Aim To investigate the immunomodulatory effects of sea cucumber fucoidan ( SC-FUC) on macro-phage and the signaling pathways. Methods Cell via-bilities in response to different concentrations of SC-FUC were analyzed by MTT, phagocytosis ability was detected by neutral red,and nitric oxide ( NO) produc-tion was examined by Griess reaction kit. The mRNA expression levels of IL-6 , IL-10 , Toll-like receptors (TLRs) and related signal molecules MyD88, TRIF, NF-κB were assayed by real-time PCR. All the experi-ments were based on murine RAW264. 7 cell line. Re-sults SC-FUC could promote RAW264 . 7 cell prolif-eration, phagocytosis as evidenced by uptake of neutral red and release of NO. The effects were significant at the early stage (6 h and 12 h) . SC-FUC could up-reg-ulate the expression of IL-6 , IL-10 , TLR4 , TLR5 , TLR9. Moreover, mRNA expressions of TLRs signaling molecules were increased, as well as MyD88, TRIF, NF-κB. Conclusions SC-FUC could activate macro-phage, and then promote the immune function by pro-moting production or expression of NO, IL-6, IL-10. It is speculated to be relevant to activated cell surface re-ceptors in macrophage, including TLR4, TLR5, TLR9, and NF-κB signaling pathways.

Objective:To explore the possible anti-atherosclerotic mechanisms of glucose co-transporter-2 inhibitor canagliflozin.Methods:ApoE -/-mice fed on Western diet were randomly assigned into the model group ( n=10) and the canagliflozin group ( n=10). C57BL/6J mice fed on normal diet were chosen as the control group ( n=10). Mice in the canagliflozin group were gavaged with canagliflozin for 14 weeks. The presence and severity of atherosclerosis were evaluated with HE and oil red O stainings in aortic root section slices. PCR assay was performed to determine the mRNA expression levels of nitric oxide synthase. Hepatic transcriptome analysis and hepatic amino acid detection were conducted using RNA-seq and targeted LC-MS, respectively. Results:HE staining and oil red O staining of the aortic root showed that AS models were successfully established in ApoE -/-mice fed on Western diet for 14 weeks. Canagliflozin alleviated the severity of atherosclerosis in pathology. Hepatic transcriptome analysis indicated that canagliflozin impacted on amino acid metabolism, especially arginine synthesis in ApoE -/-mice. Targeted metabolomics analysis of amino acids showed that canagliflozin reduced hepatic levels of L-serine, L-aspartic acid, tyrosine, L-hydroxyproline, and L-citrulline, but raised the hepatic level of L-arginine. Compared to the model group, the canagliflozin group exhibited higher serum arginine and nitric oxide levels as well as elevated nitric oxide mRNA expression in aortic tissues ( P<0.05). Conclusion:Canagliflozin regulated the amino acid metabolism, reduced the levels of glucogenic amino acids,and promoted the synthesis of arginine in atherosclerotic mice.

Objective:To explore the effects of canagliflozin on cardiac function and its regulation of ferroptosis in rats with heart failure with preserved ejection fraction (HFpEF).Methods:Thirty-two 7-week-old Dahl salt-sensitive rats were selected and randomly divided into four groups: the control group (fed with low-salt diet), the HFpEF group (fed with high-salt diet), the canagliflozin 20 group (fed with high-salt diet and 20 mg·kg -1·d -1 canagliflozin), and the canagliflozin 30 group (fed with high-salt diet and 30 mg·kg -1·day -1 canagliflozin). Body weight and blood pressure of the rats in each group were monitored. Metabolic cage tests were conducted at the10 th week of the experiment, and echocardiography was performed at the 12 th week, after which the rats were killed. Blood and left ventricular samples were collected. HE staining, Masson staining, Prussian blue iron staining, and reactive oxygen species staining were performed to observe the cardiomyocyte size and shape, degree of interstitial fibrosis, iron staining, reactive oxygen species production under optical microscope. The ultrastructure of cardiomyocytes was observed under electron microscope. Western blotting and real-time fluorescent quantitative reverse transcription polymerase chain reaction (RT-qPCR) were used to detect the expression levels of proteins and mRNA related to ferroptosis in left ventricular myocardial tissue of rats in each group. Results:After 1 week of adaptive feeding, all rats survived. Metabolic cage results showed that compared with control group, rats in the HFpEF group, canagliflozin 20 group and canagliflozin 30 group had more food intake, water intake and urine output, and lower body weight (all P<0.05). These changes were more pronounced in canagliflozin 20 group and canagliflozin 30 group than in HFPEF group, and only the body weight at the 12 th week showed a statistically significant difference between canagliflozin 20 group and canagliflozin 30 group ( P<0.05). The blood pressure of 6 th week and 12 th week, heart weight and left ventricular corrected mass of 12 th week of rats in HFpEF group were higher than those in control group, canagliflozin 20 group and canagliflozin 30 group, while the ratio of early mitral valve peak velocity to late mitral valve peak velocity of 12 th week was lower (all P<0.05). HE and Masson staining showed that compared to control group, the myocardial fibers in the left ventricular myocardial tissue of rats in HFpEF group were disordered, with larger cell diameter ((0.032±0.004) mm vs. (0.023±0.003) mm, P<0.05), irregular shape, obvious proliferation of interstitial collagen fibers, and higher collagen volume fraction (0.168±0.028 vs. 0.118±0.013, P<0.05). Compared with HFpEF group, rats in the canagliflozin 20 group and canagliflozin 30 had more orderly arranged myocardial fibers, more regular cardiomyocyte shape, smaller cell diameter, and lower collagen volume fraction ( P<0.05). It was observed under electron microscopy that, compared to control group, most of the striated muscles in myocardial tissue of HFpEF group were broken, and the Z line and M line could not be clearly distinguished, some changes such as mitochondrial swelling, membrane thickening, cristae reduction or even disappearance occurred. In the canagliflozin 20 group and canagliflozin 30 group, the arrangement of striated muscles in the myocardial tissue of rats tended to be more regular, and the morphological changes of mitochondria were milder. Prussian blue iron staining results showed that the iron content in myocardial tissue of rats in HFpEF group was higher than that in control group, canagliflozin 20 group and canagliflozin 30 group. Reactive oxygen species staining results showed that the reactive oxygen species content in the myocardial tissue of rats in HFpEF group was higher than that of control group, canagliflozin 20 group and canagliflozin 30 group. Biochemical analysis of myocardial tissue showed that Fe 2+ and malondialdehyde content in myocardial tissue of rats in HFpEF group were higher than those in control group, canagliflozin 20 group and canagliflozin 30 group, while glutathione content was lower (all P<0.05). Western blot and RT-qPCR detection results showed that compared to control group, rats in HFpEF group had higher expression levels of transferrin receptor 1 (protein relative expression level: 1.37±0.16 vs. 0.31±0.12), acyl-CoA synthetase long-chain family member 4 (protein relative expression level: 1.31±0.15 vs. 0.63±0.09) protein and mRNA, and lower expression levels of ferritin heavy chain 1 (protein relative expression level: 0.45±0.08 vs. 1.41±0.15) protein and mRNA (all P<0.05). There was no statistically significant difference in these indicators between canagliflozin 20 group and the canagliflozin 30 group (all P>0.05). There was no significant difference in levels of glutathione peroxidase 4 protein and mRNA expression in myocardial tissue of rats in four groups( P>0.05). Conclusion:Canagliflozin improves cardiac function in HFpEF rats by regulating the ferroptosis mechanism.

Objective:To discuss the differences in serum proline dehydrogenase(ProDH)levels among chronic heart failure(CHF)patients with different ejection fraction types,and to clarify the effect of ProDH levels on cardiac function.Methods:A retrospective analysis of clinical data of 118 CHF patients was conducted.These patients were divided into heart failure with reduced ejection fraction(HFrEF)group(n=39),heart failure with mid-range ejection fraction group(HFmrEF)(n=42),and heart failure with preserved ejection fraction(HFpEF)group(n=37).A total of 45 non-CHF patients hospitalized during the same period were collected as control group.The general data of all the subjects in various groups were collected,and the levels of biochemical indicators and cardiac structure indicators in serum of all the subjects were detected.Spearman correlation analysis and point-biserial correlation analysis were used to analyze the correlation between serum ProDH levels and various biochemical indicators;multivariate Logistic regression analysis was used to analyze the factors influencing HFrEF and HFmrEF.Results:Compared with control group,the usage rate of beta-blockers of the patients in HFpEF group was significantly increased(P<0.05);in HFmrEF group,the percentage of male patients,the usage rate of statins,and the usage rate of beta-blockers were all significantly increased(P<0.05);in HFrEF group,the age and systolic blood pressure(SBP)of the patients were significantly decreased(P<0.05),while the usage rates of statins and beta-blockers of the patients were significantly increased(P<0.05).Compared with HFpEF group,the age of the patients in HFmrEF group was significantly decreased(P<0.05),and the percentage of male patients and the usage rate of statins were significantly increased(P<0.05);the age of the patients in the HFrEF group was significantly decreased(P<0.05),and the usage rate of statins was significantly increased(P<0.05).Compared with HFmrEF group,the SBP of the patients in HFrEF group was significantly decreased(P<0.05).Compared with control group,the serum levels of low-density lipoprotein cholesterol(LDL-c)of the patients in HFpEF and HFmrEF groups were significantly decreased(P<0.05),while the levels of N-terminal pro-brain natriuretic peptide(NT-proBNP)were significantly increased(P<0.05);the serum levels of glomerular filtration rate(GFR)and ProDH of the patients in HFrEF group were significantly decreased(P<0.05),and the levels of fasting blood glucose(FBG)and NT-proBNP were significantly increased(P<0.05).Compared with HFpEF group,the serum hemoglobin(Hb)level of the patients in HFmrEF group was significantly increased(P<0.05);the serum NT-proBNP level of the patients in HFrEF group was significantly increased(P<0.05),while the ProDH level was significantly decreased(P<0.05).Compared with HFmrEF group,the serum NT-proBNP level of the patients in HFrEF group was significantly increased(P<0.05).Compared with control group,the left atrial diameter(LAD)and the ratio of early diastolic mitral inflow velocity to early diastolic mitral annular velocity(E/Em)of the patients in HFpEF,HFmrEF,and HFrEF groups were significantly increased(P<0.05);the left ventricular end-diastolic diameter(LVEDD)of the patients in HFmrEF and HFrEF groups were significantly increased(P<0.05),and the left ventricular ejection fraction(LVEF)were significantly decreased(P<0.05).Compared with HFpEF group,the LVEDD of the patients in HFmrEF and HFrEF groups were significantly increased(P<0.05),and the LVEF were significantly decreased(P<0.05);the LAD of the patients In HFrEF group was significantly increased(P<0.05).Compared with HFmrEF group,the E/Em ratio of the patients in HFrEF group was significantly increased(P<0.05),and the LVEF was significantly decreased(P<0.05).The serum ProDH levels of the patients were negatively correlated with LVEDD(r=-0.210,P=0.007)and positively correlated with LVEF(r=0.220,P=0.005).Male and elevated FBG levels were the risk factors for cardiac function,while the increasing serum GFR and ProDH levels were the protective factors for cardiac function.Conclusion:There are differences in ProDH levels among the CHF patients with different ejection fraction types.The patients with poorer cardiac function have lower serum ProDH levels,and higher ProDH levels may be beneficial for improving the left ventricular systolic function in the CHF patients.