The aim of the study is to identify the effects and underlying mechanisms of visfatin on inflammation and necroptosis in vascular endothelial cells. Human umbilical vein endothelial cells (HUVECs) were stimulated with visfatin or pretreated with Polyinosinic acid (LOX-1 inhibitor). By using the Western blot, RT-PCR, immunocytochemistry, enzyme-linked immunosorbent assay (ELISA), MTT and flow cytometry technique, the occurrence of inflammation and necroptosis in HUVECs were evaluated. Our results showed that 100 ng/mL visfatin significantly increased the mRNA and protein expression of monocyte chemotactic protein 1 (MCP-1) and LOX-1 after 24 hours’ treatment in HUVECs. However, pretreatment with Polyinosinic acid could significantly reduce the expression of MCP-1 compared with visfatin group. Additionally, 100 ng/mL visfatin could induce the production of necrotic features and increase the mRNA expression of BMF (one of the markers of necroptosis), while pretreating with Polyinosinic acid markedly downregulated the mRNA expression of BMF gene and promoted the cell proliferation. These results indicate that visfatin might induce inflammation and necroptosis via LOX-1 in HUVECs, suggesting that visfatin plays a central role in the development of atherosclerosis.
To investigate the value of plasma placental growth factor (PlGF) in percutaneous coronary angioplasty and stent implantation. Methods From May 2006 to March 2007, 61 patients (53 males and 8 females, mean age61 years) and 28 normal controls were included. All patients present with acute chest pain and underwent coronary angiography, the lesion severity of coronary arteries was assessed by Gensini coronary scoring system. Of them, 26 patients having serious coronary lesion underwent (percutaneous transluminal coronary angioplasty, PTCA) and stent implantation. Cardiovascular events were recorded after 30 days. Plasma PlGF was determined by ELISA. Results According to the angiography, the patients could be divided into CAD group (n=45) and Non- CAD group (n=16). Plasma PlGF level in CAD group was significantly higher than that in Non-CAD group and control group [(10.70 ± 0.49) ng/L vs (4.53 ± 0.64) ng/L vs (3.64 ± 0.36) ng/L, P lt; 0.001)], and there was no significant difference between the non-CAD group and control group (P gt; 0.05). A significant positive correlation was found between Gensini coronary score and plasma PlGF level (r=0.918, P lt; 0.01). Moreover, patients with cardiovascular events had a higher PlGF level than those without cardiovascular events after PTCA and stent implantation [(13.98 ± 3.39) ng/L vs (7.25 ± 2.96) ng/L, P lt; 0.01)]. Conclusion PlGF level has diagnostic value in patients with acute chest pain. The measurement of plasma PlGF might be helpful for early diagnosis of coronary artery disease. Patients with higher plasma PlGF level may have more severe coronary lesion. PlGF may be one of predictors for cardiovascular events after PCI.
This study aimed to explore the role of miR-130a-3p in cardiomyocyte hypertrophy and its underlying mechanisms. Pressure-overload induced myocardial hypertrophy mice model was constructed by thoracic aortic constriction (TAC). In vitro, norepinephrine (NE) was used to stimulate neonatal rat cardiomyocytes (NRCMs) and H9c2 rat cardiomyocytes to induce hypertrophic phenotypes. The expression of miR-130a-3p was detected in mice hypertrophic myocardium, hypertrophic NRCMs and H9c2 cells. The mimics and inhibitors of miR-130a-3p were transfected into H9c2 cells to observe the role of miR-130a-3p on the hypertrophic phenotype change of cardiomyocytes separately. Furthermore, whether miR-130a-3p regulated hypertrophic related signaling pathways was explored. The results showed that the expression of miR-130a-3p was significantly decreased in hypertrophic myocardium, hypertrophic NRCMs and H9c2 cells. After transfection of miR-130a-3p mimics, the expression of hypertrophic marker genes, atrial natriuretic peptide (ANP), brain natriuretic peptide (BNP) and β-myosin heavy chain (β-MHC), and the cell surface area were notably down-regulated compared with the control group (mimics N.C. + NE group). But after transfection of miR-130a-3p inhibitor, the expression of ANP, BNP and β-MHC in H9c2 cells increased significantly, and the cell area increased further. By Western blot, it was found that the protein phosphorylation level of Akt and mTOR were down-regulated after over-expression of miR-130a-3p. These results suggest that miR-130a-3p mimics may alleviate the degree of cardiomyocyte hypertrophy, meanwhile its inhibitor can further aggravate cardiomyocyte hypertrophy. Over-expression of miR-130a-3p may attenuate cardiomyocytes hypertrophy by affecting the Akt pathway.
Calnexin is a lectin-like molecular chaperone protein on the endoplasmic reticulum, mediating unfolded protein responses, the endoplasmic reticulum Ca2+ homeostasis, and Ca2+ signals conduction. In recent years, studies have found that calnexin plays a key role in the heart diseases. This study aims to explore the role of calnexin in the activation of cardiac fibroblasts. A transverse aortic constriction (TAC) mouse model was established to observe the activation of cardiac fibroblasts in vivo, and the in vitro cardiac fibroblasts activation model was established by transforming growth factor β1 (TGFβ1) stimulation. The adenovirus was respectively used to gene overexpression and silencing calnexin in cardiac fibroblasts to elucidate the relationship between calnexin and cardiac fibroblasts activation, as well as the possible underlying mechanism. We confirmed the establishment of TAC model by echocardiography, hematoxylin-eosin, Masson, and Sirius red staining, and detecting the expression of cardiac fibrosis markers in cardiac tissues. After TGFβ1 stimulation, markers of the activation of cardiac fibroblast, and proliferation and migration of cardiac fibroblast were detected by quantitative PCR, Western blot, EdU assay, and wound healing assay respectively. The results showed that the calnexin expression was reduced in both the TAC mice model and the activated cardiac fibroblasts. The overexpression of calnexin relieved cardiac fibroblasts activation, in contrast, the silencing of calnexin promoted cardiac fibroblasts activation. Furthermore, we found that the endoplasmic reticulum stress was activated during cardiac fibroblasts activation, and endoplasmic reticulum stress was relieved after overexpression of calnexin. Conversely, after the silencing of calnexin, endoplasmic reticulum stress was further aggravated, accompanying with the activation of cardiac fibroblasts. Our data suggest that the overexpression of calnexin may prevent cardiac fibroblasts against activation by alleviating endoplasmic reticulum stress.
Long non-coding RNA (lncRNA) Dnm3os plays a critical role in peritendinous fibrosis and pulmonary fibrosis, but its role in the process of cardiac fibrosis is still unclear. Therefore, we carried out study by using the myocardial fibrotic tissues obtained by thoracic aortic constriction (TAC) in an early study of our group, and the in vitro cardiac fibroblast activation model induced by transforming growth factor-β1 (TGF-β1). Quantitative real-time polymerase chain reaction (RT-qPCR), Western blot, and collagen gel contraction test were used to identify the changes of activation phenotype and the expression of Dnm3os in cardiac fibroblasts. Small interfering RNA was used to silence Dnm3os to explore its role in the activation of cardiac fibroblasts. The results showed that the expression of Dnm3os was increased significantly in myocardial fibrotic tissues and in the activated cardiac fibroblasts. And the activation of cardiac fibroblasts could be alleviated by Dnm3os silencing. Furthermore, the TGF-β1/Smad2/3 pathway was activated during the process of cardiac fibroblasts activation, while was inhibited after silencing Dnm3os. The results suggest that Dnm3os silencing may affect the process of cardiac fibroblast activation by inhibiting TGF-β1/Smad2/3 signal pathway. Therefore, interfering with the expression of lncRNA Dnm3os may be a potential target for the treatment of cardiac fibrosis.
Objective To study the factors that affect the prognosis of status epilepticus (SE) and to improve the understanding of clinicians. Methods A retrospective analysis of 57 patients with SE witch from the General Hospital of Ningxia Medical University and Cardio-cerebrovascular Disease Hospital were carried out to collect their clinical data. The data were analyzed by SPSS 17.0 software. The prognosis of the patients was assessed by the Status epilepticus severity score (STESS) scale. Results A total of 57 patients were included, 53 cases improved, 4 cases were automatically discharged. Telephone follow-up showed that 4 cases of automatic discharge were dead. The mortality rate of SE was 7.02%. The most common cause of SE was acute cerebrovascular disease (17.54%), followed by intracranial infection (10.53%); The most common incidence were the occasional medication, self-medication, withdrawal (15.79%). Age, state of consciousness and concurrent infection were associated with prognosis (improvement/death) (P<0.05). STESS score of 0 to 2 points were 45 patients, all improved; score of 3 to 5 points were 12 patients, 8 patients improved, 4 patients died. There were significant differences in the prognosis between the two groups (P<0.05). Conclusions Age, state of consciousness, concurrent infection were related to prognosis, more than 65 years, the state of consciousness for the sleeping or coma had the poor prognosis. STESS scale can predict the prognosis of patients effectively.