Zonisamide attenuates pressure overload-induced myocardial hypertrophy in mice through proteasome inhibition.

Myocardial hypertrophy is a pathological thickening of the myocardium which ultimately results in heart failure. We previously reported that zonisamide, an antiepileptic drug, attenuated pressure overload-caused myocardial hypertrophy and diabetic cardiomyopathy in murine models. In addition, we have found that the inhibition of proteasome activates glycogen synthesis kinase 3 (GSK-3) thus alleviates myocardial hypertrophy, which is an important anti-hypertrophic strategy. In this study, we investigated whether zonisamide prevented pressure overload-caused myocardial hypertrophy through suppressing proteasome. Pressure overload-caused myocardial hypertrophy was induced in mice by trans-aortic constriction (TAC) surgery. Two days after the surgery, the mice were administered zonisamide (10, 20, 40 mg·kg-1·d-1, i.g.) for four weeks. We showed that zonisamide administration significantly mitigated impaired cardiac function. Furthermore, zonisamide administration significantly inhibited proteasome activity as well as the expression levels of proteasome subunit beta types (PSMB) of the 20 S proteasome (PSMB1, PSMB2 and PSMB5) and proteasome-regulated particles (RPT) of the 19 S proteasome (RPT1, RPT4) in heart tissues of TAC mice. In primary neonatal rat cardiomyocytes (NRCMs), zonisamide (0.3 µM) prevented myocardial hypertrophy triggered by angiotensin II (Ang II), and significantly inhibited proteasome activity, proteasome subunits and proteasome-regulated particles. In Ang II-treated NRCMs, we found that 18α-glycyrrhetinic acid (18α-GA, 2 mg/ml), a proteasome inducer, eliminated the protective effects of zonisamide against myocardial hypertrophy and proteasome. Moreover, zonisamide treatment activated GSK-3 through inhibiting the phosphorylated AKT (protein kinase B, PKB) and phosphorylated liver kinase B1/AMP-activated protein kinase (LKB1/AMPKα), the upstream of GSK-3. Zonisamide treatment also inhibited GSK-3's downstream signaling proteins, including extracellular signal-regulated kinase (ERK) and GATA binding protein 4 (GATA4), both being the hypertrophic factors. Collectively, this study highlights the potential of zonisamide as a new therapeutic agent for myocardial hypertrophy, as it shows potent anti-hypertrophic potential through the suppression of proteasome.

Study of the Mechanism of Astragali Radix in Treating Type 2 Diabetes Mellitus and Its Renal Protection Based on Enzyme Activity, Network Pharmacology, and Experimental Verification.

Astragali Radix (AR) is a common Chinese medicine and food. This article aims to reveal the active role of AR in treating Type 2 diabetes mellitus (T2DM) and its renal protective mechanism. The hypoglycemic active fraction was screened by α-glucosidase and identified by UPLC-QE-Orbitrap-MS spectrometry. The targets and KEGG pathway were determined through the application of network pharmacology methodology. Molecular docking and molecular dynamics simulation technology were used for virtual verification. Subsequently, a mouse model of T2DM was established, and the blood glucose and renal function indexes of the mice after administration were analyzed to further prove the pharmacodynamic effect and mechanism of AR in the treatment of T2DM. HA was determined as the best hypoglycemic active fraction by the α-glucosidase method, with a total of 23 compounds identified. The main active components, such as calycoside-7-O-ß-D-glucoside, methylnisoline, and formononetin, were revealed by network pharmacology. In addition, the core targets and the pathway have also been determined. Molecular docking and molecular dynamics simulation techniques have verified that components and targets can be well combined. In vivo studies have shown that AR can reduce blood sugar levels in model mice, enhance the anti-inflammatory and antioxidant activities of kidney tissue, and alleviate kidney damage in mice. And it also has regulatory effects on proteins such as RAGE, PI3K, and AKT. AR has a good therapeutic effect on T2DM and can repair disease-induced renal injury by regulating the RAGE/PI3K/Akt signaling pathway. This study provides ideas for the development of new drugs or dietary interventions for the treatment of T2DM.

A novel onco-cardiological mouse model of lung cancer-induced cardiac dysfunction and its application in identifying potential roles of tRNA-derived small RNAs.

An increasing body of research suggests cancer-induced cardiovascular diseases, leading to the appearance of an interdisciplinary study known as onco-cardiology. Lung cancer has the highest incidence and mortality. Cardiac dysfunction constitutes a major cause of death in lung cancer patients. However, its mechanism has not been elucidated because suitable animal models that adequately mimic clinical features are lacking. Here, we established a novel chemically induced lung cancer mouse model using benzo[a]pyrene and urethane to recapitulate the general characteristics of cardiac dysfunction caused by lung cancer, the cardiac disorders in the context of the progression of lung cancer were evaluated using echocardiographic and histological approaches. The pathological changes included myocardial ischaemia, pericarditis, cardiac pre-cachexia, and pulmonary artery hypertension. We performed sequencing to detect the tRNA-derived fragments and tRNA-derived stress-induced RNAs (tRFs/tiRNAs) expressions in mouse heart tissue. 22 upregulated and 16 downregulated tRFs/tiRNAs were identified. Subsequently, the top 10 significant results of Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analysis were presented. The in vitro model was established by exposing neonatal rat cardiomyocytes and myocardial fibroblasts to lung tumour cell-conditioned medium, respectively. Western blotting revealed significant changes in cardiac failure markers (atrial natriuretic peptide and α-myosin heavy chain) and cardiac fibrosis markers (Collagen-1 and Collagen-3). Our model adequately reflects the pathological features of lung cancer-induced cardiac dysfunction. Furthermore, the altered tRF/tiRNA profiles showed great promise as novel targets for therapies. These results might pave the way for research on therapeutic targets in onco-cardiology.

[ClinicaAnalysis of osteosclerotic protein expression and bacterial distribution in periodontitis patients at different stages].

PURPOSE: To investigate the osteosclerin level and bacterial distribution in periodontitis patients at different stages, and to analyze the correlation between osteosclerin and the parameters of conventional periodontal examination. METHODS: Patients with periodontitis admitted to Guangzhou Huadu Maternal and Child Health Hospital from March 2017 to June 2019 were selected and divided into stage Ⅱ group (n=27), stage Ⅲ group (n=42) and stage Ⅳ group (n=22) according to the severity of periodontitis; meanwhile, 30 healthy individuals underwent physical examination in our hospital during the same period were selected as control group. Gingival crevicular fluid and plaque at buccal and lingual sites were collected for bacterial culture. The expression of osteosclerotin in gingival crevicular fluid was detected by enzyme-linked immunosorbent assay. The data were processed by SPSS 23.0 software package. Spearman correlation analysis was used to analyze the correlation between BI grade and osteosclerin, and correlation between PD, CAL and osteosclerin was determined by Pearson analysis. RESULTS: The mean PD and mean CAL of patients in stage Ⅱ group before and after treatment were significantly smaller than those in stage Ⅲ and Ⅳ group (P<0.05). The mean CAL of stage Ⅳ group before treatment was significantly greater than that of stage Ⅲ group (P<0.05). After treatment, the mean PD and mean CAL of three groups were all significantly smaller than those before treatment (P<0.05). The mean PD in stage Ⅲ group was significantly lower than that in stage Ⅳ group after treatment (P<0.05). Before treatment, the proportion of BI grade 2 in stage Ⅱ group was significantly higher than that in stage Ⅲ and Ⅳ group (85.19%, 19.05%, 18.18%, P<0.05). Before treatment, the proportion of BI grade 3 in stage Ⅲ group was significantly higher than that in stage Ⅱ group (64.29%, 14.81%, P<0.05). Before the treatment, the expression of osteosclerosis protein in stage Ⅱ group was significantly lower than that in stage Ⅲ and Ⅳ group (P<0.05). The levels of osteosclerin expression of three groups after treatment were all significantly lower than those before treatment (P<0.05). The expression of osteosclerosis protein in stage Ⅱ group was significantly lower than that in stage Ⅲ and Ⅳ group after treatment (P<0.05). PD, CAL and BI of patients with different stages of periodontitis were positively correlated with osclerosin in gingival crevicular fluid before and after treatment (P<0.05). The number of bacteria detected in stage Ⅳ group was significantly higher than that in stage Ⅲ group and stage Ⅱ group. The main bacteria in each group were anaerobic bacteria. The dominant bacteria were Porphyromonas gingivalis, Prevotella intermedia, Actinobacillus actinomycetes, Fusobacterium nucleatum, and Prevotella melaninogenicus. CONCLUSIONS: The expression level of osteosclerosin is closely related to PD, CAL and BI grades in patients with periodontitis, and bacterial colonization levels in gingival crevicular fluid and dental plaque in patients with periodontitis at different stages are different. Detection of osclerosin level and identification of periodontal microorganism culture have high clinical value in clinical diagnosis of periodontitis severity and can provide reference for selection of subsequent treatment plan.

[Study on clinical efficacy and patients' satisfaction of MTA apical barrier technique in treatment of young permanent teeth with periapical inflammation].

PURPOSE: To compare the clinical efficacy of MTA apical barrier technique and Vitapex apexification in treatment of young permanent teeth with periapical inflammation, and to evaluate the satisfaction of patients. METHODS: Seventy-five cases of young permanent teeth with periapical inflammation were randomly divided into control group (n=37) and experimental group (n=38). Patients in the control group were treated with Vitapex apexification, while patients in the experimental group were treated with MTA apical barrier technique. The clinical efficacy of the two groups was compared at 3, 6, 9 months and 1 year after treatment, and the average treatment time and average treatment period were compared between 2 groups. The difference of patients' satisfaction with medical environment, health care service, late health care guidance, treatment cost, treatment period and treatment effect were compared between 2 groups. The clinical efficacy, treatment times and period, satisfaction of 2 groups were recorded and analyzed by using SPSS 19.0 software package. RESULTS: At 3 month and 6 month of revisit, the clinical efficacy of the experimental group was better than the control group, but there was no significant difference between 2 groups (P>0.05). At 9 months and 1 year of revisit, the total efficiency of the experimental group was significantly better than the control group (78.38%:94.74%, P=0.037；75.68%∶97.37%, P=0.006). The treatment time and treatment period of the experimental group were significantly lower than the control group (P<0.05), the values were (3.24±0.39) times, (0.68±0.23) months and (7.78±0.65) times, (8.24±2.95) months. Patients' satisfaction with medical treatment environment, health care service, late health care guidance and treatment period was not significant different between 2 groups (P>0.05). However, patients' satisfaction with treatment cost and treatment effect in the experimental group was significantly higher than the control group (P<0.05). CONCLUSIONS: MTA apical barrier technique has better clinical efficacy, less treatment time, shorter treatment period and higher satisfaction than Vitapex apexification. It is suitable for clinical application.