Cardioprotective anti-inflammatory activities of Artemisia lactiflora Wall. ex DC. extract and fractions in a rat cardiomyoblast (H9c2) model of inflammatory sepsis

Context: Sepsis is a progressive inflammatory response commonly caused by gram-negative bacteria via activation of the NF-κB-dependent pathway. Severe sepsis can cause cardiac dysfunction and increases the risk of death. Artemisia lactiflora is a Chinese medicinal plant that contains phenolics and flavonoids with probable anti-inflammatory activities. Aims: To investigate the anti-inflammatory activities of A. lactiflora leaves extract in a cardiac sepsis model. Methods: A. lactiflora leaves were extracted and fractioned using solvents of different polarities. The total phenolic and flavonoid content was quantified. The antioxidants were quantified in vitro using DPPH and ABTS radicals. The cytotoxicity of the extract and fractions was evaluated and calculated using the neutral red assay and curve-fitting analysis in cardiomyoblasts (H9c2). The anti-inflammatory activities of A. lactiflora were observed by treating H9c2 with extract and fractions in the presence of Escherichia coli. Expression of pro-inflammatory genes (TNF and IL6) and secretion of pro-inflammatory cytokines (TNF-α and IL-6) were measured by RT-qPCR and ELISA. Results: Extract and fractions of A. lactiflora contained noticeably high quantities of phenolics and flavonoids with considerably high anti-DPPH and anti-ABTS activities. Co-treatment with A. lactiflora extract and all fractions significantly downregulated the expression of TNF and IL6 as well as decreased the secretion of TNF-α and IL-6 (p<0.0001) compared with E. coli-stimulated cardiomyoblasts. The butanol fraction had the highest potency in reducing pro-inflammatory cytokine secretions in cardiomyoblasts under in vitro sepsis conditions. Conclusions: A. lactiflora leaf extract demonstrated therapeutic potential in a cardiac sepsis model by alleviating the inflammatory responses. This plant can potentially be developed as an alternative medicine for inflammation and sepsis

Combination of metformin and p38 MAPK inhibitor, SB203580, reduced myocardial ischemia/reperfusion injury in non‑obese type 2 diabetic Goto‑Kakizaki rats

International audienceDiabetic cardiomyopathy, especially myocardial ischemia reperfusion (I/R) injury, is a major cause of morbidity and mortality in type 2 diabetic patients. The increasing of basal p38 MAP Kinase (p38 MAPK) activation is a major factor that aggravates cardiac death on diabetic cardiomyopathy. In addition, metformin also shows cardio-protective effects on myocardial ischemia/reperfusion injury. In this study, we investigated the effect of the combination between metformin and p38 MAPK inhibitor (SB203580) in diabetic rats subjected to I/R injury. H9c2 cells were induced into a hyperglycemic condition and treated with metformin, SB203580 or the combination of metformin and SB203580. In addition, cells in both the presence and absence of drug treatment were subjected to simulated ischemia/reperfusion injury. Cell viability and cellular reactive oxygen species (ROS) were determined. Moreover, the Goto-Kakizaki (GK) rats were treated with metformin, SB203580, and the combination of metformin and SB203580 for 4 weeks. Diabetic parameters and cardiac functions were assessed. Finally, rat hearts were induced ischemia/reperfusion injury for the purpose of infarct size analysis and determination of signal transduction. A high-glucose condition did not reduce cell viability but significantly increased ROS production and significantly decreased cell viability after induced sI/R. Treatment using drugs was shown to reduce ROS generation and cardiac cell death. The GK rats displayed diabetic phenotype by increasing diabetic parameters and these parameters were significantly decreased when treated with drugs. Treatment with metformin or SB203580 could significantly reduce the infarct size. Interestingly, the combination of metformin and SB203580 could enhance cardio-protective ability. Myocardial I/R injury significantly increased p38 MAPK phosphorylation, Bax/Bcl-2 ratio and caspase-3 level. Treatment with drugs significantly decreased the p38 MAPK phosphorylation, Bax/Bcl-2 ratio, caspase-3 level and increased Akt phosphorylation. In conclusion, using the combination of metformin and SB203580 shows positive cardio-protective effects on diabetic ischemic cardiomyopathy