Qifu Yixin Formula Improves Heart Failure by Enhancing ß-Arrestin2 Mediated the SUMOylation of SERCA2a.

Purpose: This study aimed to elucidate the protective mechanism of Traditional Chinese Medicine (TCM) Qifu Yixin formula (QFYXF) to improve heart failure (HF) by promoting ß-arrestin2 (ß-arr2)-mediated SERCA2a SUMOylation. Materials and Methods: The transverse aortic constriction (TAC)-induced HF mice were treated with QFYXF or carvedilol for 8 weeks. ß-arr2-KO mice and their littermate wild-type (WT) mice were used as controls. Neonatal rat cardiomyocytes (NRCMs) were used in vitro. Cardiac function was evaluated by echocardiography and serum NT-proBNP. Myocardial hypertrophy and myocardial fibrosis were assessed by histological staining. ß-arr2, SERCA2a, SUMO1, PLB and p-PLB expressions were detected by Western blotting, immunofluorescence and immunohistochemistry. SERCA2a SUMOylation was detected by Co-IP. The molecular docking method was used to predict the binding ability of the main active components of QFYXF to ß-arr2, SERCA2a, and SUMO1, and the binding degree of SERCA2a to SUMO1 protein. Results: The HF model was constructed 8 weeks after TAC. QFYXF ameliorated cardiac function, inhibiting myocardial hypertrophy and fibrosis. QFYXF promoted SERCA2a expression and SERCA2a SUMOylation. Further investigation showed that QFYXF promoted ß-arr2 expression, whereas Barbadin (ß-arr2 inhibitor) or ß-arr2-KO reduced SERCA2a SUMOylation and attenuated the protective effect of QFYXF improved HF. Molecular docking showed that the main active components of QFYXF had good binding activities with ß-arr2, SERCA2a, and SUMO1, and SERCA2a had a high binding degree with SUMO1 protein. Conclusion: QFYXF improves HF by promoting ß-arr2 mediated SERCA2a SUMOylation and increasing SERCA2a expression.

Investigating the Mechanism of Chinese Medicine Formula AACO Against Chronic Heart Failure by Network Pharmacology Analysis and Experimental Validation.

Background: A traditional Chinese medicine (TCM) formula, containing Astragalus membranaceus (Fisch.) Bunge, Aconitum wilsonii Stapf ex Veitch, Curcuma longa L., and Radix ophiopogonis (AACO), has therapeutic value for the treatment of chronic heart failure (CHF). Objective: This study intends to explore the pharmacological mechanism underlying the activity of the AACO formula against CHF. Materials and Methods: Using the TCM Systems Pharmacology database and Bioinformatics Analysis Tool for Molecular Mechanism of TCM, the active ingredients contained in the herbs of the AACO formula were screened. Meanwhile, the target genes related to these active ingredients were identified and genes correlated with CHF were screened. Protein-protein interaction networks were built to elucidate the relationships between the AACO formula and CHF. Gene ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) signal pathway enrichment analysis were carried out using the DAVID database. A "drug-component-target-disease" network was constructed with Cytoscape 3.7.0. The therapeutic effect of the AACO formula was proven by hemodynamic study, echocardiography evaluation, and histological analysis in transverse aortic constriction-induced CHF mice and was validated in vitro. Results: A total of 105 active ingredients and 1026 related targets were screened and identified, and 240 related targets overlapping with CHF were selected. According to GO analysis, the enriched genes participated in gene expression and cardiac contraction regulation by Ca2+ regulation. From KEGG analysis, the calcium axis was identified as one of the main mechanisms through which the AACO formula exerts an anti-CHF effect. AACO was validated to significantly improve cardiac diastolic and systolic functions in vivo via an increase in the rate of Ca2+ reuptake of the myocardial sarcoplasmic reticulum and improved myocardial contractility in vitro. Conclusions: Network pharmacology is a convenient method to study the complex pharmacological mechanisms of TCM. The calcium axis likely participates in the anti-CHF mechanism of AACO.

LuQi formula attenuates Cardiomyocyte ferroptosis via activating Nrf2/GPX4 signaling axis in heart failure.

BACKGROUND: The terminal stage of all cardiovascular diseases typically culminates in heart failure (HF), with no effective intervention available to halt its progression. LuQi formula (LQF) has been employed in clinical for numerous years to significantly ameliorate cardiac function in HF patients. Nevertheless, the underlying mechanism of LQF's efficacy remains inadequately comprehended. Cardiomyocyte ferroptosis has served as a pathogenic mechanism in HF. The goal of the current experiment was to ascertain whether LQF ameliorates HF by preventing cardiomyocyte ferroptosis and to elucidate the intrinsic mechanism involved. PURPOSE: This research objective is to investigate the impact and underlying mechanism of LQF attenuating cardiomyocyte ferroptosis in heart failure. METHODS: Transverse aortic constriction (TAC) was performed to construct the HF mouse model. Neonatal rat cardiomyocytes (NRCMs) were subjected to in vitro experiments. High-performance liquid chromatography (HPLC) identified the bioactive compounds in LQF. Transcriptomic and quantitative proteomic analyses revealed the potential targets of LQF anti-HF. Specifically, histological staining evaluated cardiac hypertrophy and fibrosis. Transmission electron microscopy (TEM) observed mitochondrial morphology. The content of Fe2+, ROS, MDA, GSH, and GSSH was detected using kits. Molecular docking evaluated the binding activities between essential active ingredients of LQF and critical proteins of cardiomyocyte ferroptosis. Mechanistically, the expression levels of Nrf2, Keap1, HO-1, SLC7A11, and GPX4 were evaluated using qPCR, Western blot (WB), or immunohistochemical staining. RESULTS: The primary nine active ingredients in LQF were detected. Transcriptomic and proteomic analyses demonstrated that LQF may ameliorate HF by preventing cardiomyocyte ferroptosis. Histomorphometric analyses revealed that LQF attenuates myocardial hypertrophy and fibrosis. TEM revealed that LQF diminished mitochondrial shrinkage and increased membrane density in myocardial tissue. Additionally, LQF diminished reactive oxygen species (ROS) generation in cardiomyocytes and suppressed cardiomyocyte ferroptosis. Furthermore, the molecular docking technique revealed that the primary active ingredients of LQF had suitable binding activities with Nrf2, GPX4, and SLC7A11. Western analysis further verified that LQF activated the Nrf2/GPX4 signaling axis. decreased SLC7A11 and HO-1 expression. CONCLUSIONS: These results demonstrated that LQF prevents cardiomyocyte ferroptosis via activating Nrf2/GPX4 signaling axis and suppressing SLC7A11 and HO-1 expression. Concurrently, it contributed to elucidating the intrinsic mechanism of LQF and provided a scientific rationale for its development as a novel cardiovascular therapeutic drug.

Moxibustion improved the effect of fecal microbiota transplantation donor to dextran sulfate sodium-induced colitis in mice.

Fecal microbiota transplantation (FMT) is beneficial for several gastrointestinal diseases because it alters the intestinal microbiota of recipients. The efficacy of FMT is related to the microbial structure and composition of the donor. Mild moxibustion is a non-invasive and safe traditional Chinese therapy that can regulate the gut microbiota. In this study, we investigated whether moxibustion improved the efficacy of FMT in donors using a dextran sulfate sodium (DSS)-induced colitis mouse model. Normal mice were treated with mild moxibustion at acupoints ST25 and ST36 for 7 days. DSS (2%) was administered for 7 days to induce colitis. FMT was performed on Day 8 and lasted for 7 days. The effect of FMT on mice with DSS was observed on Day 21. Using hematoxylin and eosin staining and immunofluorescence, we analyzed the pathology and cell proliferation after FMT in DSS mice. In addition, using 16 S rDNA sequencing analysis, we investigated the gut microbiota of mice. The results indicated that moxibustion altered the colonic microbial community and increased the relative abundance of specific bacteria without changes in morphology and physiological function in normal mice. FMT using donors with moxibustion reduced body weight loss, inflammation, abnormal microbial community structure, and the relative abundance of some bacteria. These results provide potential strategies for the safe and targeted improvement of FMT donors.

4cRNA NEAT1 Sponge Adsorption of miR-378 Modulates Activity of Lipopolysaccharide-treated Articular Chondrocytes and Influences the Pathological Development of Osteoarthritis.

Context: Osteoarthritis (OA) is a chronic joint disease that can eventually lead to degeneration, fibrosis, fractures, and defects of the articular cartilage. Long non-coding RNA (lncRNA) is a key substance in many processes, such as epigenetic regulation and cell-cycle and cell-differentiation modulation, and its relationship with OA has been repeatedly verified. Objective: The study intended to clarify the influence of lncRNA nuclear enriched abundant transcript 1 (NEAT1), lncRNA NEAT1, on lipopolysaccharide (LPS)-induced OA chondrocytes through sponge adsorption of microRNA-378 (miR-378) and to provide novel insights into future diagnosis and treatment of OA. Design: The research team performed an animal study. Setting: The study took place in the Department of Rehabilitation Medicine at Linyi People's Hospital in Linyi, Shangdong, China. Animals: The study's animals were 10 Sprague Dawley (SD) rats, 3-5 days old and 10-15 g in weight, of the specific-pathogen-free (SPF) grade. Intervention: The rat chondrocytes for the positive control group (the model group) were treated with 500 ng/mL of LPS to induce OA. Chondrocytes treated with the same amount of normal saline were used as the negative control group. The chondrocytes of the LPS-induced rats were into six groups: (1) a positive control group transfected with NEAT1-interfering RNA, the sh-NEAT1 group; (2) a negative control group not transfected with NEAT1-interfering RNA, the NEAT1 empty vector (NC-NEAT1) group; (3) an intervention group co-transfected with NEAT1 interfering RNA and the miR-378 inhibitor sequence (Inh-miR-378 the sh-NEAT1+ Inh-miR-378 group; (4) a negative control group transfected with NEAT1 interfering RNA but not transfected with the miR-378 inhibitor sequence, the sh-NEAT1+ miR-378 negative control (NC-miR-378) group; (5) a negative control group transfected with the miR-378 inhibitor sequence but not transfected with NEAT1 interfering RNA, the NEAT1 empty vector (NC-NEAT1) + Inh-miR-378 group; (6) a negative control group not transfected with either NEAT1 interfering RNA or the miR-378 inhibitor sequence, the NC-NEAT1 + NC-miR-378 group. Outcome Measures: An OA-chondrocyte model was induced by LPS and measurements of NEAT1 and miR-378 expression were made by real-time quantitative reverse transcription (qRT)- polymerase chain reaction (PCR). Then, small NEAT1-interfering RNA (sh-NEAT1), empty vector NEAT1 (NC-NEAT1), inhibitor-sequence-miR-378 (Inh-miR-378), and negative-control-miR-378 (NC-miR-378) were transfected into cells, and cell viability and apoptosis rate were measured. Finally, the study verified the relationship between NEAT1 and miR-378. Results: Compared to the control group, NEAT1 was significantly elevated in the model group, and its miR-378 was significantly decreased. Silencing NEAT1 can enhance OA-chondrocyte activity and decrease apoptosis. When NEAT1 and miR-378 were inhibited together, as shown fort the NC-NEAT1 + NC-miR-378 group, NEAT1 expression, as well as the multiplication and apoptosis ability of the OA-model cells, were the same as those of cells transfected with an empty vector, the NC-NEAT1 group. Also, the NEAT1 + NC-miR-378 group's cell activity was lower than that of the sh-NEAT1+NC-miR-378 group but higher than that of the NC-NEAT1 + Inh-miR-378 group. Finally, higher fluorescence activity occurred for NEAT1-mutant type (MUT) transfected with Inh-miR-378. Conclusions: NEAT1, which is highly expressed in OA, mediates LPS-induced OA-chondrocyte activity through sponge adsorption of miR-378.

Factors contributing to the high prevalence of multidrug-resistant tuberculosis among previously treated patients: a case-control study from China.

SETTING: Multidrug-resistant tuberculosis (MDR-TB) has emerged as a serious global public health problem. In China, the risk factors for MDR-TB have not been systematically evaluated. OBJECTIVE: To identify risk factors associated with MDR-TB among previously treated patients in China. DESIGN: A case-control study was carried out. Cases were selected from previously treated MDR-TB patients who were resistant to both isoniazid and rifampin, and controls were selected from previously treated TB patients who were sensitive to isoniazid and rifampin (non-MDR-TB). Information was collected from the registration database and a structured questionnaire. RESULTS: A total of 61 cases and 50 controls were recruited. A multivariate analysis showed that the family annual per-capita income ≤7,000 Yuan (odds ratio [OR]=3.238; 95% confidence interval [CI]: 1.270-8.252), no history of fixed dose combinations (FDCs) in anti-TB treatment (OR=4.027; 95% CI: 1.457-11.129), and adverse reactions in the course of TB treatment (OR=3.568; 95% CI: 1.402-9.085) were independent predictors of MDR-TB. Moreover, among the TB patients who had adverse reactions, quitting the treatment was shown as a risk factor for MDR-TB (p=0.009). CONCLUSION: In the control of MDR-TB among previously treated patients, lower socioeconomic groups, the expanding use of FDCs, and improving adherence to treatment by implementing Directly Observed Therapy Short Course-Plus (DOTS-Plus), strictly should become a priority that requires strong commitment and collaboration among health organizations.

Increased plasma exposures of five protoberberine alkaloids from Coptidis Rhizoma in streptozotocin-induced diabetic rats: is P-GP involved?

Our previous study showed a higher exposure of berberine, palmatine, coptisine, epiberberine and jatrorrhizine in 6-week streptozotocin (STZ)-induced diabetic rats, after oral administration of Coptidis Rhizoma extract. The aim of the present study was to investigate whether the function and expression of intestinal P-glycoprotein (P-GP) was downregulated in STZ-induced diabetic rats and if the impairment of P-GP function and expression contributed to the exposure increase of the five protoberberine alkaloids. Plasma concentration-time profiles of the drugs in the portal vein were obtained after oral administration of Coptidis Rhizoma extract. The effective permeability of the drug across duodenum and ileum were measured using in situ single-pass intestine perfusion. P-GP function in the rat intestine was assessed by measuring the absorption of rhodamine 123 (Rho123). P-GP levels were evaluated using Western blots. It was found that the C(max) and AUC(0-8) values of five alkaloids in the portal vein of diabetic rats were significantly higher than those in the control rats. Diabetic rats also exhibitd a higher level of Rho123 in the portal vein, which showed impairment of P-GP function. A higher effective permeability of the tested drug was found in the duodenum of diabetic rats using in situ single-pass intestine perfusion, indicating that berberine and Rho123 transported more easily across the intestinal barrier of diabetic rats. A lower level of P-GP protein was found in the duodenum, jejunum and ileum of the diabetic rats as compared with age-matched control rats. All these results suggested that the function and expression of P-GP were impaired in the intestine of STZ-induced diabetic rats which, at least partly, contributed to the exposure increase of the five protoberberine alkaloids.