The study was aimed to assess the effect of Klotho gene and sinoatrial node pacing channel gene (HCN4 and HCN2) for studying sick sinus syndrome, with Klotho gene under the interference of Plasmid-mediated short hairpin RNA. Twenty-five C57BL/6J mice were divided into four groups, i. e, plasmid shRNA 24h group, plasmid shRNA 12h group, sodium chloride 24h group and sodium chloride 12h group. Plasmid shRNA 50microL (1microg/microL) and sodium chloride 50microl were respectively injected according to mice vena caudalis into those in plasmid shRNA group and sodium chloride group. After 12h or 24h respectively, all mice were executed and their sinoatrial node tissues were cut. The mRNA of Klotho, HCN4 and HCN2 gene were detected by RT-PCR. The results of RT-PCR showed that Klotho, HCN4 and HCN2 mRNA levels were lower compared with those in sodium chloride 12h group after 12h interference interval. The results indicated that there might be the a certain relationship between Klotho gene and sinoatrial node pacing channel gene.
Animals ; Glucuronidase ; genetics ; Hyperpolarization-Activated Cyclic Nucleotide-Gated Channels ; genetics ; metabolism ; Male ; Mice ; Mice, Inbred C57BL ; Plasmids ; genetics ; Potassium Channels ; genetics ; metabolism ; RNA Interference ; RNA, Messenger ; genetics ; metabolism ; RNA, Small Interfering ; genetics ; Sinoatrial Node ; metabolism ; physiology ; physiopathology

Recent evidence indicates that the voltage clock (cyclic activation and deactivation of membrane ion channels) and Ca2+ clocks (rhythmic spontaneous sarcoplasmic reticulum Ca2+ release) jointly regulate sinoatrial node (SAN) automaticity. However, the relative importance of the voltage clock and Ca2+ clock for pacemaking was not revealed in sick sinus syndrome. Previously, we mapped the intracellular calcium (Cai) and membrane potentials of the normal intact SAN simultaneously using optical mapping in Langendorff-perfused canine right atrium. We demonstrated that the sinus rate increased and the leading pacemaker shifted to the superior SAN with robust late diastolic Cai elevation (LDCAE) during beta-adrenergic stimulation. We also showed that the LDCAE was caused by spontaneous diastolic sarcoplasmic reticulum (SR) Ca2+ release and was closely related to heart rate changes. In contrast, in pacing induced canine atrial fibrillation and SAN dysfunction models, Ca2+ clock of SAN was unresponsiveness to beta-adrenergic stimulation and caffeine. Ryanodine receptor 2 (RyR2) in SAN was down-regulated. Using the prolonged low dose isoproterenol together with funny current block, we produced a tachybradycardia model. In this model, chronically elevated sympathetic tone results in abnormal pacemaking hierarchy in the right atrium, including suppression of the superior SAN and enhanced pacemaking from ectopic sites. Finally, if the LDCAE was too small to trigger an action potential, then it induced only delayed afterdepolarization (DAD)-like diastolic depolarization (DD). The failure of DAD-like DD to consistently trigger a sinus beat is a novel mechanism of atrial arrhythmogenesis. We conclude that dysfunction of both the Ca2+ clock and the voltage clock are important in sick sinus syndrome.
Animals ; Arrhythmia, Sinus/physiopathology ; Atrial Fibrillation/physiopathology ; Bradycardia/physiopathology ; Calcium/*physiology ; Calcium Channels/*physiology ; Dogs ; Humans ; Sick Sinus Syndrome/physiopathology ; Sinoatrial Node/physiology/*physiopathology

The hyperpolarization-activated cyclic nucleotide-gated (HCN) channels modulate and regulate cardiac rhythm and rate. It has been suggested that, unlike the HCN1 and HCN2 channels, the slower HCN4 channel may not exhibit voltage-dependent hysteresis. We studied the electrophysiological properties of human HCN4 (hHCN4) channels and its modulation by cAMP to determine whether hHCN4 exhibits hysteresis, by using single-cell patch-clamp in HEK293 cells stably transfected with hHCN4. Quantitative real-time RT-PCR was also used to determine levels of expression of HCNs in human cardiac tissue. Voltage-clamp analysis revealed that hHCN4 current (I(h)) activation shifted in the depolarizing direction with more hyperpolarized holding potentials. Triangular ramp and action potential clamp protocols also revealed hHCN4 hysteresis. cAMP enhanced I(h) and shifted activation in the depolarizing direction, thus modifying the intrinsic hHCN4 hysteresis behavior. Quantitative PCR analysis of human sinoatrial node (SAN) tissue showed that HCN4 accounts for 75% of the HCNs in human SAN while HCN1 (21%), HCN2 (3%), and HCN3 (0.7%) constitute the remainder. Our data suggest that HCN4 is the predominant HCN subtype in the human SAN and that I(h) exhibits voltage-dependent hysteresis behavior that can be modified by cAMP. Therefore, hHCN4 hysteresis potentially plays a crucial role in human SAN pacemaking activity.
Biological Clocks ; physiology ; Cyclic AMP ; physiology ; Cyclic Nucleotide-Gated Cation Channels ; physiology ; Electrophysiological Phenomena ; HEK293 Cells ; Humans ; Hyperpolarization-Activated Cyclic Nucleotide-Gated Channels ; Muscle Proteins ; physiology ; Patch-Clamp Techniques ; Potassium Channels ; Sinoatrial Node ; physiology ; Transfection

BACKGROUNDShen song Yang xin (SSYX) is a compound of Chinese medicine with the effect of increasing heart rate (HR). This study aimed to evaluate its electrophysiological properties at heart and cellular levels.

METHODSThe Chinese miniature swines were randomly assigned to two groups, administered with SSYX or placebo for 4 weeks (n = 8 per group). Cardiac electrophysiological study (EPS) was performed before and after drug administration. The guinea pig ventricular myocytes were enzymatically isolated and whole cell voltage-clamp technique was used to evaluate the effect of SSYX on cardiac action potential (AP).

RESULTSSSYX treatment accelerated the HR from (141.8 +/- 36.0) beats per minute to (163.0 +/- 38.0) beats per minute (P = 0.013) without changing the other parameters in surface electrocardiogram. After blockage of the autonomic nervous system with metoprolol and atropin, SSYX had no effect on intrinsic HR (IHR), but decreased corrected sinus node recovery time (CSNRT) and sinus atrium conducting time (SACT). Intra cardiac EPS showed that SSYX significantly decreased the A-H and A-V intervals as well as shortened the atrial (A), atrioventricular node (AVN) and ventricular (V) effective refractory period (ERP). In isolated guinea pig ventricular myocytes, the most obvious effect of SSYX on action potential was a shortening of the action potential duration (APD) without change in shape of action potential. The shortening rates of APD(30), APD(50) and APD(90) were 19.5%, 17.8% and 15.3%, respectively. The resting potential (Em) and the interval between the end of APD(30) and APD(90) did not significantly change.

CONCLUSIONSThe present study demonstrates that SSYX increases the HR and enhances the conducting capacity of the heart in the condition of the intact autonomic nervous system. SSYX homogenously decreases the ERP of the heart and shortens the APD of the myocytes, suggesting its antiarrhythmic effect without proarrhythmia.

Action Potentials ; drug effects ; Animals ; Drugs, Chinese Herbal ; pharmacology ; Female ; Guinea Pigs ; Heart ; drug effects ; physiology ; Heart Rate ; drug effects ; Heart Ventricles ; In Vitro Techniques ; Male ; Myocytes, Cardiac ; drug effects ; physiology ; Sinoatrial Node ; drug effects ; physiology ; Swine ; Swine, Miniature

Instantaneous chaometry was defined on uniformity theory constructed by the present authors. The sample data, sinus heart rate data and arrhythmia heart rate data from MIT-BIH were analyzed with instantaneous chaometry (ICM); the situation not being distinguished with HRV can be differentiated with ICM. The normal sinus rhythm was found to be of three evident characteristics: (1) instant returning to zero, (2) stability-stable characteristic of ICM on the initial position, (3) interval of mean: 2-7, variance: 1.5-5. The third characteristic shows that the variability of ICM is necessary. The studies on arrhythmia database showed that arrhythmia cases exhibited no returning to zero, nonstability of ICM on the initial position, too small mean or standard deviation, respectively. Evidently, the arithmetic of ICM is simple; ICM can be easily applied in clinical and pathologic analyses.
Arrhythmias, Cardiac ; physiopathology ; Electrocardiography ; methods ; Heart Rate ; physiology ; Humans ; Male ; Middle Aged ; Signal Processing, Computer-Assisted ; Sinoatrial Node ; physiology

OBJECTIVETo observe effects of acupuncture at Neiguan (PC 6) on function of sinoatrial node, so as to provide experimental basis for clinical application of Neiguan (PC 6) to treatment of heart diseases.

METHODSFifty cases of heart diseases were randomly divided into 2 groups, a no-blocking group (n = 35) and a blocking group (n = 15). In the no-blocking group, sinoatrial node recovery time (SNRT), sinoatrial conduction time (SACT), sinoatrial node effective refractory period (SNERP) and heart rate (HR) were determined by using esophagus-left cardiac atrium regulating pulsation technique before and after acupuncture at Neiguan (PC 6); and in the blocking group, the vegetative nerve was blocked by intravenous injection of Propanolol and Atropine, and then SNRT, SACT, SNERP and intrinsic heart rate (IHR) were detected before and after acupuncture.

RESULTSIn the no-blocking group there were significant differences in SACT, SNERP and HR (all P < 0.05) and no significant difference in SNRT (P > 0.05) before and after treatment. In the blocking group, there were no significant differences in SNRT, SACT and SNERP and a significant difference in IHR before and after acupuncture (P < 0.05).

CONCLUSIONAcupuncture at Neiguan (PC 6) has a significant effect on function of sinoatrial node, and the mechanism is possibly related with the bidirectional regulative action of acupuncture at Neiguan (PC 6) on the autonomic nerve in the sinoatrial node.

Acupuncture Points ; Acupuncture Therapy ; Adult ; Female ; Heart Rate ; Humans ; Male ; Middle Aged ; Refractory Period, Electrophysiological ; Sinoatrial Node ; physiology

Normal rhythm in a healthy human heart originates from the natural biological pacemaker, the sinoatrial (SA) node which locates in the right atrium. SA node dysfunction or atrial-ventricular (AV) conduction block causes improper heart rate (bradycardia). Such dysfunction, if severe enough, is currently treated by implanting an electronic pacemaker which has been well established technically, but there are some limitations and inadequacies. Recently, progress in developing engineered cardiac biopacemakers with use of genes or cells has been made in experimental animal models. The hyperpolarization-activated cyclic-nucleotide-modulated (HCN) channel (pacemaker channel) modulates cardiac automaticity via the hyperpolarization-activated cation current (I(f)). HCN genes have been delivered to animal myocardium via viral vectors or HCN-transferred cells for recreating biological pacemakers. Approaches with non-HCN genes or transplantation of beating cells are also novel and have been investigated for generating cardiac biopacers. This article summarizes the progresses in research on recreation of cardiac biopacemakers. Genetically engineered biological pacemaker holds great promise to potentially cure severe bradycardia if critical issues, such as their stability and longevity, are properly solved.
Biological Clocks ; physiology ; Bradycardia ; therapy ; Genetic Engineering ; Heart ; Heart Rate ; Heart Ventricles ; Humans ; Ion Channels ; Myocardium ; Pacemaker, Artificial ; Sinoatrial Node

In this paper are proposed four heartbeat models which correspond to two different positions of the ectopic pacemaker and two coupling styles of the sinus and ectopic pacemakers. The models computed are based on the modified Ikeda phase resetting model. Most of the heartbeat modes are periodic; they exhibit "Arnold's tongue" structure in parameter plane of perturbing frequency and strength. When the ectopic pacemaker is located in the ventricle, there are bistable dynamic modes. The abnormal hearbeat rhythms of bigeminy and trigeminy in clinical medicine have been observed when coupling strength is weaker in odd resetting or refractory time is longer in even resetting.
Cardiac Pacing, Artificial ; methods ; Computer Simulation ; Electrocardiography ; Heart Rate ; physiology ; Humans ; Models, Cardiovascular ; Sinoatrial Node ; physiology

OBJECTIVETo detect the expression of voltage-gated Na(+) channel (NaCh) isoforms in rat sinoatrial node and explore their functions.

METHODSExpressions of NaCh isoforms Nav1.1, Nav1.2, Nav1.3, Nav1.5, Nav1.6 and Nav1.7 in the rat sinoatrial node were detected by immunohistochemistry. The functional roles of the NaChs were tested by observing the effect of tetrodotoxin, a specific blocker of NaChs, on the intrinsic heart rate of isolated rat working heart.

RESULTSThe tetrodotoxin- sensitive neuronal isoforms Nav1.1, Nav1.6 and Nav1.7 as well as the tetrodotoxin-resistant cardiac isoform Nav1.5 were present in the rat sinoatrial node, and the neuronal isoforms were more abundant than Nav1.5 (P<0.05). The selective blockade of tetrodotoxin-sensitive isoforms (presumably Nav1.1, Nav1.6 and Nav1.7) by 100 nmol/L tetrodotoxin scarcely affected the intrinsic heart rate (0.5-/+2.9%, P>0.05) while blockade of tetrodotoxin-resistant isoform (presumably Nav1.5) by 2 micromol/L tetrodotoxin resulted in an obvious decline in the intrinsic heart rate (22.1-/+2.1%, P<0.001).

CONCLUSIONSNav1.1, Nav1.5, Nav1.6 and Nav1.7 are all present in rat sinoatrial node. Although neuronal isoforms are more abundant, Nav1.5 seems to contribute more to activity of the sinoatrial node.

Animals ; Heart Rate ; drug effects ; physiology ; Immunohistochemistry ; Ion Channel Gating ; drug effects ; physiology ; Male ; NAV1.1 Voltage-Gated Sodium Channel ; NAV1.5 Voltage-Gated Sodium Channel ; NAV1.6 Voltage-Gated Sodium Channel ; Nerve Tissue Proteins ; biosynthesis ; Protein Isoforms ; biosynthesis ; Rats ; Sinoatrial Node ; drug effects ; metabolism ; physiology ; Sodium Channels ; biosynthesis ; Tetrodotoxin ; pharmacology

OBJECTIVETo precisely visualize cardiac anatomic structures and simultaneously depict electro-mechanical events for the purpose of precise underblood intervention.

METHODSIntracardiac high-resolution tissue Doppler imaging was used to map real time myocardial contractions in response to electrical activation within the anatomic structure of the cardiac conductive system using a canine open-chest model.

RESULTSThe detailed inner anatomic structure of the cardiac conductive system at different sites (i.e., sino-atrial, atrial wall, atrial-ventricular node and ventricular wall) with the inside onset and propagation of myocardial velocity and acceleration induced by electrical activation was clearly visualized and quantitatively evaluated.

CONCLUSIONThe simultaneous single modality visualization of the anatomy, function and electrical events of the cardiac conductive system will foster target pacing and precision ablation.

Animals ; Dogs ; Echocardiography, Doppler ; Electrocardiography ; Heart Conduction System ; diagnostic imaging ; physiology ; Myocardial Contraction ; Sinoatrial Node ; diagnostic imaging ; physiology