miR-214 protects I/R injury in H9c2 cells. the electroacupuncture pretreatment group showed significant decreased myocardial infarct size, as well as the increased indices of Chelerythrine Chloride the cardiac function, including heart rate, mean arterial pressure, left ventricular systolic pressure and maximal rate for left ventricular pressure rising and declining (dp/dt max). In addition, electroacupuncture pretreatment could inhibit the elevation of LDH and CK activities induced by I/R injury. The quantitative PCR (qPCR) results demonstrated electroacupuncture pretreatment could provide cardioprotection against myocardial I/R injury in rats with miR-214 up-regulation. In the meanwhile,in vitro, electroacupuncture pretreatment protected H9c2 Rabbit polyclonal to DDX5 cells from OGD-induced injury. Transfection of miR-214 mimic showed protective effects on OGD-induced injury to H9c2 cells by reducing apoptosis, decreasing LDH and CK activities, rescuing the OGD-induced Ca2+and down-regulating Chelerythrine Chloride elevated protein levels of NCX1, BIM, CaMKII and CypD. == Conclusions == Our findings firstly demonstrated that electroacupuncture pretreatment promotes the expression of miR-214 in myocardial I/R injury and miR-214 contributes to the protective effect of electroacupuncture on myocardial I/R injury. Keywords:I/R injury, miR-214, Electroacupuncture, Protective effect == 1. Introduction == Cardiovascular disease is the leading cause of death in the developed countries and even affects up to 80 million people in the United States.[1]Myocardial ischemia/ reperfusion (I/R) injury contributes to adverse cardiovascular outcomes after myocardial ischemia, cardiac surgery or circulatory arrest.[2]The molecular mechanisms underlying myocardial I/R injury, however, are complex.[3] Although clinical studies have convincingly demonstrated that myocardial ischemia can be effectively treated via acupuncture at the single Neiguan point,[4],[5]the mechanisms in charge of the cardioprotective effect through acupuncture therapy remains undefined.[6] The recent studies have revealed that microRNAs (miRNAs) play important roles in various aspects of cardiac function involved in the regulation of cardiovascular physiological and pathological processes, including cardiac hypertrophy, cardiac fibrosis, cardiac apoptosis and angiogenesis.[7][13]Moreover, it has been found that miR-214 is a sensitive marker of a variety of cardiac stresses, which is up-regulated in I/R-induced cardiac injury and plays a protective role Chelerythrine Chloride against I/R injury by controlling Ca2+overload.[14]Also, acupuncture-serum could decrease Ca2+content in cultured rat myocardial cells,[15]and even electroacupuncture pretreatment was involved in maintaining cardiomyocyte Ca2+homeostasis.[16],[17]However, the role of miRNAs in the cardioprotection by electroacupuncture pretreatment on myocardial I/R injury remains unknown. Therefore, these encourage studies have stimulated us to investigate whether miR-214 is involved in cardioprotection by electroacupuncture pretreatment. == 2. Methods == == 2.1. Myocardial I/R injury model establishment == All procedures involving animals were performed in accordance with the NIH Guide for the Care and Use of Laboratory Animals (NIH Publication no. 86-23, revised 1986). The present study was approved by the Ethics Committee of Southern Medical University. Male Sprague-Dawley rats weighing 280-320 g were obtained from Shanghai Slac Laboratory Animal Company (Shanghai, China). The rats were anesthetized by intraperitoneal injection of 3.5% chloral hydrate (50 mL/kg; ZhuJiang Hospital of Southern Medicine University). Then, the animals were placed in Chelerythrine Chloride the supine position and intubated, ventilated artificially with a respirator (TKR-200C; Jiangxi Teli Anesthesia & Instrument, Nanchang, Jiangxi, China) with a tidal volume of 20 mL/kg body weight and 50 breaths per minute. Electrocardiographic electrodes were connected to the limbs through small needles inserted subcutaneously. Following the skin incision, the hearts were exposed through a left thoracotomy in the fourth intercostal space. The left anterior descending (LAD) coronary artery was ligated with 60 silk suture using a snare occluder. Ischemia was confirmed by visual observation (cyanosis) and continuous ECG monitoring. Evidence for a successful intervention was a cyanotic left anterior ventricular wall and local wall distension, as well as elevated ST segments and peak T waves on the electrocardiogram. After conclusion of 40 min of occlusion, the coronary artery was reperfused by launching the knot. Pursuing reperfusion for 180 min, the rat hearts had been harvested. Sham medical procedures pets underwent the same techniques without occlusion from the LAD. == 2.2. Program of electroacupuncture == The.
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