| BackgroudBradyarrhythmia is caused by sinus node or atrioventricular node low self-regulation, or it can caused by cardiac conduction system block.This includes sinus bradycardia, sinoatrial or atrioventricular block (AVB), sick sinus syndrome (sss) and so on. It usually manifested clinically dizziness, amaurosis, heart palpitations, a transient disturbance of consciousness, syncope (seriously, have a A-Adams syndrome) and other symptoms, even death, and it belongs to Symptoms of Traditional Chinese Medicine "Vertigo " "fainting "heart palpitations" "palpitation "and other areas. Currently modern medicine has not found a satisfactory effect of the drugs to treat these arrhythmias, due to the high cost, and must have certain equipment and professional staff and other reasons; pacemaker treatment of the disease is difficult to spread, transgenic and stem cell transplantation in the treatment of this disease is still in experimental stage. In recent years have more research about Shenfu Injection in treating bradyarrhythmia disease and made a lot of achievements, however, research on its mechanism of action is relatively small.PurposeThrough clinical and experimental study, discussion the clinical efficacy, application security and the possible intrinsic mechanism of Shenfu Injection in treating bradyarrhythmia disease, aslo discussion the role of Shenfu Injection in the induction of bone marrow mesenchymal stem cells transform into myocardial cells, and thus restore the conduction of Atrioventricular node. This can provide the prerequisite and basis for the development of biological pacemakers, at the same time to provide the basis for clinical applicationhe in the future. MethodsClinical study:A prospective, simple random, single blind, parallel group study, from December 2009 to March 2011 period, from Guangzhou University of Traditional Chinese Medicine Department of Cardiology, First Affiliated Hospital to receive patients met the diagnostic criteria in 47 cases Were randomly divided into two group,26 in cases treatment group,21 cases in control group. Treatment group using 20ml Shenfu injection, the control group using 20ml Danshen injection for treatment, both of those treatments were continued for a week. Observed the change of traditional Chinese Syndrome, resting electrocardiogram and laboratory indicators about the two groups patients before and after the treatment,4 days after treatment, two groups were randomly selected 10 patients for esophageal pacing ECG examination, In the process of this examination, two groups of patients were intravenous inject shenfu injection and atropine injection, Before treatment, 10min after treatment were detected indicators of patients with heart rate, sinus node recovery time, corrected sinus node recovery time, sinoatrial conduction time and Venturi-type atrioventricular block point, Observed, compared to its changes before and after treatment.Animal Study:The age of 3 to 4 months, weighing 2-2.5kg of 18 New Zealand white rabbits were divided into 3 groups(n=3), each group have 6 rabbits, Without limiting the male and female. First Shenfu injection were used with the high (Shenfu 5ml) and low dose (Shenfu lml), the control group (saline 3ml) were injected throngh rabbit ear vein for 1 week, And then by rabbit bilateral posterior superior iliac spine pumping the bone marrow, and in vitro bone marrow mesenchymal stem cells (MSCs) were isolated, cultured and amplified. The cells were divided into two subgroups (A group, B group),3 cases in each group,24 hours after cell inoculation, began induced differentiation, when cells were induced to differentiate in vitro, in groups with high and low dose of Shenfu injection, A sub-group were in the regular culture medium with different concentrations of Shenfu (250ul/ml,50ul/ml), added in the 5-F cytidine (5-aza), B sub-groups were in the regular culture medium was added 5-fluoro cytidine (5-aza), but without Shenfu injection; in control group, A sub-group plus 5-FU cell Glucoside (5-aza), B sub-group without any drugs. After 24 hours, the cell medium was changed, according to conventional cell culture continuously cultured cells until 28 days。By cell morphology, cardiomyocyte specific proteins (a actin, troponin-T) to identify the effect of induced differentiation, at the same time after the animal model established for 2 weeks apply the Elisa assay to determine the serum of stem cell factor (SCF) and granulocyte colony stimulating factor (G-CSF) in rabbit blood.ResultClinical study:The Traditional Chinese Medicine syndrome score of Shenfu injection treatment group is less than before treatment, and better than Danshen injection group (P<0.05), TCM total effective rate of Shenfu injection treatment group was 88.5%, significantly better than the control group 57.1% (P<0.05); Clinical symptoms, ECG efficacy, compared with the control group, no significant difference; Esophageal pacing ECG Shenfu injection group in a better trend in the indicators, but no significant difference or no statistically significant; Compared with the atropine injection group, the the difference of the two groups in the sinus node recovery time, corrected sinus node recovery time, sinoatrial conduction time were not statistically significant; The atropine injection group not significantly better than Shenfu injection group, indicating that the Shenfu injection can also improve the indicators described above, but the effect was not significant.Animal Study:After 4 weeks of cell culture, preparation climbing film, by HE staining were observed under light microscope:cells close to the form and extent of myocardial cells, from good to bad in the order:Shen Fu-pin high-dose A group> Shen Fu-pin high-dose B group> A reference group of low-dose needle attached to> participate in group B with low-dose needle> control group A group> control group B group. Troponin T test showed that cultured cells: added both Shenfu injection and 5-aza at the same time for inducing bone marrow mesenchymal stem cells transformed into myocardial cells positive intensity higher than that only induced by 5-aza, and have a certain relationship with the dose, the higher the dose, the higher the positive strength of transformed Into myocardial cells; a-actin analysis showed that, added both Shenfu injection and 5-aza at the same time for inducing bone marrow mesenchymal stem cells transformed into myocardial cells positive intensity higher than that only induced by 5-aza, and have a certain relationship with the dose, the higher the dose, the higher the positive strength of transformed Into myocardial cells, however, somewhat less relevant, this may be related to a-actin-specific non-cardiac related, also may be relevanted to small sample size. Rabbit peripheral blood stem cell factor test:Detected by ELISA, the sample concentrations of high dose Shenfu injection group was higher than the control group(P<0.05), that of high dose Shenfu injection group compared with low dose group and Shenfu injection low dose group compared with control group were no Significance statistical difference (P> 0.05);Rabbit peripheral blood of granulocyte colony stimulating factor test:Detected by ELISA, the sample concentrations of high dose Shenfu injection group was higher than the low dose Shenfu injection and the control group(P<0.05), that of low dose Shenfu injection group compared with control group was no Significance statistical difference (P> 0.05);The results showed that:Shenfu injection have a certain stimulus in the production of rabbit bone marrow hematopoietic growth factors-stem cell factor, Granulocyte colony stimulating factor, and the higher drug concentration, the stronger the effect of its stimulus; Those proved that Shenfu injection have a certain stimulating effect for bone marrow hematopoietic, and associated with with the drug dose, the higher the dose, the stronger the effect of its stimulation.ConclusionUseing Shenfu injection in treating bradyarrhythmia disease can significantly improve patient TCM syndrome; Shenfu injection enable increase the rate of conversion of bone marrow mesenchymal stem cells transfor into myocardial cells Significantly; Shenfu injection have a certain stimulus for bone marrow stem cells; the role of Shenfu injection in bradyarrhythmia and bone marrow mesenchymal stem cells transfor into myocardial cells worth further exploration and research.
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