From the perspective of "kidney-heart-meridians"， this paper explores the pathogenesis and treatment strategies for sick sinus syndrome （SSS）. The author proposes that the root cause lies in kidney essence deficiency and failure of source qi generation. The key pathogenic mechanisms include qi sinking with cold generation and constrained heart yang. The chronic progression is attributed to impaired transformation of body fluids and blood， leading to stagnation and obstruction of the meridians. Phlegm retention and blood stasis obstructing the heart meridians are considered major factors in disease aggravation. Accordingly， the core treatment principles are to nourish the kidneys and restore source qi， tonify the heart and regulate the meridians. Supplementing qi， lifting sinking， soothing constraint， and promoting yang are employed to halt disease progression， while warming and enriching body fluids and blood aim to harmonize and unblock the meridians to improve prognosis. Strategies such as resolving phlegm， relieving obstruction， activating blood， and dispersing nodules are applied to prevent deterioration. This provides a reference framework for the clinical management of SSS.

Zishen Huoxue decoction (ZSHX) enhances cardiomyocyte viability following hypoxic stress; however, its upstream therapeutic targets remain unclear. Network pharmacology and RNA sequencing analyses revealed that ZSHX target genes were closely associated with mitophagy and apoptosis in the mitochondrial pathway. In vitro, ZSHX inhibited pathological mitochondrial fission following hypoxic stress, regulated FUN14 domain-containing protein 1 (FUNDC1)-related mitophagy, and increased the levels of mitophagy lysosomes and microtubule-associated protein 1 light chain 3 beta II (LC3II)/translocase of outer mitochondrial membrane 20 (TOM20) expression while inhibiting the over-activated mitochondrial unfolded protein response. Additionally, ZSHX regulated the stability of beta-tubulin through Sirtuin 5 (SIRT5) and could modulate FUNDC1-related synergistic mechanisms of mitophagy and unfolded protein response in the mitochondria (UPR^mt) via the SIRT5 and -β-tubulin axis. This targeting pathway may be crucial for cardiomyocytes to resist hypoxia. Collectively, these findings suggest that ZSHX can protect against cardiomyocyte injury via the SIRT5-β-tubulin axis, which may be associated with the synergistic protective mechanism of SIRT5-β-tubulin axis-related mitophagy and UPR^mt on cardiomyocytes.
Mitophagy/drug effects* ; Tubulin/genetics* ; Animals ; Myocytes, Cardiac/metabolism* ; Drugs, Chinese Herbal/pharmacology* ; Sirtuins/genetics* ; Unfolded Protein Response/drug effects* ; Myocardial Ischemia/genetics* ; Rats ; Humans ; Rats, Sprague-Dawley ; Apoptosis/drug effects* ; Male

Objective To discuss the feasibility of establishing the model of combining myocardial ischemic coronary disease with syndrome of kidney deficiency and blood stasis by surgical method.Methods Non-disease of kidney deficiency and blood stasis syndrome model was built by taking the methods of fright, being placed in a cold environment and injection of hydrocortisone. The model of combining the myocardial ischemic coronary disease with the syndrome of kidney deficiency and blood stasis was established through ligating the left anterior descending branch of artery and injecting hydrocortisone. The rats were divided into kidney deficiency and blood stasis group, combination of disease and syndrome group, and normal group, 5 rats in each group. The temperature, weight, heart rate, breathing rate and whole blood viscosity, casson viscosity of the rats in the two groups before and after modeling were observed. According to TCM clinical diagnosis criteria of kidney deficiency and blood stasis syndrome, TCM syndrome characteristics of the two groups were compared.ResultsCompared with normal groups and before modeling, rat temperature dropped and breathing rate increased in kidney deficiency and blood stasis group, combination of disease and syndrome group (P<0.05). Compared with normal group, rat weight decreased or grew slowly, and whole blood viscosity and casson viscosity increased (P<0.05, P<0.01). There were statistical significant differences in whole blood viscosity and electrocardiogram between the two groups (P<0.05).Conclusion There is no obvious difference between TCM syndrome characteristics of the two groups. They all meet the TCM clinical diagnosis criteria of kidney deficiency and blood stasis syndrome.

ObjectiveTo observe the effects of catalpol, the effective component of Rehmanniae Radix, on atrionector cell apoptosis andβ-tubulin in vitro rabbit injured by simulated ischemia reperfusion;To explore the mechanism of treating sick sinus syndrome.MethodsAtrionector cells were collected from newborn rabbits. Cells were divided into 5 groups:normal group, model group, catalpol high, medium and low dose groups. Anoxia and aglycaemia were established to simulate ischemia. Atrionector cellular damage models were established by recovering the supply of oxygen and sugar. Normal control group and model group were given the same volume of culture medium, while catalpol high, medium and low dose groups were given medicine with relevant concentrations (100, 20, 10μg/mL, respectively). ELISA, FCM, laser scanning confocal microscopy were used to observe apoptosis rate andβ-tubulin of atrionector in each group.Results Apoptosis rate of the model group was obviously higher than the normal group (P<0.01), andβ-tubulin cleavage was obvious. Apoptosis rate in catalpol high, medium and low dose groups were significantly lower than that of the model group (P<0.01);β-tubulin structure were significantly more complete compared with the model group;the fluorescence intensity was significantly higher than that of model group (P<0.01).Conclusion Catalpol can inhibit atrionector cellular apoptosis caused by simulated ischemia reperfusion. Its protective effects on atrionectorβ-tubulin may be the mechanism of the treatment of Rehmanniae Radix for sick sinus syndrome.

Objective To observe the effects of ferulic acid on cell apoptosis andβ-tubulinin vitro of rabbit sinoatrial node injured by hypoxia and low glucose, and to explore its mechanism.Methods Model cells were established on sinoatrial node cells from newborn rabbit by performing deprivation of oxygen and glucose. Then the model cells were divided into 5 groups: a normal control group, a model group, ferulic acid high, medium and low dose group. The normal control group and the model group were treated with equal volume of culture medium, and the three volume ferulic acid groups were treated with ferulic acid of 100, 20, and 10μg/ml respectively. Flow cytometry, laser scanning confocal microscopy were used to observe the sinoatrial node apoptosis rate and cytoskeleton proteinβ-tubulin in each group.Results Apoptosis rate of the model group was obviously higher than the normal group (56.95% ± 11.99%vs. 31.45% ± 6.32%,P<0.01), whileβ-tubulin cleavage was significantly lower than the normal group (5.141 ± 0.218vs. 8.035 ± 0.762,P<0.01). Apoptosis rate of ferulic acid high, medium dose group were significantly lower than the model group (24.85% ± 6.34%, 26.70% ± 9.84%vs. 56.95% ± 11.99%,P<0.01),β-tubulin structure were significantly more complete compared with the model group (7.927 ± 0.357, 5.961 ± 0.351vs. 5.141 ± 0.218,P<0.01).Conclusions Ferulic acid can suppress apoptosis of sinoatrial node cells caused by low glucose and oxygen. Protecting the sinoatrial node cell skeleton protein ofβ-tubulin may be one of the mechanisms.

Objective To observe the effects of Tongyang Huoxue Recipe on cytoskeleton proteinβ-tubulin of ischemia reperfusion injured sinoatrial node cell in rabbits;To discuss its mechanisms in the treatment of sick sinus syndrome. Methods Sinoatrial node cells were obtained from newborn rabbit. Oxygen and glucose were deprived to simulate ischemia and were restored to simulate reperfusion. Cells were divided into 5 groups. Tongyang Huoxue Recipe high-, medium-, low dose groups were given corresponding medicine (final concentrations of 100, 20, 10 μg/mL). The normal group and model group were given equal volume of culture medium. Enzyme mark instrument and laser scanning confocal microscopy were used to observe the sinoatrial node cell activity and cytoskeleton protein β-tubulin of each group. Results Living cells of model group decreased significantly compared with normal group (P<0.01), and cytoskeleton proteinβ-tubulin cleavage significantly. Living cells of Tongyang Huoxue Recipe high, medium and low dose group were significantly higher than those in the model group (P<0.05), and β-tubulin structure remained more complete than the model group. Conclusion Tongyang Huoxue Recipe can inhibit the injury induced by ischemia reperfusion of sinoatrial node cells;the possible mechanism of Tongyang Huoxue Recipe in treating sick sinus syndrome is protecting activity and cytoskeleton proteinβ-tubulin of sinoatrial node cells.

Objective To observe the clinical efficacy of Zishen Tongyang Huoxue Formula combined with western medicine in treating angina pectoris caused by coronary heart disease (CHD).Methods Ninety patients were randomly divided into control group, TCM group and combining group, each group of 30 cases. The control group was given western medicine standardized treatment;the TCM group was treated with Zishen Tongyang Huoxue Formula only;the combining group took Zishen Tongyang Huoxue Formula and western medicine standardized treatment. The treatment course for the three groups lasted for four weeks. The clinical presentations, angina pectoris, ECG and serum biomarkers of patients in the three groups were assessed and evaluated before and after treatment.Results The clinical symptoms of TCM were significantly improved;the duration of angina was shortened;the dosage of nitroglycerin reduced after the treatment (P<0.05). The effects in the combining group was more evident compared with the other two groups (P<0.05), but there was no statistical difference between the other two groups (P>0.05). The total TCM effective rates of three groups were 83.33% (25/30), 83.33% (25/30) and 96.67% (29/30), respectively, among which the combining group was more significant (P<0.05). The total ECG effective rates of three groups were 70.00% (21/30), 76.67% (23/30) and 76.67% (23/30), respectively. Hs-CRP, IL-6 and HCY in the three groups decreased after the treatment (P<0.05). The combining group was more significant than the other two groups (P<0.05), but there was no statistical difference between the other two groups (P>0.05). Blood routine test and hepatorenal function of the three groups were all in normal range before and after treatment (P>0.05).Conclusion Using Zishen Tongyang Huoxue Formula combined with western medicinein to treat angina pectoris is effective, safe, and reliable.

Objective To observe the clinical therapeutic efficacy of prescription of supplementing qi and activating yang on sick sinus syndrome. Methods According to the random number table, 231 patients with sick sinus syndrome were randomly divided into treatment group and control group. One hundred and fifty patients in the treatment group were given the Chinese herbal medicine while the other 81 patients in the control group were given atropine. The therapeutic course was four weeks in each group. The changes of clinical manifestations and electrocardiogram (24 h Holter monitoring ECG) of two groups were compared. Results The total effectiveness of treatment group was 98.67% (148/150), the control group was 41.98% (34/81), there was a significant difference in the clinical curative efficacy (P<0.01). The total effective rate of the treatment group was 87.3%(131/150) and that of the control group was 35.9%(29/81), and the average increases of heart rates in 24 hours were 9/min and 4/min, respectively. There was significant difference between the two groups (P <0.05). Conclusion The prescription of supplementing qi and activating yang was effective in improving the clinical symptoms and heart rates of patients with sick sinus syndrome.

OBJECTIVETo compare the effects of enzyme digestion with differential attachment and enzyme digestion with differential attachment plus 5'-bromodeoxyuridine (5-BrdU) (conventional) method for isolating sino-atrial node cells from newborn rats for patch clamp studies.

METHODSThe sino-atrial node tissue were removed from neonatal Wistar rats, digested and separated into single sino-atrial node cells with the above two methods. The obtained cells were observed for morphology, structure, beat frequency, degree of difficulty for patch clamp experiment and the success rate. The atrial cells isolated with enzyme digestion with differential attachment served as the control cells.

RESULTSThe spindle-shaped cells constituted the greatest proportion of the cultured sino-atrial cells, and the cultured atrial cells showed mostly a triangular morphology. Both of the sino-atrial cells separated with the two methods contained mostly spindle-shaped cells, in which spontaneous action potential was recorded. The spindle-shaped cells isolated with enzyme digestion and differential attachment showed a greater viability with a higher success rate in patch clamp studies.

CONCLUSIONThe sino-atrial node cells can be successfully isolated with enzyme digestion and differential attachment, which requires simple procedures and well preserves the cell viability for better performance in patch clamp experiment.

Animals ; Animals, Newborn ; Cell Culture Techniques ; Cell Separation ; methods ; Cells, Cultured ; Patch-Clamp Techniques ; Rats ; Rats, Wistar ; Sinoatrial Node ; cytology

Objective To study the effect of promoting blood circulation to remove blood stasis by Qiangxin Fumai Granule (QXFMG). Method The rat model with blood stasis was induced by the hyodermic injection of adrenaline and ice water-bathing. The blood sample was collected by the puncture of the abdomen aorta and the effect of QXFMG on hemorheological parameters was observed. Result Comparing with the control groups, the QXFMG groups can depress the whole blood viscosity under low shear rate, the erythrocytic clustering index and the content of fibrinogen obviously. Conclusion QXFMG can improve the hemorheology level of rat with acute blood stasis by depressing the whole blood viscosity under low shear rate, restraining the clustering of erythrocyte and depressing the content of fibrinogen, which is prior to the Compound Danshen piece.