Mechanism of Sinitang against Myocardial Ischemia-reperfusion Injury： Prediction Based on Network Pharmacology and Verification by Cellular Experiments / 中国实验方剂学杂志

ObjectiveTo predict the mechanism of Sinitang in treating myocardial ischemia-reperfusion injury （MI/RI） based on network pharmacology and verify the prediction results by cellular experiments. MethodThe traditional Chinese medicine system pharmacology database and analysis platform （TCMSP） was employed for retrieval of the main components and potential targets of Sinitang. Online Mendelian Inheritance in Man （OMIM） and GeneCards were employed to obtain the targets of Sinitang in treating MI/RI. STRING was employed to construct the protein-protein interaction （PPI） network， and DAVID to perform gene ontology （GO） annotation and Kyoto Encyclopedia of Genes and Genomes （KEGG） pathway enrichment. Finally， cellular experiments were carried out to verify the predicted anti-MI/RI mechanism of Sinitang. ResultA total of 105 active ingredients and 234 targets of Sinitang were screened out， among which 116 targets were predicted to be involved in the treatment of MI/RI. The GO annotation gave 587 entries， including 417 biological process entries， 101 cell component entries， and 69 molecular function entries. The KEGG analysis enriched 125 signaling pathways， involving vascular endothelial growth factor （VEGF）， phosphoinositide 3-kinase/protein kinase B （PI3K/Akt）， forkhead box transcription factor O （FoxO）， hypoxia-inducible factor-1 （HIF-1） apoptosis and other signaling pathways. The results of cell viability assay showed that Sinitang increased the survival rate of H9C2 cells damaged by hypoxia/reoxygenation （H/R）. Sinitang decreased the levels of tumor necrosis factor-α （TNF-α）， interleukin-6 （IL-6）， and creatine kinase-MB （CK-MB） in H9C2 cells damaged by H/R. The results of flow cytometry demonstrated that Sinitang decreased the apoptosis rate of H9C2 cells damaged by H/R. Western blot showed that Sinitang down-regulated the expression of Bcl-2 related X protein （Bax） and up-regulated that of B-cell lymphoma-2 （Bcl-2） in H/R-injured H9C2 cells. ConclusionSinitang treats MI/RI in a multi-target and multi-pathway manner， which involves the signaling pathways associated with apoptosis.

Long non-coding RNA-ROR aggravates myocardial ischemia/reperfusion injury

Long non-coding RNAs (lncRNAs) play an important role in the pathogenesis of cardiovascular diseases, especially in myocardial infarction and ischemia/reperfusion (I/R). However, the underlying molecular mechanism remains unclear. In this study, we determined the role and the possible underlying molecular mechanism of lncRNA-ROR in myocardial I/R injury. H9c2 cells and human cardiomyocytes (HCM) were subjected to either hypoxia/reoxygenation (H/R), I/R or normal conditions (normoxia). The expression levels of lncRNA-ROR were detected in serum of myocardial I/R injury patients, H9c2 cells, and HCM by qRT-PCR. Then, levels of lactate dehydrogenase (LDH), malondialdehyde (MDA), superoxide dismutase (SOD), and glutathione peroxidase (GSH-PX) were measured by kits. Cell viability, apoptosis, apoptosis-associated factors, and p38/MAPK pathway were examined by MTT, flow cytometry, and western blot assays. Furthermore, reactive oxygen species (ROS) production was determined by H2DCF-DA and MitoSOX Red probes with flow cytometry. NADPH oxidase activity and NOX2 protein levels were measured by lucigenin chemiluminescence and western blot. Results showed that lncRNA-ROR expression was increased in I/R patients and in H/R treatment of H9c2 cells and HCM. Moreover, lncRNA-ROR significantly promoted H/R-induced myocardial injury via stimulating release of LDH, MDA, SOD, and GSH-PX. Furthermore, lncRNA-ROR decreased cell viability, increased apoptosis, and regulated expression of apoptosis-associated factors. Additionally, lncRNA-ROR increased phosphorylation of p38 and ERK1/2 expression and inhibition of p38/MAPK, and rescued lncRNA-ROR-induced cell injury in H9c2 cells and HCM. ROS production, NADPH oxidase activity, and NOX2 protein levels were promoted by lncRNA-ROR. These data suggested that lncRNA-ROR acted as a therapeutic agent for the treatment of myocardial I/R injury.

Effects of Shenshuaikang Enema Liquid on the Wnt/β-catenin pathway for inhibiting the hypoxia reoxygenation injury in HK-2 cells / 中成药

AIM To investigate the protective effect of serum containing Shenshuaikang Enema Liquid (Rhei Radix et Rhizoma,Salviae miltiorrhizae Radix et Rhizoma,Carthami Flos,Astragali Radix) on HK-2 cells injured by hypoxia/reoxygenation (H/R) and its effect on Wnt/β-catenin pathway.MEHTODS Six New Zealand white rabbits randomly divided into control group,Shenshuaikang high dose group and PBS group (n =2) were treated accordingly for serum collection after 3 days' enema.The HK-2 cells injured by hypoxia/reoxygenation were then administered with serum of rabbits from the control group,PBS group,Shenshuaikang groups (high dose,middle dose and low dose groups due to the differently diluted concentrations) respectively.The H/R damage was determined by ROS fluorescence probe,the cellular damage/apoptosis were analyzed by CFSE/PI method and flow cytometry combined with Annexin V-FITC/PI,and the investigation on effects of drugs on expression of Wnt4 mRNA and β-catenin mRNA were accomplished by fluorescence quantitative PCR.RESULTS The fluorescence intensity of intracellular ROS expression was significantly increased after modeling.CFSE/PI double staining showed that the variant Shenshuaikang dose groups displayed obvious proportional mortality superiority to either the control group or PBS group,and the high dose group achieved the lowest mortality.Annexin V-FITC/PI and flow cytometry showed that,at 12 h,compared with the control group (45.6 ± 2.2)％ and PBS group (41.6 ±0.7) ％,Shenshuaikang groups [low dose group (14.8 ± 0.3) ％,middle dose group (10.3 ± 0.6) ％,high dose group (12.9 ± 0.9)％] obviously inhibited apoptosis/death.Shenshuaikang Enema Liquid medicated serum demonstrated its significant effect on the increase of the Wnt4 mRNA expression and a dual-directional regulation on the expression of β-caterin mRNA by quantitative PCR (P ＜ 0.05).CONCLUSION Inhibition of the apoptosis/death of HK-2 cells with hypoxia/reoxygenation injury due to the serum containing Shenshuaikang Enema Liquid suggests the agent's influence on the activation of Wnt/β-catenin pathway.

Effects and mechanism of TG6 on myocardial ischemia and reperfusion injury / 药学实践杂志

Objective To investigate the protective effects and mechanism of TG 6 on myocardial ischemia/reperfusion injury . Methods the protective effects of TG 6 on myocardial ischemia/reperfusion was investigated by setting up models of ischemia and reperfusion of rats induced by ligating the left coronary anterior descending artery in vivo,isolated rat hearts through an improved Lange-ndorff device, and hypoxia /reoxygenation injury of neonatal rat cardiomyocytes , and the serum CK,LDH,T-SOD, MDA were taken as research markers .Results TG6 significantly reduced the myocardial infarct size , decreased the activity of CK and the content of MDA in serum, reduced the activity of LDH, and increased the activity of T-SOD in vivo;TG6 obviously increased the coronary blood flow after low rate perfusion and reperfusion , decreased the content of MDA and the leakage of CK , LDH in myocardial tissue , elevated the activity of T-SOD in vitro of isolated rat hearts;TG6 had no effects on cells in normal growth condition , raised the viability of cardio-myocytes significantly, and reduced the rate of CK leakage and the content of [Ca2+]i obviously in Na2S2O4 treated cells in vitro of neonatal rat cardiomyocytes .Conclusion TG6 could effectively protect myocardial ischemia/reperfusion injury .

Anti-control of sine function-delay feedback for H-R neuron / 医用生物力学

Objective To study the self-adaptive anti-control of a single H-R neuronal system transformed from periodic dynamics pattern to chaos. Methods Numerical analysis was performed by adding sine function self-delay feedback while coupling intensity and time-delay, respectively, as the control parameters. Results By numerical simulation and analysis, it was found that in a certain range of the combination of coupling intensity and time-delay, the time-interval sequences of the dynamical pattern of a single H-R neuronal system could be controlled from a periodical pattern of 3 spikes onto chaos and other periodical patterns. Conclusions The method of self-adaptive feedback of sine function is effective for the anti-control of H-R neuron, and the coupling intensity and time-delay are both important parameters. The particular self-adaptive dynamics of information identification to neuron is reflected in the control process.

The Effect of Superficial Temporal Artery Compression on Intraocular Microcirculation

To increase retinal blood flow, we attempted to increase blood flow of ophthalmic artery which in the major vascular supply to the eyeball. The authors evaluated changes in blood flow of ophthalmic artery and retinal capillary after compression of superficial temporal artery. In 5 normal healthy subjects, the superficial temporal artery was compressed for 10seconds and the blood flow was measured with color doppler imaging and Heidelberg Retinal Flowmeter(HRF). After compression, the mean volume of ophthalmic artery was increased by 59.3% and the mean change of diastolic velocity was significantly increased by 29.6%. Systolic velocity did not changing significantly. For evaluation of retinal microcirculation, we measured volume, flow, velocity in retina and optic nerve head. The relative ratio in changes of volume, flow, velocity were 87.9%, 91.5%, 92.6%, in retina respectively and 110.1%, 140.7%, 139.5%, respectively in optic nerve head. These significant changes were not statistically(P>0.05). In 5 diabetic patients with damaged autoregulatory mechanism, the relative ratio in changes of volume, flow, velocity were 114.25%, 118.30%, 117.6%, respectively. These changes were not statistically significant(P>0.05). Our results indicate that the increase of blood flow in ophthalmic artery by compressing superficial temporal artery did not increase retinal blood flow.