Network pharmacology for the identification of phytochemicals in traditional Chinese medicine for COVID-19 that may regulate interleukin-6.

OBJECTIVE: ´Three formulas and three medicines,' namely, Jinhua Qinggan Granule, Lianhua Qingwen Capsule, Xuebijing Injection, Qingfei Paidu Decoction, HuaShi BaiDu Formula, and XuanFei BaiDu Granule, were proven to be effective for coronavirus disease 2019 (COVID-19) treatment. The present study aimed to identify the active chemical constituents of this traditional Chinese medicine (TCM) and investigate their mechanisms through interleukin-6 (IL-6) integrating network pharmacological approaches. METHODS: We collected the compounds from all herbal ingredients of the previously mentioned TCM, but those that could down-regulate IL-6 were screened through the network pharmacology approach. Then, we modeled molecular docking to evaluate the binding affinity between compounds and IL-6. Furthermore, we analyzed the biological processes and pathways of compounds. Finally, we screened out the core genes of compounds through the construction of the protein-protein interaction network and the excavation of gene clusters of compounds. RESULTS: The network pharmacology research showed that TCM could decrease IL-6 using several compounds, such as quercetin, ursolic acid, luteolin, and rutin. Molecular docking results showed that the molecular binding affinity with IL-6 of all compounds except γ-aminobutyric acid was < -5.0 kJ/mol, indicating the potential of numerous active compounds in TCM to directly interact with IL-6, leading to an anti-inflammation effect. Finally, Cytoscape 3.7.2 was used to topologize the biological processes and pathways of compounds, revealing potential mechanisms for COVID-19 treatment. CONCLUSION: These results indicated the positive effect of TCM on the prevention and rehabilitation of COVID-19 in at-risk people. Quercetin, ursolic acid, luteolin, and rutin could inhibit COVID-19 by down-regulating IL-6.

[Effect of Xinling Wan in treatment of stable angina pectoris: a randomized, double-blinded, placebo parallel-controlled, multicenter trial].

To evaluate the effectiveness and safety of Xinling Wan on patients with stable angina pectoris, a randomized, double-blinded, placebo parallel-controlled, multicenter clinical trial was conducted. A total of 232 subjects were enrolled and randomly divided into experiment group and placebo group. The experiment group was treated with Xinling Wan (two pills each time, three times daily) for 4 weeks, and the placebo group was treated with placebo. The effectiveness evaluation showed that Xinling Wan could significantly increase the total duration of treadmill exercise among patients with stable angina pectoris. FAS analysis showed that the difference value of the total exercise duration was between experiment group (72.11±139.32) s and placebo group (31.25±108.32) s. Xinling Wan could remarkably increase the total effective rate of angina pectoris symptom score, and the analysis showed that the total effective rate was 78.95% in experiment group and 42.61% in placebo group. The reduction of nitroglycerin dose was (2.45±2.41) tablets in experiment group and (0.50±2.24) tablets in placebo group on the basis of FAS analysis. The decrease of symptom integral was (4.68±3.49) in experiment group and (3.19±3.31) in placebo group based on FAS analysis. Besides, Xinling Wan could decrease the weekly attack time and the duration of angina pectoris. PPS analysis results were similar to those of FAS analysis. In conclusion, Xinling Wan has an obvious therapeutic effect in treating stable angina pectoris, with a good safety and a low incidence of adverse event and adverse reaction in experiment group.

Association of blood pressure and coronary collateralization in type 2 diabetic and nondiabetic patients with stable angina and chronic total occlusion.

OBJECTIVE: We investigated whether and to what extent blood pressure (BP) affects coronary collateralization in type 2 diabetic and nondiabetic patients with stable angina and chronic total occlusion. METHODS: Brachial BP was measured using an inflatable cuff manometer in 431 diabetic and 287 nondiabetic patients with stable angina and angiographic total occlusion of at least one major coronary artery. They were classified according to the SBP (<100, 100-119, 120-139, 140-159, 160-179, and ≥180 mmHg), DBP (<60, 60-69, 70-79, 80-89, 90-99, and ≥100 mmHg), and pulse (<40, 40-49, 50-59, 60-69, 70-79, and ≥80 mmHg) BP ranges. The degree of coronary collaterals supplying the distal aspect of a total occlusion from the contralateral vessel was graded as poor (Rentrop score of 0 or 1) or good collateralization (Rentrop score of 2 or 3). RESULTS: In diabetic patients, the incidence of poor collateralization was related to the DBP in a U-shaped pattern, with the lowest risk at 80-89 mmHg. In nondiabetic patients, an optimal DBP range was 90-99 mmHg for good collaterals, but no U-shaped relation between DBP and coronary collateralization was observed. After adjusting for the baseline characteristics in the logistic regression models, the increased risk of poor collateralization persisted for low or high DBP ranges in diabetic [odds ratio (OR) 2.02-7.29, P ≤ 0.04] and nondiabetic patients (OR 3.62-5.98, P ≤ 0.02). No such relations were observed between collateral grades and SBP and pulse BP. CONCLUSION: This study demonstrates that 80-89 and 90-99 mmHg are the optimal ranges for DBP in diabetic and nondiabetic patients with stable angina and chronic total occlusion, within which the risk of poor collateralization is low.

Impact of losartan and angiotensin II on the expression of matrix metalloproteinase-9 and tissue inhibitor of metalloproteinase-1 in rat vascular smooth muscle cells.

The present study aimed to investigate the impact of losartan and angiotensin II (AngII) on the expression of matrix metalloproteinase-9 (MMP-9) and tissue inhibitor of metalloproteinase-1 (TIMP-1), secreted by rat vascular smooth muscle cells (VSMCs). Rat VSMCs were isolated and cultured in different concentrations of AngII and losartan for 24 h and western blot analysis and quantitative polymerase chain reaction were performed to observe the subsequent impact on the gene and protein expression of MMP-9 and TIMP-1. AngII was shown to promote the protein and gene expression of MMP-9 in VSMCs in a concentration-dependent manner. No effect was observed on the expression of TIMP-1, therefore, an increase in the MMP-9/TIMP-1 ratio was observed. Losartan was shown to be able to inhibit MMP-9 protein and gene expression in a concentration-dependent manner, whilst promoting an increase in TIMP-1 expression, thus decreasing the ratio of MMP-9/TIMP-1. The combined action of losartan and AngII resulted in the same directional changes in MMP-9 and TIMP-1 expression as observed for losartan alone. The comparison of AngII, losartan and the combinatory effect on the expression of MMP-9 and TIMP-1 in VSMCs indicated that losartan inhibited the effects of AngII, therefore reducing the MMP-9/TIMP-1 ratio, which may contribute to the molecular mechanism of losartan in preventing atherosclerosis. In atherosclerosis, the development of the extracellular matrix of plaque is closely correlated with the evolution of AS. The balance between MMPs and TIMPs is important in maintaining the dynamic equilibrium between the ECM, and the renin-angiotensin-aldosterone system, which is involved in the pathologenesis of AS, and in which AngII has a central role.

ERK5 knock down aggravates detrimental effects of hypothermal stimulation on cardiomyocytes via Bim upregulation.

Mechanism of cold induced myocardial injury remained unclear. Our study investigated the role of ERK5/Bim pathway in hypothermal stimulation-induced apoptosis or damage of cardiomyocytes (CMs). Results showed that in CMs which under hypothermal stimulation, ERK5 siRNA promoted expression of Bim protein. Bim siRNA did not influence ERK5 expression but attenuated production of p-ERK5. ERK5 siRNA induced higher apoptosis rate; intracellular Ca(2+) overload; ROS activity; ΔΨm damage in hypothermia stimulated CMs, when compared with hypothermal stimulation solely treated group, while Bim siRNA effected oppositely and canceled pro-apoptotic effect of ERK5 siRNA. In conclusion, ERK5 knock down releases inhibition to Bim expression, induces aggravated apoptosis in CMs under hypothermal stimulation, which related to higher intracellular Ca(2+) overload, ROS activity, and more severe ΔΨm damage. Results revealed regulative role of ERK5/Bim pathway in hypothermal stimulation-induced injure or apoptosis of cardiomyocytes.

Circulating miR-214 is associated with the severity of coronary artery disease.

OBJECTIVE: To study whether miR-214 is regulated in coronary artery disease (CAD) patients and whether placental growth factor (PLGF) is a possible target for miR-214 in atherosclerosis. METHODS: Circulating miR-214 was measured by quantitative PCR using RNA isolated from 40 patients with CAD, including 12 with stable angina pectoris, 16 with unstable angina pectoris and 12 with acute myocardial infarction, and 15 controls without CAD. Plasma level of PLGF was measured by ELISA. RESULTS: The miR-214 level was significantly lower in CAD patients compared with that in controls (P < 0.01). Compared to controls, patients with unstable angina pectoris (UAP, 38.6±9.1 pg/mL) and acute myocardial infarction (AMI, 46.3±13.4 pg/mL) had significantly higher level of plasma PLGF, but not those with stable angina pectoris (SAP; P = 0.012, UAP vs. Control; P = 0.005, AMI vs. Control). In patients with AMI, the plasma level of miR-214 was positively correlated to that of PLGF. CONCLUSIONS: The results suggest that miR-214 is a beneficial microRNA for CAD patients. Loss of its protection may lead to increased PLGF levels and worsening atherosclerosis. Circulating miR-214 is a promising biomarker for alerting severe CAD.

Effects of mid-myocardial pacing on transmural dispersion of repolarization and arrhythmogenesis.

INTRODUCTION: Epicardial (Epi) activation of the left ventricular (LV) wall increases transmural dispersion of repolarization (TDR), which creates a substrate for the development of ventricular arrhythmia. We hypothesize that pacing from the LV mid-myocardium may decrease the TDR and occurrence of arrhythmias. METHODS AND RESULTS: A transmural electrocardiogram and transmembrane action potentials were simultaneously recorded from Epi, mid-myocardial (M), and endocardial (Endo) layers of the arterially perfused canine LV wedge preparations (n= 8). Transmural dispersion of repolarization varied when the preparations were paced at each layer, respectively (Endo pacing, 35.6 ± 6.6 ms; M pacing, 34.9 ± 7.3 ms; Epi pacing, 72.4 ± 4.9 ms; P< 0.001). A significant difference was noted in TDR between M pacing and Epi pacing (P< 0.001), but not between M pacing and Endo pacing (P= 0.831). This result was reproducible in the presence of ischaemia-reperfusion experiments (n= 8). Transmural dispersion of repolarization was amplified as compared with non-ischaemic experiments and differed when preparations were paced at each layer (Endo pacing, 62.8 ± 13.8 ms; M pacing, 63.3 ± 13.3 ms; Epi pacing, 111.1 ± 17.7 ms; P< 0.001). There was again no significant difference between Endo pacing and M pacing (P= 0.948). However, as pacing was shifted from M to Epi, there was a significant increase in TDR (P< 0.001). Ventricular arrhythmias were induced in two of eight ischaemic preparations during Epi pacing, but did not occur in either M or Endo pacing. CONCLUSION: Mid-myocardial pacing can significantly decrease the TDR and prevent the occurrence of ventricular arrhythmias as compared with Epi pacing.

Vitamin E ameliorates ox-LDL-induced foam cells formation through modulating the activities of oxidative stress-induced NF-κB pathway.

The role of antioxidant supplementation with vitamin E in the prevention of atherosclerosis has been a topic of considerable recent interest. The relevance of vitamin E for macrophage-derived foam cell formation, a hallmark of atherosclerosis, however, has not been unequivocally resolved. Here, we investigated the effect of oxidized LDL (ox-LDL) and vitamin E on lipid accumulation and total cholesterol content in U937 macrophages, reactive oxygen species generation and expression of nuclear factor-κB (NF-κB) signaling pathway. The results showed that the mRNA expression and protein levels of P-selectin were evident in U937 macrophages treated with ox-LDL and vitamin E, which indicating that expression of P-selectin is important in macrophage-derived foam cell formation. Moreover, P-selectin changes in ox-LDL-induced foam cell formation can be mediated by vitamin E through activities of nuclear NF-κB activated by serine phosphorylation of NF-κB inhibitor α, suggesting that activation of NF-κB pathway by macrophages may occur. Taken together, these data suggested that vitamin E can prevent ox-LDL-induced foam cell macrophages formation through modulating the activities of oxidative stress-induced NF-κB pathway.

Catheter ablation of cardiac fat pads attenuates Bezold-Jarisch reflex in dogs.

BACKGROUND: Bezold-Jarisch reflex (BJR) plays an important role in the pathophysiology of several cardiovascular disorders. Radiofrequency catheter ablation (RFCA) of the vagal ganglia in cardiac fat pads (FPs) may attenuate BJR. The purpose of this study was to examine the effects of RFCA of the cardiac FPs on veratridine-induced BJR in dogs. METHODS AND RESULTS: This study was performed in 30 pentobarbital-anesthetized and open-chest dogs: control group received no ablation (n = 15); and ablation group (n = 15) received epicardial ablation of the 3 FPs located near the right pulmonary vein, the inferior vena cava, and the aortic root. The BJR was induced by injection of veratridine (15 µg/kg) into the left ventricle. Before injection of veratridine, there were no significant differences in heart rate (HR), systolic arterial pressure (SAP), diastolic arterial pressure (DAP), mean arterial pressure (MAP), left ventricular systolic pressure (LVSP), left ventricle end-diastolic pressure (LVEDP), left ventricular peak systolic and diastolic velocity (±dp/dt(max)) between these 2 groups (P > 0.05). However, the veratridine-induced decrease of HR in ablation group was significantly lower than that in control group (22.9 ± 8.5 bpm vs 93.3 ± 18.4 bpm, P < 0.01). There were no differences in the reduction of SAP, DAP, MAP, LVSP, LVEDP and dp/dt(max) between both groups (P > 0.05). CONCLUSIONS: RFCA of the cardiac FPs significantly attenuated veratridine-induced cardio-vagal component but not the vasodepressor component of the BJR. This might have therapeutic implications in BJR-related disorders such as cardio-inhibitory vasovagal syncope.

Relation of urotensin II levels to lone atrial fibrillation.

Human urotensin II (U-II) is an 11-amino acid peptide that plays an important role in hypertension and coronary artery disease. However, because there is no information on the role of U-II in the development of lone atrial fibrillation (AF), the goal of this study was to clarify the role of U-II in the development of lone AF. The study enrolled 42 patients (42.1 +/- 4.0 years old) with paroxysmal lone AF and 30 healthy gender- and age-matched control subjects. The following factors were measured in blood collected after an overnight fast: glucose, total cholesterol, low-density lipoprotein cholesterol, triglycerides, high-sensitivity C-reactive protein (hs-CRP), U-II, and vascular cell adhesion molecule 1 (VCAM-1). U-II levels were significantly higher in the lone AF than in the control group (4.09 +/- 1.28 vs 2.85 +/- 0.63 ng/ml, p <0.001). VCAM-1 levels were also higher in the lone AF than in the control group (337 +/- 250 vs 218 +/- 117 ng/ml, p = 0.018). In addition, hs-CRP levels were higher in the lone AF than in the control group (0.88 +/- 0.29 mg/dl vs 0.67 +/- 0.31 mg/dl, p = 0.004). Multivariate logistic regression analysis that included U-II, VCAM-1, hs-CP, and conventional AF risk factors showed that only U-II and hs-CRP were independently associated with lone AF. In conclusion, the results indicate that increased levels of U-II are associated with the development of lone AF. Additional studies will be necessary to determine whether the elevation of U-II is the cause or the result of AF.

[Evaluation on cardio-ankle vascular index derived from ultrasound].

OBJECTIVE: The cardio-ankle vascular index (CAVI) could be obtained by measuring pulse wave velocity (PWV) and blood pressure (BP). This method is associated with various technical drawbacks. We evaluated the accuracy and usefulness of CAVI measured by ultrasound via detecting the aortic and ankle flow directly by ultrasonic probe. METHODS: CAVI was determined in 96 subjects [64 male, mean age (41.2 +/- 8.9) years] who took part in the annual check up program in our department by means of the professional equipment (BP-203RPEII, VP-1000, Japan, CAVIp), the M-mode (CAVIm) and color Doppler flow imaging (CAVId). Measurement reproducibility on was obtained by repeat the measurements in 20 subjects choose randomly from the 96 subjects. Carotid ultrasound (CU) was performed to obtain intima-media thickness (IMT) and beta index in all subjects. RESULTS: CAVI obtained by various methods were similar and comparable (CAVIm 7.74 +/- 1.62, CAVId 7.77 +/- 1.59, CAVIp 8.74 +/- 1.57, all P > 0.05). Inter-group and inter-observer variance was negligible (r1 = 0.98, r2 = 0.97). There were also significant correlations between CAVIm and IMT, CAVIm and beta (r1 = 0.824, r2 = 0.812, all P < 0.01), and between CAVId and IMT, CAVId and beta (r1 = 0.815, r2 = 0.813, all P < 0.01). CONCLUSIONS: CAVI could be correctly measured by ultrasound technique.

[High-concentration palmitic acid inhibits the proliferation of peripheral blood-derived human endothelial progenitor cells in vitro].

OBJECTIVE: To study the effects of palmitic acid (PA) on the proliferation of peripheral blood-derived endothelial progenitor cells (EPCs) in vitro. METHODS: The mononuclear cells (MNCs) were isolated from the peripheral blood by Ficoll density-gradient centrifugation. The isolated EPCs were characterized by Di-LDI uptake and FITC-lectin binding assay using laser confocal microscope, and further identified by detection of CD34, CD133 and VEGFR2 expression using flow cytometry. The cultured EPCs were incubated in the presence of PA at the concentrations of 0, 50, 100, 200, 400 and 800 micromol/L for different durations (0, 12, 24, 36, 48 and 60 h). The cell morphology was observed and cell proliferation determined with CCK-8 assay. RESULTS: Incubation with 400 and 800 micromol/L of PA significantly inhibited the proliferative ability of EPCs as compared with the control group (P < 0.05). PA at 400 micromol/L had the strongest effect on the cell proliferation, and this effect was intensified with the passage of time, reaching the peak at 48 h with the growth inhibition rate of 58.59% (P < 0.05). CONCLUSION: High-concentration PA can significantly inhibit the proliferation of EPCs in vitro.

Vicious cycle composed of gut flora and visceral fat: a novel explanation of the initiation and progression of atherosclerosis.

Microorganisms infection was thought to be associated with atherosclerosis (AS), but the results of trials investigating antibiotic therapy in patients with coronary artery disease were controversial. Recently, a series of studies explored the relationship between gut flora and obesity, which showed that western-style diet (high fat) could induce imbalanced ratio of the Firmicutes versus the Bacteroidetes in the gut of germ-free mice and human beings, and gut flora could promote obesity through several novel pathways, such as inhibition of fasting-induced adipocyte factor. Meanwhile, data from some studies confirm that some link exists between the abdominal obesity and satiety centers (hindbrain and hypothalamus) with neurotransmitters and hormones. All of the above showed some connection between western-style diet, gut flora, visceral fat, and satiety centers, but until now, no study has shown us the exact mechanism about it. It is proposed that the vicious cycle composed of gut flora and visceral fat may initiate and promote AS. Attempts to confirm this hypothesis may lead to new directions in the study of the pathogenesis of AS and the development of novel strategies for the treatment of AS.

Dysfunction of AQP7 in the periadventitial fat: A novel trigger of atherosclerosis.

Vascular adventitial lesion as a new etiological factor of atherosclerosis (AS) has been confirmed by the results of several different animal models, and some evidence supports adventitial inflammation as the main cause, but the exact mechanism is still elusive. The data from some studies confirm that some link exists between the adventitial inflammation (which also might happen in the periadventitial fat) and atherosclerotic lesions. Aquaporin7 (AQP7) as an aquaglyceroporin, which regulates the permeation of glycerol through the cell membrane and located in the adipose tissue, shows some relationship with obesity. The result of the studies about AQP7-knockout mice and different expression of AQP7 in the cutaneous abdominal adipose tissue among people with different body types proved this phenomenon. Meanwhile, some degree of dysfunction of AQP7 has been proved in the obese. Until now, no study has shown us the data on the correlation of the expression of AQP7 in the periadventitial fat with the severity of atherosclerotic lesions. It is proposed that dysfunction of AQP7 in the periadventitial fat may trigger the adventitial inflammation. Attempts to confirm this hypothesis may lead to new directions in the study of the pathogenesis of AS and the development of a novel agent for this disorder.

Double-blind, multicenter, active-controlled, randomized clinical trial to assess the safety and efficacy of orally administered nicorandil in patients with stable angina pectoris in China.

BACKGROUND: The efficacy and safety of nicorandil were evaluated in Chinese patients with stable angina pectoris (AP) in a double-blind, multicenter, active-controlled, randomized clinical trial. METHODS AND RESULTS: After a 2-week washout period, 232 patients with stable AP were randomized to receive either nicorandil (5 mg tid; 115 patients) or isosorbide mononitrate (ISMN: 20 mg bid; 117 patients) for 2 weeks. Exercise capacity, number of weekly anginal attacks, nitroglycerin (NTG) consumption, and safety were evaluated. Nicorandil and ISMN significantly prolonged the time to 1 mm ST-segment depression in an exercise tolerance test. Both drugs improved the total exercise time and the time to onset of chest pain. There was no significant difference between the 2 groups. Nicorandil significantly decreased the number of anginal attacks and NTG consumption. ISMN decreased the number of anginal attacks significantly; however, there was no significance in NTG consumption, and the ratio of anginal attack reduction was at least 50% was significantly higher with nicorandil. Nicorandil was well tolerated and there was no safety profile difference compared with ISMN. Thus, nicorandil may have equivalent or better antianginal effect than ISMN. CONCLUSIONS: Nicorandil is beneficial as treatment for AP.