Combination Therapy of Amlodipine and Atorvastatin Has More Beneficial Vascular Effects Than Monotherapy in Salt-Sensitive Hypertension

BACKGROUND Current treatment for the secondary prevention of cardiovascular diseases frequently involves the prescription of several combination therapies, particularly antihypertensive medications and HMG-CoA reductase inhibitor. We have previously shown that in salt-sensitive hypertension either a statin or the calcium channel blocker amlodipine (Aml) have vasoprotective effects. Here, we investigated in aortas from Dahl salt-sensitive (DS) rats the effects of Aml, the statin atorvastatin (AT), and their combination on endothelial function, superoxide (O2−) production, and the expression of endothelial nitric oxide synthase (eNOS), chemokine monocyte chemoattractant protein-1 (MCP-1), and lectin-like oxidized LDL receptor-1 (LOX-1). METHODS Groups of DS rats were fed either normal-salt (NS, 0.5% NaCl) or high-salt (HS, 4% NaCl) diet or a HS diet with AT (15mg/kg/day), Aml (5mg/kg/day) or combination of AT/Aml for 6 weeks. RESULTS Rats on the HS diet developed hypertension, aortic hypertrophy, accompanied by increased plasma C-reactive protein (CRP), aortic O2−, MCP-1 (80%), and LOX-1 (55%) expression and reduced eNOS and endothelial-dependent relaxation to acetylcholine (EDR). Aml reduced systolic blood pressure (SBP), aortic hypertrophy, plasma CRP, vascular O2−, and MCP-1 expression and improved eNOS and EDR. AT reduced aortic hypertrophy and plasma CRP, improved EDR, and normalized vascular O2−, eNOS, and proinflammatory gene expression with mild reduction in SBP. Combination therapy further reduced the SBP and normalized aortic hypertrophy, EDR, and plasma CRP. CONCLUSIONS The combination therapy of Aml/AT has an additive beneficial effect on the vasculature. These novel findings may provide scientific basis for the combination therapy of statins with antihypertensive agents to reduce and prevent cardiovascular diseases

Novel three-dimensional biochip pulmonary sarcoidosis model.

Sarcoidosis is a multi-system disorder of granulomatous inflammation which most commonly affects the lungs. Its etiology and pathogenesis are not well defined in part due to the lack of reliable modeling. Here, we present the development of an in vitro three-dimensional lung-on-chip biochip designed to mimic granuloma formation. A lung on chip fluidic macrodevice was developed and added to our previously developed a lung-on-membrane model (LOMM). Granulomas were cultured from blood samples of patients with sarcoidosis and then inserted in the air-lung-interface of the microchip to create a three-dimensional biochip pulmonary sarcoidosis model (3D BSGM). Cytokines were measured after 48 hours. ELISA testing was performed to measure cytokine response difference between LOMM with 3D BSGM. There were statistically significant differences in IL-1ß (P = 0.001953), IL-6 (P = 0.001953), GM-CSF (P = 0.001953), and INF-γ expressions (P = 0.09375) between two groups. The current model represents the first 3D biochip sarcoidosis model created by adding a microfluidics system to a dual-chambered lung on membrane model and introducing developed sarcoid-granuloma to its air-lung-interface

Abstract 231: Role of SDF-1α-induced Double Reciprocal E-Selectin/Ligand Pairs in Mediating Recruitment of Endothelial Progenitor Cells to Ischemic Wounds

Introduction: We previously reported that stromal cell-derived factor-1α (SDF-1α)-induced endothelial progenitor cells (EPC) homing is enhanced by up-regulation of E-selectin (E-sel) on luminal endothelial cells (EC) lining capillaries, locally within wound tissues. We now investigate how elevated SDF-1α in wounds achieves a remote effect on bone marrow-derived (BMD) circulating EPC. Methods: SDF-1α-induced cell adhesion molecules in human EPC and human microvascular endothelial cells (HMVEC) were profiled using PCR-Array and validated by immunoblot. Induction of E-sel expression on murine BMD- and circulating EPC by SDF-1α was examined by flow cytometry. The involvement of E-sel/ligand in mediating SDF-1α-induced EPC-EC interaction and EPC transendothelial migration was studied using in vitro EPC-EC binding assay. The role of SDF-1α-induced E-sel in regulating EPC homing, wound neovascularization and healing were furthered studied by testing EPC from ROSA26-LacZ+/E-sel -/- versus ROSA26-LacZ+/E-sel +/+ mice in an ischemic wound model. Results: SDF-1α enhanced EPC-EC interaction by upregulating expression of E-sel/ligand in human EPC and HMVEC. Administration of SDF-1α in wound also increased E-sel expression on BMD- and circulating EPC in mice. The effects of SDF-1α on EPC-EC interaction and EPC transendothelial migration were specifically mediated by E-sel, as E-sel antagonists could inhibit these processes. E-sel was required for SDF-1α-induced EPC homing, wound neovascularization and healing in vivo (n=6, P<0.05). Conclusions: EPC-EC interaction is an essential process for EPC homing. SDF-1α induces both local luminal EC lining capillaries in wound tissue and remote BMD- and circulating EPC to express E-sel and its ligands. Up-regulated E-sel/ligand pairs reciprocally on EC and EPC serve as “Double-Lock” to secure targeted EPC-EC interactions selectively occurring within wound endothelium. These findings uncover a novel mechanism underlying the pro-angiogenic effect of SDF-1α on EPC homing and point to E-sel/ligand pairs reciprocally induced on circulating EPC and local endothelium as effective targets for ex vivo manipulation and therapeutic neovascularization

Abstract MP07: Nicotine Promotes Vascular Inflammation And Endothelial Dysfunction In Obese Rats

Epidemiological studies have shown that obesity and cigarette smoking (CS) are major cardiovascular (CV) risk factors and when coexisting in the same individuals have additive/synergistic effects upon CVD development. We have shown that stable compounds of CS (ATVB 2004) as well as nicotine, in concentrations found in smoker’s plasma, promote atherosclerotic CVD (AJP 2013). Here we studied in Sprague Dawley (SD) rats with diet -induced obesity the mechanisms involved in nicotine enhancement of CVD. SD rats (N=6-7 each group) were fed either a high fat (HF) or a standard chow (SCH) diet with or without nicotine (100 mg/kg/day in the drinking water) for 20 weeks. The HF rats developed central obesity, characterized by increased body weight gain (22%) and abdominal fat weight (53%), increased plasma levels of cholesterol (33%), non-esterified free fatty acids (68%), insulin (15%) glucose (12%)and systolic blood pressure (SBP: 146 ± 5 vs. 131 ± 5 mmHg SCH rats, p<0.05). Nicotine further increased SBP in obese HF rats (158 ± 4 mmHg, p<0.05) but not in lean SCH rats. Nicotine significantly increased O2- production in both obese (1689 ± 87 count/min/mg) and lean rats (1074 ± 105 count/min/mg) and further impaired EDR (Emax: 74 ± 5%) in obese HF rats. Nicotine also increased the expression of the macrophage marker ED1 in the aortas of obese HF rats. In peritoneal macrophages from obese HF rats TNFα, IL1β and CD36 were increased, and were further significantly increased in nicotine-treated obese HF rats. Using PCR array for inflammatory cytokines and receptor signaling pathway we found that the aortas from obese HF rats showed 2-4 fold increases in expression of several chemokines and interleukin genes expression; nicotine further increased the expression of 11 pro-inflammatory genes in the aortas from obese HF rats. Our results suggest that nicotine dramatically aggravates the CV effects of diet -induced obesity by increasing oxidative stress, vascular inflammation and endothelial dysfunction. Clinically chronic inhalation of nicotine, as that delivered by E-cigarettes, by individuals unable to quit the habit may have an important pro-atherogenic effect, particularly in obese subjects

Anti-inflammatory Properties of the Alpha-Melanocyte-Stimulating Hormone in Models of Granulomatous Inflammation

PURPOSE: Alpha-melanocyte stimulating hormone (α-MSH) is known to have anti-inflammatory effects. However, the anti-inflammatory properties of α-MSH on normal bronchial epithelial cells are largely unknown, especially in the context of in vitro sarcoidosis models. METHODS: We evaluated the anti-inflammatory effects of α-MSH on two different in vitro sarcoidosis models (lung-on-membrane model; LOMM and three-dimensional biochip pulmonary sarcoidosis model; 3D-BSGM) generated from NBECs and an in vivo sarcoidosis mouse model. RESULTS: Treatment with α-MSH decreased inflammatory cytokine levels and downregulated type I interferon pathway genes and related proteins in LOMM and 3D-BSGM models. Treatment with α-MSH also significantly decreased macrophages and cytotoxic T-cells counts in a sarcoidosis mice model. CONCLUSION: Our results confirm the direct role of type I IFNs in the pathogenesis of sarcoid lung granulomas and highlight α-MSH as a potential novel therapeutic agent for treating pulmonary sarcoidosis. GRAPHICAL ABSTRACT: [Image: see text] SUPPLEMENTARY INFORMATION: The online version contains supplementary material available at 10.1007/s00408-022-00546-x

Abstract 506: Nanocarrier-Targeted Mesenchymal Stem Cell for Angiogenesis and Wound Healing

Objectives: Wound healing and angiogenesis are impaired in patients with diabetes and peripheral vascular disease. We developed a novel treatment by systemic administration of mesenchymal stem cells (MSC) coated with nanocarrier composed of adhesion molecule -nanoparticle complex (AMNC). AMNC-MSC significantly enhanced wound healing and angiogenesis, in wild type and diabetic mice. AMNC-MSC targeted homing is specific to the injured tissue and superior to BSA (control)-NC coating or uncoated MSC. We now investigate the biosafety profile of the AMNC-MSC. Method: 1x 10 6 HUVEC were coated with AMNC. Cytotoxicity of AMNC was evaluated by trypan blue. Cell apoptosis was analyzed by flow cytometry using FITC-Annexin-V and propidium iodide. 12-14 week-old C57BL6 mice (N=15) underwent 6mm full thickness dorsal dermal wounding and then assigned to 3 IV treatment groups: (i) 1x 10 6 AMNC-MSC, (ii) 1x 10 6 MSC and (iii) equivalent volume saline (No Treatment (NT); (n=5/group). Day 8 post-wounding and treatment, comprehensive metabolic panel, complete blood count and urinalysis were obtained. Result: AMNC-MSC had no cytotoxicity to HUVEC. Blood tests and urinalysis revealed that the hematological, hepatic, renal and pancreatic functions were normal in all groups. Hyperglycemia was observed among all groups at comparable level (p=0.136), likely a stress response. All other blood/urine parameters were within normal, in all groups. Conclusion: AMNC-MSC has no observed cytotoxicity to human endothelial cells, in vitro . Intravenous administration of AMNC-MSC does not cause metabolic or hematologic toxicity, in mice. We demonstrate initial biosafety of novel nanocarrier-targeted MSC therapy which holds promise in pro-angiogenic and pro-healing regenerative medicine