(kg·d) TYPs solution respectively, for 21 days, administered twice daily. On the day after the last administration, the I/R model was established in the model and medication groups by ligation of the left anterior descending coronary artery with a cannula, while the sham operation group underwent the same procedure without ligation. Electrocardiogram recordings were continuously monitored throughout the modeling process. Heart rate was recorded at five time points, before ischemia (t-0), 5-10 min after ischemia (t-1), 10-15 min after ischemia (t-2), 15-30 min after ischemia (t-3), and during the first 2 min of reperfusion (t-4); the incidence of arrhythmia including ventricular premature beats (VPB), ventricular tachycardia (VT), and ventricular fibrillation (VF) was recorded; arrhythmia scores were calculated. After 24 hours of reperfusion, left ventricular myocardial tissue was collected. Hematoxylin-eosin (HE) staining was performed to observe pathological changes. RT-PCR was used to detect the mRNA expression of microRNA-1 (miRNA-1), microRNA-133a (miRNA-133a), and potassium (K+) and calcium (Ca2+) ion channel-related genes including KCND2, KCNH2, KCNE2, KCNQ1, KCNE1, KCNJ2, CACNA1C, and CACNB1. Western blot analysis was used to measure protein levels of transient outward potassium current protein (Kv4.2), rapidly activating delayed rectifier potassium current protein (HERG), slowly activating delayed rectifier potassium current protein (KvLQT1), inward rectifier potassium current protein (Kir2.1), and L-type calcium channel protein (Cav1.2). ResultsCompared with sham operation group, the model group showed diffuse myocardial hemorrhage, inflammatory cell infiltration, myocardial necrosis, nuclear pyknosis, vacuolar degeneration, and disrupted myocardial fibers; the model group also exhibited a decreased heart rate (t-1 to t-4), increased arrhythmia scores, elevated miRNA-1 and miRNA-133a expression, and decreased mRNA expression of KCND2, KCNH2, KCNE2, KCNQ1, KCNE1, KCNJ2, CACNA1C, and CACNB1 in myocardial tissue; additionally, Kv4.2, HERG, KvLQT1, Kir2.1, and Cav1.2 protein levels significantly reduced (P<0.01). Compared to the model group, all medication-treated groups showed reduced myocardial damage, including less hemorrhage, reduced inflammatory infiltration, and improved myocardial structure, with the high-dose TYPs group exhibiting the most significant improvement; the amiodarone group and high-dose TYPs group showed a significant increase in heart rate (t-1 to t-4), lower arrhythmia scores, reduced miRNA-1 and miRNA-133a expression; the high-dose TYPs group exhibited significantly increased mRNA expression levels of KCND2, KCNH2, KCNQ1, KCNJ2, and CACNA1C, as well as elevated protein levels of Kv4.2, HERG, KvLQT1, Kir2.1, and Cav1.2 (P<0.05 or P<0.01). ConclusionTMPs can improve myocardial damage and reduce the incidence of ventricular arrhythmia in I/R rats. The underlying mechanism may be related to the downregulation of miRNA-1 and miRNA-133a gene expression, as well as the upregulation of K+ and Ca2+ channel-related genes and proteins.
6.Exploration of Zhuyuwan in Treatment of Atherosclerosis from Perspective of Lipid Transport Disorder
Wei SONG ; Zhongyi ZHANG ; Hairong QIU ; Mei ZHAO ; Zubing ZHOU ; Tao SHEN ; Yong ZHANG
Chinese Journal of Experimental Traditional Medical Formulae 2025;31(13):54-61
This article discusses the key pathogenesis of atherosclerosis (AS) based on the physiological characteristics and pathological changes of lipids and introduces the therapeutic effect of Zhuyuwan on AS, aiming to provide a theoretical basis for the treatment of cardiovascular diseases from the spleen. As essential substances, lipids have the same essence but different forms. They circulate throughout the body with body fluids under the action of Yang Qi to nourish the nutrient Qi and support the defensive Qi. Lipid metabolism disorder often leads to the obstruction of Qi movement, the accumulation of dampness and turbidity, and the generation of phlegm and blood stasis. It has been proven that the formation of vulnerable plaques in AS is attributed to the interaction of three pathogenic factors: deficiency of healthy Qi, phlegm-turbidity, and collateral stasis. Their pathological essence is closely related to abnormal lipid metabolism. As lipids constitute the thick and dense components of body fluids, their impaired dispersion may lead to phlegm-turbidity and blood stasis, the pathological process of which is predominantly ascribed to the dysfunction of the spleen in distributing essence. Therefore, AS is rooted in spleen-stomach disorder, manifests as plaques formed by pathological product accumulation in vessels, with lipid transport disorder as its core pathogenesis. Specifically speaking, the dysfunction of spleen in transportation with accumulation of dampness-turbidity marks the initial stage, and blood turbidity and coagulation and phlegm-nodules accumulating in vessels represent the intermediate phase. Cold accumulation and stagnated heat transforming into toxins represent the terminal stage. Zhuyuwan, first recorded in Taiping Holy Prescriptions for Universal Relief, contains equal proportions of Coptidis Rhizoma and Evodiae Fructus. Coptidis Rhizoma, bitter and cold, exerts descending and purging actions to assist stomach Qi in lowering turbidity. Evodiae Fructus, pungent-bitter and hot, disperses obstruction and promotes free flow to support spleen Qi in ascending the clear. The compatibility of Coptidis Rhizoma and Evodiae Fructus ascends the clear and descends the turbid to harmonize Yin and Yang, assisting the spleen in distributing essence and resolving lipid accumulation to reduce lipid levels. In terms of the therapeutic mechanism, Zhuyuwan modulates lipid metabolism by correcting immune-inflammation network imbalance, improving gut microbiota composition and metabolism, and enhancing reverse cholesterol transport. By analyzing the pathological characteristics of lipid transport disorder in AS, this study delves into the intrinsic connections between cardiovascular disease and lipid transport disorder, giving novel insights into the prevention and treatment of AS.
7.Exploration of Zhuyuwan in Treatment of Atherosclerosis from Perspective of Lipid Transport Disorder
Wei SONG ; Zhongyi ZHANG ; Hairong QIU ; Mei ZHAO ; Zubing ZHOU ; Tao SHEN ; Yong ZHANG
Chinese Journal of Experimental Traditional Medical Formulae 2025;31(13):54-61
This article discusses the key pathogenesis of atherosclerosis (AS) based on the physiological characteristics and pathological changes of lipids and introduces the therapeutic effect of Zhuyuwan on AS, aiming to provide a theoretical basis for the treatment of cardiovascular diseases from the spleen. As essential substances, lipids have the same essence but different forms. They circulate throughout the body with body fluids under the action of Yang Qi to nourish the nutrient Qi and support the defensive Qi. Lipid metabolism disorder often leads to the obstruction of Qi movement, the accumulation of dampness and turbidity, and the generation of phlegm and blood stasis. It has been proven that the formation of vulnerable plaques in AS is attributed to the interaction of three pathogenic factors: deficiency of healthy Qi, phlegm-turbidity, and collateral stasis. Their pathological essence is closely related to abnormal lipid metabolism. As lipids constitute the thick and dense components of body fluids, their impaired dispersion may lead to phlegm-turbidity and blood stasis, the pathological process of which is predominantly ascribed to the dysfunction of the spleen in distributing essence. Therefore, AS is rooted in spleen-stomach disorder, manifests as plaques formed by pathological product accumulation in vessels, with lipid transport disorder as its core pathogenesis. Specifically speaking, the dysfunction of spleen in transportation with accumulation of dampness-turbidity marks the initial stage, and blood turbidity and coagulation and phlegm-nodules accumulating in vessels represent the intermediate phase. Cold accumulation and stagnated heat transforming into toxins represent the terminal stage. Zhuyuwan, first recorded in Taiping Holy Prescriptions for Universal Relief, contains equal proportions of Coptidis Rhizoma and Evodiae Fructus. Coptidis Rhizoma, bitter and cold, exerts descending and purging actions to assist stomach Qi in lowering turbidity. Evodiae Fructus, pungent-bitter and hot, disperses obstruction and promotes free flow to support spleen Qi in ascending the clear. The compatibility of Coptidis Rhizoma and Evodiae Fructus ascends the clear and descends the turbid to harmonize Yin and Yang, assisting the spleen in distributing essence and resolving lipid accumulation to reduce lipid levels. In terms of the therapeutic mechanism, Zhuyuwan modulates lipid metabolism by correcting immune-inflammation network imbalance, improving gut microbiota composition and metabolism, and enhancing reverse cholesterol transport. By analyzing the pathological characteristics of lipid transport disorder in AS, this study delves into the intrinsic connections between cardiovascular disease and lipid transport disorder, giving novel insights into the prevention and treatment of AS.
8.Impact of milk and egg supplementation on body composition and bone mineral density of rural primary school students in Yunnan Province
Chinese Journal of School Health 2025;46(10):1401-1405
Objective:
To investigate the impact of milk and egg supplementation on body composition and bone mineral density of rural primary school students in Yunnan Province, so as to provide a reference for developing targeted nutritional intervention strategies.
Methods:
In December 2023, a cluster sampling method was adopted to select students from grades one to three in four primary schools each from Jinggu and Shidian countys of Yunnan Province, as the intervention group (662 students). Additionally, two boarding primary schools were selected from each county based on the principle of matching scale and student numbers as the control group (455 students). Starting from April 2023, the intervention group received 200 mL milk and 50 g eggs during the break on school days for 8 months, while the control group maintained their usual diet behavior. Body composition was measured by using bioelectrical impedance analysis, and distal radial bone mineral density was assessed via dual energy X-ray absorptiometry in April and December 2023. The intervention effects were analyzed by using a difference in-differences approach.
Results:
The final measurements of body fat percentage, skeletal muscle mass and fat free mass of the intervention group and the control group of primary school students were significantly higher than the baseline values, and the net effect of milk and egg intervention on these body composition indicators was not statistically significant ( P >0.05, both before and after adjustment). In contrast, bone mineral density increased significantly by 0.02 g/cm 2 in the intervention group. The net intervention effect on bone mineral density was statistically significant ( β=0.02, 95%CI =0.00-0.04), and remained significant after model adjustment ( β=0.02, 95%CI =0.00-0.04) (both P < 0.05). Subgroup analysis showed statistically significant effects of the intervention among girls ( β=0.02, 95%CI =0.00-0.04), day students ( β=0.04, 95%CI =0.01-0.07), and students with normal nutritional status ( β=0.02, 95%CI =0.00-0.04) (all P <0.05). No significant effect of milk and egg supplementation was observed on body composition indicators (all P <0.05).
Conclusions
Milk and egg supplementation can improve bone mineral density among rural primary school students in Yunnan Province. It is recommended that rural school aged children should increase intake of milk and eggs to support growth and development.
9.Deoxynivalenol contamination in cereals and bakery products in Shanghai and dietary exposure assessment in pregnant women
Kailin LI ; Baozhang LUO ; Renjie QI ; Hua CAI ; Xia SONG ; Jingjin YANG ; Danping QIU ; Zhenni ZHU ; Yi HE ; Hong LIU
Journal of Environmental and Occupational Medicine 2025;42(10):1170-1176
Background Deoxynivalenol (DON), a priority contaminant for food safety risk monitoring, is produced by Fusarium spp. infesting crops, and its common derivatives are 3-acetyl-DON (3A-DON) and 15-acetyl-DON (15A-DON), which have been shown to possess gastrointestinal toxicity, immunotoxicity, reproductive toxicity, and cytotoxicity. Due to the stable physicochemical properties of the DON family of toxins (DONs), they cannot be effectively removed during food processing, thus following the food chain, entering the human body, and posing health risks. Objective To understand the contamination status of DONs in commercial foods (cereals and bakery products) in Shanghai in 2022–2023, and to assess the exposure risk of DONs in pregnant women by combining their dietary consumption data. Methods Liquid chromatography tandem mass spectrometry (LC-MS/MS) was used to determine the contamination level of DONs in 1 100 food samples (cereals and baked goods) collected in 2022 and 944 samples collected in 2023 from Shanghai. The dietary monitoring data of pregnant women in Shanghai from 2016 to 2017 were adopted. The monitoring employed the food frequency questionnaire distributed among pregnant women through a combination of online telephone enquiry and offline on-site face-to-face survey to estimate their food consumption levels. An exposure assessment model was established to calculate the exposure level to DONs, and the probability distribution of the DONs exposure level in the pregnant women group in Shanghai was obtained by applying @Risk 7.5 software and simulating the calculation according to the Monte Carlo principle. With reference to the tolerable daily intake (TDI) of DONs [1.00 µg·(kg·d)−1] proposed by the Joint FAO/WHO Expert Committee on Food Additives, the risk of exposure to DONs from commercial cereals and bakery products in pregnant women in Shanghai was assessed. Results DONs were detected in cereal and bakery samples collected in 2022 and 2023 with different levels of contamination. The level of DONs in cereal foods in 2023 (mean: 36.33 µg·kg−1) decreased compared to 2022 (mean: 23.64 µg·kg−1). However, the positive rate (71.67%) and level (mean: 51.22 µg·kg−1) of DONs in bakery products increased significantly compared with 2022 (positive rate: 10.00%, mean: 24.39 µg·kg−1). The mean consumption of cereals in 783 pregnant women was 222.48 g·d−1 and the mean consumption of bakery products was 36.07 g·d−1, and there was no statistically significant difference in the intake of all types of cereals and bakery products across the early, middle, and late stages of pregnancy. The modelled intakes of DONs via commercial cereals and bakery products for pregnant women in Shanghai were calculated to be 0.20 and 0.57 µg·(kg·d)−1 in 2022 for the mean level and the 95th percentile level, respectively, and 0.16 µg·(kg·d)−1 and 0.35 µg·(kg·d)−1 in 2023, respectively. The results of the health risk assessment showed that pregnant women in Shanghai had 2.6% and 1.4% probability of exposure to DONs from cereal consumption in 2022 and 2023, respectively. Conclusion The risk of exposure of pregnant women in Shanghai to DONs via commercial cereals and bakery products is relatively low (1.4%-2.6%). However, considering the physical sensitivity of pregnant women, they should avoid consuming moldy grains and appropriately reduce intake of bakery products.
10.Regulatory effect of electroacupuncture at "Neiguan" (PC6) on mitochondrial autophagy during the ischemia and reperfusion phases in rats with myocardial ischemia-reperfusion injury.
Qirui YANG ; Xinghua QIU ; Xingye DAI ; Daonan LIU ; Baichuan ZHAO ; Wenyi JIANG ; Yanhua SONG ; Tong PU ; Kai CHENG
Chinese Acupuncture & Moxibustion 2025;45(5):646-656
OBJECTIVE:
To investigate the regulatory effect of electroacupuncture (EA) at "Neiguan" (PC6) on mitochondrial autophagy in rats with myocardial ischemia-reperfusion injury (MIRI) at different phases (ischemia and reperfusion phases), and to explore the bidirectional regulatory effects of EA at "Neiguan" (PC6) and its potential mechanism.
METHODS:
Forty-five male SD rats were randomly divided into 6 groups according to the random number table method, namely, sham-operation group (n=9), model-A group (n=6), model-B group (n=9), EA-A1 group (n=6), EA-B1 group (n=6), and EA-B2 group (n=9). Except the rats in the sham-operation group, the MIRI model was established in the other groups with the physical ligation and tube pushing method. In the model-A group, the samples were collected directly after ligation, and in the model-B group, the samples were collected after ligation and reperfusion. In the EA-A1 group, EA was delivered while the ligation was performed, and afterwards, the samples were collected. In the EA-B1 group, while the ligation was performed, EA was operated at the same time, and after reperfusion, the samples were collected. In the EA-B2 group, during ligation and the opening of the left anterior descending branch of the coronary artery, EA was delivered, and after reperfusion, the samples were collected. EA was performed at bilateral "Neiguan" (PC6), with a disperse-dense wave, a frequency of 2 Hz/100 Hz, a current of 1 mA, and a duration of 30 min. HE staining was employed to observe the morphology of cardiomyocytes, TUNEL was adopted to detect the apoptosis of cardiomyocytes, transcriptome sequencing was to detect the differentially expressed genes in the left ventricle, JC-1 flow cytometry was to detect the mitochondrial membrane potential (MMP) of cardiomyocytes, Western blot was to detect the protein expression of phosphatase and tensin homolog-induced kinase 1 (Pink1), Parkin and p62 in the left ventricle of rats, and ELISA was to detect the levels of serum creatine kinase isoenzyme (CK-MB) and cardiac troponin I (cTn-I) in the rats.
RESULTS:
Compared with the sham-operation group, the cardiomyocytes of rats in the model-B group were severely damaged, with disordered arrangement, unclear boundaries, broken muscle fibers, edema and loose distribution; and the cardiomyocytes in the EA-B2 group were slightly damaged, the cell structure was partially unclear, the cells were arranged more regularly, and the intact cardiomyocytes were visible. Compared with the sham-operation group, the apoptosis of cardiomyocytes increased in the model-B group (P<0.001); and when compared with the model-B group, the apoptosis alleviated in the EA-B2 group (P<0.001). The differentially expressed genes among the EA-B2 group, the sham-operation group and the model-B group were closely related to cell autophagy and mitochondrial autophagy. Compared with the sham-operation group, MMP of cardiomyocytes was reduced (P<0.001), the protein expression of Pink1, Parkin, and p62 of the left ventricle and the levels of serum CK-MB and cTn-I were elevated in the model B group (P<0.001). In comparison with model-A group, the MMP of cardiomyocytes and the levels of serum CK-MB and cTn-I were reduced (P<0.001, P<0.05), and the protein expression of Pink1 in the left ventricle rose in the EA-A1 group (P<0.01). Compared with the model-B group, MMP of cardiomyocytes increased (P<0.001), the protein expression of Pink1, Parkin, and p62 of the left ventricle, and the levels of serum CK-MB and cTn-I decreased (P<0.001) in the EA-B1 group and the EA-B2 group. When compared with the EA-A1 group, MMP of cardiomyocytes increased (P<0.001), and the protein expression of Pink1, Parkin, and p62 of the left ventricle, and the levels of serum CK-MB and cTn-I decreased in the EA-B1 group (P<0.01).
CONCLUSION
EA at "Neiguan" (PC6) can ameliorate MIRI in rats, which may be achieved through the Pink1/Parkin-mediated mitochondrial autophagy pathway. EA can alleviate myocardial injury by enhancing mitochondrial autophagy at the ischemia phase, and it can reduce reperfusion injury by weakening mitochondrial autophagy at the reperfusion phase.
Animals
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Electroacupuncture
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Male
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Myocardial Reperfusion Injury/metabolism*
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Rats, Sprague-Dawley
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Rats
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Acupuncture Points
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Autophagy
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Humans
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Mitochondria/genetics*


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