Electrophilic PPARγ Ligands Attenuate IL-1β and Silica-Induced Inflammatory Mediator Production in Human Lung Fibroblasts via a PPARγ-Independent Mechanism

Acute and chronic lung inflammation is associated with numerous important disease pathologies including asthma, chronic obstructive pulmonary disease and silicosis. Lung fibroblasts are a novel and important target of anti-inflammatory therapy, as they orchestrate, respond to, and amplify inflammatory cascades and are the key cell in the pathogenesis of lung fibrosis. Peroxisome proliferator-activated receptor gamma (PPARγ) ligands are small molecules that induce anti-inflammatory responses in a variety of tissues. Here, we report for the first time that PPARγ ligands have potent anti-inflammatory effects on human lung fibroblasts. 2-cyano-3, 12-dioxoolean-1, 9-dien-28-oic acid (CDDO) and 15-deoxy-Δ12,14-prostaglandin J2 (15d-PGJ2) inhibit production of the inflammatory mediators interleukin-6 (IL-6), monocyte chemoattractant protein-1 (MCP-1), COX-2, and prostaglandin (PG)E2 in primary human lung fibroblasts stimulated with either IL-1β or silica. The anti-inflammatory properties of these molecules are not blocked by the PPARγ antagonist GW9662 and thus are largely PPARγ independent. However, they are dependent on the presence of an electrophilic carbon. CDDO and 15d-PGJ2, but not rosiglitazone, inhibited NF-κB activity. These results demonstrate that CDDO and 15d-PGJ2 are potent attenuators of proinflammatory responses in lung fibroblasts and suggest that these molecules should be explored as the basis for novel, targeted anti-inflammatory therapies in the lung and other organs

A Novel Anti-Inflammatory and Pro-Resolving Role for Resolvin D1 in Acute Cigarette Smoke-Induced Lung Inflammation

Introduction: Cigarette smoke is a profound pro-inflammatory stimulus that contributes to acute lung injuries and to chronic lung disease including COPD (emphysema and chronic bronchitis). Until recently, it was assumed that resolution of inflammation was a passive process that occurred once the inflammatory stimulus was removed. It is now recognized that resolution of inflammation is a bioactive process, mediated by specialized lipid mediators, and that normal homeostasis is maintained by a balance between pro-inflammatory and pro-resolving pathways. These novel small lipid mediators, including the resolvins, protectins and maresins, are bioactive products mainly derived from dietary omega-3 and omega-6 polyunsaturated fatty acids (PUFA). We hypothesize that resolvin D1 (RvD1) has potent anti-inflammatory and pro-resolving effects in a model of cigarette smoke-induced lung inflammation. Methods: Primary human lung fibroblasts, small airway epithelial cells and blood monocytes were treated with IL-1β or cigarette smoke extract in combination with RvD1 in vitro, production of pro-inflammatory mediators was measured. Mice were exposed to dilute mainstream cigarette smoke and treated with RvD1 either concurrently with smoke or after smoking cessation. The effects on lung inflammation and lung macrophage populations were assessed. Results: RvD1 suppressed production of pro-inflammatory mediators by primary human cells in a dose-dependent manner. Treatment of mice with RvD1 concurrently with cigarette smoke exposure significantly reduced neutrophilic lung inflammation and production of pro-inflammatory cytokines, while upregulating the anti-inflammatory cytokine IL-10. RvD1 promoted differentiation of alternatively activated (M2) macrophages and neutrophil efferocytosis. RvD1 also accelerated the resolution of lung inflammation when given after the final smoke exposure. Conclusions: RvD1 has potent anti-inflammatory and pro-resolving effects in cells and mice exposed to cigarette smoke. Resolvins have strong potential as a novel therapeutic approach to resolve lung injury caused by smoke and pulmonary toxicants

Expanding Co-Curricular Experiences: Global and Domestic Service Projects for Pharmacy Students

Presented at the FIP World Congress of Pharmacy and Pharmaceutical Sciences annual meeting in Buenos Aires, Argentina, in August 2016

Expanding Co-Curricular Experiences: Global and Domestic Service Projects for Pharmacy Students

Presented at the FIP World Congress of Pharmacy and Pharmaceutical Sciences annual meeting in Buenos Aires, Argentina, in August 2016

WSOP Prescription for Innovation—Use as Needed (PRN)

Objectives: The objective of this collaborative project is to showcase the educational innovations being conducted at the Wegmans School of Pharmacy. Methods: During the 2012-2013 school year, the Wegmans School of Pharmacy designed, implemented, and assessed a number of innovations in the areas of 1) didactic curriculum, 2) service learning, and 3) assessment. Examples of innovations include incorporation of service learning activities into didactic courses and experiential rotations, completion of a clinical trial, use of electronic rubrics, using pop culture to illustrate course objectives, peer teaching, blended learning, online flashcards, electronic review modules, electronic testing, and updates to classic active learning strategies like Turning Point, think/pair/share, and muddiest points. These innovations were assessed in a variety of ways, including the use of surveys, reflections, personal feedback, and exam scores. Results: Quantitative and qualitative analyses of educational innovations have revealed benefits to student learning, leadership ability, and experiences. Specifically, these innovations have been linked to improvements in student scores in courses and on national exams. They have also allowed for real-time improvements in faculty teaching. Overall, a majority of the innovations designed were well-received by students, and were seen as a positive addition to the educational experience. Implications: The continued improvement of pharmacy education requires response to the needs of learners, and innovations that push the achievements of our students. The innovations implemented in the 2012-2013 school year at Wegmans School of Pharmacy have been successful and beneficial to student learning and will be continued and expanded in future years

RvD1 inhibits IL-1β and CSE-induced cytokine production from primary human small airway epithelial cells and monocytes.

<p>Primary SAEC (<b>A–B</b>) were pre-treated with indicated concentrations of RvD1 for 24 hours then stimulated with IL-1β (1 ng/ml) for an additional 24 hours. Culture media was collected and levels of IL-6 (<b>A</b>) and IL-8 (<b>B</b>) were determined by ELISA. Data shown are mean ± SD of triplicate cultures, from one representative experiment of 3 performed. Blood-derived monocytes (<b>C–F</b>) were pre-treated with indicated concentrations of RvD1 for 24 hours then stimulated with IL-1β (1 ng/ml) or CSE (5%) for an additional 24 hours. Culture media was collected and levels of IL-6 (<b>C, E</b>) and IL-8 (<b>D, F</b>) were determined by ELISA. Data shown are mean ± SEM for three individual donors. ^##p<0.01, ^###p<0.001 compared to vehicle-treated control cultures. **p<0.01, ***p<0.001 compared to IL-1β or CSE without RvD1, using one-way ANOVA with Bonferroni post-tests.</p

17R-RvD1 promotes the resolution of acute lung inflammation induced by CS exposure.

<p>Mice were exposed to CS exposure on days 1, 2 and 3. The mice received 100 ng 17R-RvD1 (grey bars) or saline vehicle (Veh, black bars) by inhalation on day 3 (1 hour after the last cigarette smoke exposure), day 4 and day 5. Mice were euthanized at days 4 and 6 and differential cell counts were performed on BAL cells. Total cell number (<b>A</b>), number of neutrophils (<b>B</b>), and macrophages (<b>C</b>), percentage of neutrophils (D) and macrophages (E) are reported. BALF was collected and analyzed by ELISA or EIA for IL-6 (<b>F</b>), KC (<b>G</b>), MIP-2 (<b>H</b>) and PGE₂ (<b>I</b>). Data are shown as mean ± SEM for n  = 4–5 mice per group. N.D., not detected. *P<0.05, **P<0.01 compared to Vehicle/Smoke mice by two-way ANOVA with Bonferroni post-tests.</p

RvD1 drives polarization of alternatively activated macrophages.

<p>Mice were exposed to air or CS with or without RvD1 as described. (<b>A</b>) Expression of Arg-1 was measured in whole lung homogenates by Western blotting. Each lane represents one animal. <b>B–F:</b> Alveolar macrophages were enriched from BAL cells by adherence and total RNA was extracted and subjected to quantitative RT-PCR to measure mRNA levels of <i>Arg1</i> (<b>B</b>), <i>Mrc1</i> (<b>C</b>), <i>IL-10</i> (<b>D</b>), <i>iNOS</i> (<b>E</b>) and <i>TNFα</i> (<b>F</b>). Mean ± SEM is shown, normalized to 18S rRNA. Each group n = 6 for Arg1, Mrc1 and IL-10; n = 3 for iNOS and TNFα. ^#P<0.05, ^##P<0.01, compared to Vehicle/Air animal by one-way ANOVA with Bonferroni post-tests; *P<0.05, **P<0.01, ***P<0.001 versus Vehicle/Smoke animal by two-way ANOVA with Bonferroni post-test. ‡P<0.05 between RvD1/Air and RvD1/Smoke, ANOVA.</p

RvD1 decreases production of pro-inflammatory mediators in BALF and total lung homogenate but increases IL-10.

<p>Total protein (<b>A</b>) in BALF was measured by BCA assay. Neutrophil chemoattractants KC (<b>B</b>) and MIP-2 (<b>C</b>), IL-1β (<b>D</b>), IL-6 (<b>E</b>), MCP-1 (<b>F</b>), IFN-γ (<b>G</b>), PGE₂ (<b>H</b>) and IL-10 (<b>I</b>) were determined in lung homogenates by multiplex assay or EIA and normalized to total protein concentration. N.D., not detected. Data are shown as mean ± SEM for n = 6–8 mice per group and are representative of 3 independent experiments. *P<0.05, **P<0.01, ***P<0.001 versus smoke-exposed mice by two-way ANOVA with Bonferroni post-tests. ^###P<0.001 compared to air-exposed control mice by one-way ANOVA with Bonferroni post-tests. Expression of COX-2 and total actin (<b>J</b>) were determined in whole lung homogenates by Western blotting using COX-2 and total actin specific antibodies. Each lane represents an individual mouse.</p