Hyperplasia In Mice Research Articles

Glucagon-Like Peptide-1 Receptor Agonists and Thyroid Disorders

GLP-1 hormone secreted by L cells of intestine increases the glucose mediated insulin release, delay the gastric emptying and increases satiety by central mechanisms. Thus, a potential candidate for treatment of type 2 diabetes mellitus. Various GLP-1receptor analogues with longer half-life were introduced for the benefit of type 2 diabetes management. But animal studies of liraglutide showed increased incidence of thyroid cancers especially C cell hyperplasia in mice. This led to the safety concern of GLP-1RAs in terms of thyroid cancers. But further investigation showed that risk of thyroid cancers in humans is not significant as the receptor mapping in in vitro studies showed that in thyroid glands of humans and primates GLP-1 receptors were not expressed. Considering the safety, the drug was labelled as a contraindication for patients with MEN 2 syndrome or family history of Medullary thyroid cancer. Data from phase 3 trials of all the GLP-1RAs did not show any significant increase in thyroid cancer or any cancer with GLP-1 usage. Meta-analysis of the trials also showed no significant increase in thyroid or any other cancer in comparison to comparators. But two observational studies showed the increase in incidence of thyroid cancers among patients on GLP-RAs, but the studies showed limitations like study design limitation, unaccounted biases, over reporting etc. Surveillance data and review of all available data shows that there is no significant increase in risk of any thyroid disorder and treatment decision to be on risk benefit basis.

PD53-08 E-CIGARETTE EXPOSURE IN MICE PROMOTES CHANGES IN DNA METHYLATION OF THE BLADDER UROTHELIUM

You have accessJournal of UrologyCME1 May 2022PD53-08 E-CIGARETTE EXPOSURE IN MICE PROMOTES CHANGES IN DNA METHYLATION OF THE BLADDER UROTHELIUM Ibardo Zambrano, Ujjawal Manocha, Weida Gong, Yi-Hsuan Tsai, Manhua Zhu, Melissa Herman, Marc Bjurlin, and William Kim Ibardo ZambranoIbardo Zambrano More articles by this author , Ujjawal ManochaUjjawal Manocha More articles by this author , Weida GongWeida Gong More articles by this author , Yi-Hsuan TsaiYi-Hsuan Tsai More articles by this author , Manhua ZhuManhua Zhu More articles by this author , Melissa HermanMelissa Herman More articles by this author , Marc BjurlinMarc Bjurlin More articles by this author , and William KimWilliam Kim More articles by this author View All Author Informationhttps://doi.org/10.1097/JU.0000000000002630.08AboutPDF ToolsAdd to favoritesDownload CitationsTrack CitationsPermissionsReprints ShareFacebookLinked InTwitterEmail Abstract INTRODUCTION AND OBJECTIVE: To date, no group has studied the epigenetic effects of e-cigarettes (EC) on the urothelium. DNA methylation changes appear to be one of the earliest events in bladder tumorigenesis and can be readily found in normal urothelium adjacent to bladder tumors. EC exposure causes urothelial hyperplasia in mice but not bladder tumors. We Vtherefore asked whether EC exposure results in early field alterations of DNA methylation in the bladder urothelium. METHODS: Mice (4 females, 4 males; 35 weeks old) were exposed to 12% nicotine in 1:1 polyethylene glycol: vegetable glycerol or vehicle daily: 3 hr sessions, 3 sec vape duration, 10 min interval between vapes. Plasma nicotine and cotinine serum concentrations were measured using LC-MS. Reduced-representation bisulfite sequencing (RRBS) was performed using DNA from microdissected urothelium harvested at 4 weeks (Zymo Research Kit). Number of loci covered by each sample was determined and data was filtered to only include loci with >10 reads. Differentially methylated loci were determined (p<0.01). Gene Set Enrichment Analysis was performed to contextualize the DNA methylation changes. RESULTS: No significant adverse events were observed in the mice during the exposure period to either vehicle or nicotine. Acute and repeated exposure resulted in significantly higher levels of nicotine and cotinine serum concentration in the nicotine vs. the vehicle group (Figure 1). No bladder tumors formed. RRBS analysis demonstrated that EC exposure resulted in hypermethylation of the promoters of 665 genes and hypomethylation of 285 genes after accounting for sex (p <0.05) Pathway analysis showed enrichment of MYC target genes in hypomethylated regions and enrichment of genes associated with oxidative phosphorylation, cilium assembly, and ciliopathies in hypermethylated loci. CONCLUSIONS: Significant changes of DNA methylation were observed in mice exposed to ECS vapor vs. vehicle control. EC metabolites in urine may result in aberrant methylation patterns in the bladder urothelium. Results are suggestive of a pre-malignant phenotype driven by EC exposure via stimulation of MYC targets and silencing of genes responsible for establishing primary cilia. Loss of primary cilia in epithelial cells has been associated with loss of cell polarity and increased cell migration. Source of Funding: NCI Geographical Management of Cancer Health Disparities Program (3P30-CA016086), UNC Lineberger Innovation Award 2020 © 2022 by American Urological Association Education and Research, Inc.FiguresReferencesRelatedDetails Volume 207Issue Supplement 5May 2022Page: e912 Advertisement Copyright & Permissions© 2022 by American Urological Association Education and Research, Inc.MetricsAuthor Information Ibardo Zambrano More articles by this author Ujjawal Manocha More articles by this author Weida Gong More articles by this author Yi-Hsuan Tsai More articles by this author Manhua Zhu More articles by this author Melissa Herman More articles by this author Marc Bjurlin More articles by this author William Kim More articles by this author Expand All Advertisement PDF DownloadLoading ...

Trichoderma longibrachiatum-Associated Skin Inflammation and Atypical Hyperplasia in Mouse.

BackgroundThe relationship between infection and tumors has attracted increasing attention. Trichoderma spp. are often isolated from tumors. However, their potential role remains unclear. We recently reported the isolation of Trichoderma longibrachiatum from a patient with pulmonary spindle cell carcinoma that was confirmed as primary infection by application of laser capture microdissection and polymerase chain reaction. To explore whether the strain is pathogenic and whether it can cause atypical cell proliferation and infiltration of NK cells and T cells, we designed a mouse infection experiment.MethodsTwelve ICR mice were randomly separated into three groups. Cyclophosphamide was used to inhibit the immunity of mice. A mouse model of Trichoderma infection was successfully established by intracutaneous injection on the back skin with a suspension of strain PKUT180420015. The pathological manifestations of Trichoderma infection and the interaction between immune cells and fungi were observed by histopathology, immunohistochemistry and intensive fungal staining. Reisolation of the fungus was observed by infected tissue culture. The inoculated sites exhibited swelling 3 days after inoculation, and ulcers developed at approximately 14 days. Skin specimens were obtained and then cultured at 3, 7, and 14 days after inoculation. We selected mice 14 days after inoculation in Group 3, whose ulcers were the most typical, for histological analysis.ResultsInflammation, angioinvasion and necrosis were observed. Immunohistochemistry showed positive markers of Ki67, CD3, CD56, GZMB, and PRF. Periodic acid-Schiff staining, periodic acid-silver methenamine staining, and Calcofluor staining showed fungal spores in the vascular lumen, vascular walls and around the blood vessels.ConclusionsOur studies showed that a T. longibrachiatum strain (PKUT180420015) isolated from a biopsy specimen in a patient with pulmonary spindle cell carcinoma could induce atypical hyperplasia, with the expression of Ki67, CD3, CD56, GZMB, and PRF in mice. These data indicate that the fungus may be involved in inducing atypical hyperplasia or tumorigenesis.

Histopathological and reproductive effect of tamsulosin and finasteride on induced Benign prostate hyperplasia in mice

The prostate hyperplasia (BPH) was induced in male mice by subcutaneous injection of testosterone propionate (20 mg/kg) for 30 days. Fifty mice were divided equally into five groups, the first group (G1) was (negative control group), and the second group (G2) was (positive Control group) induced BPH previously and inoculated orally distilled water. The third group (G3) was induced BPH and treated orally with tamsulosin (0.156mg /kg), the fourth group (G4) was induced BPH and treated orally with Finasteride (1.495 mg /kg) and finally, the fifth (G5) was induced BPH and treated orally with a combination of tamsulosin and finasteride (0.0825, 0.7475) mg/kg respectively. The histopathological results showed prostate hyperplasia and abnormal proliferation of the epithelial and stromal cells with inflammation cells accompanying stenosis of acini in G2. Whereas showed a decrease in hyperplasia, and epithelial layer thickness in G3 and G4. Furthermore, prostate shrinkage and the glandular cavities trended towards the normal approach of the prostate in G5. The testosterone levels in G2 showed a significant increase (P&lt;0.05) as compared to other treated groups with a more significant decrease in G4 compared to other treated groups.

Ethanol Extract of Chaenomeles sinensis Inhibits the Development of Benign Prostatic Hyperplasia by Exhibiting Anti-oxidant and Anti-inflammatory Effects.

Chaenomeles sinensis is known to inhibit the development and progression of many age-related diseases, but the underlying molecular mechanisms are largely unclear. In the present study, we observed that the ethanol extract of Chaenomeles sinensis scavenged 2,2’-diphenylpicrylhydrazyl and 2,2’-azinobis diammonium radicals in vitro. The ethanol extract of Chaenomeles sinensis activated antioxidant response element-luciferase activity and induced expression of NRF2 target genes in HaCaT cells. The ethanol extract of Chaenomeles sinensis also suppressed LPS-induced expression of COX-2 and iNOS proteins, and mRNA expression of TNF-α and IL-2 in RAW264.7 cells. Finally, the ethanol extract of Chaenomeles sinensis significantly suppressed testosterone propionate-induced benign prostatic hyperplasia in mice. Together, our study provides the evidence that the ethanol extract of Chaenomeles sinensis inhibits the development of benign prostatic hyperplasia by exhibiting anti-oxidant and anti-inflammatory effects.

A murine MODEL of e-cigarettes exposure to evaluate the carcinogenic effects of vaping on the bladder urothelium.

566 Background: Data evaluating the carcinogenic potential of e-cigarette (ECS) by-products on the bladder urothelium is scarce, and to date, no group has studied the direct impact of ECS on the urothelium at the genomic and epigenetic level. While it has been published that ECS exposure causes bladder hyperplasia in mice, no bladder tumors formed and the field is therefore lacking appropriate preclinical models to be able to study ECS related bladder cancer. Importantly, whole organ mapping studies of tumor bearing human bladders have identified DNA methylation field changes in “adjacent normal” urothelium as early events in bladder carcinogenesis. Whether ECS exposure results in early field alterations of DNA methylation in the urothelium is unknown. Methods: Mice (4 females, 4 males; 35 weeks old) were exposed to 12% nicotine (120mg/ml) in 1:1 polyethylene glycol: vegetable glycerol (PG/VG) or vehicle (PG/VG) daily: 3 hr sessions, 3 sec vape duration, 10 min interval between vapes. Nicotine and cotinine serum concentrations were measured at 3 hrs, and then at 5 days using LC-MS. Bladders were harvested after 4 weeks. DNA was extracted from microdissected urothelial layer. Reduced representation bisulfite sequencing (RRBS) was performed (Zymo Research) using 100ng of DNA input and paired-end chemistry. Alignment was performed using Bismark. Number of loci covered by each sample was determined and data was filtered to only include loci with &gt; 10 reads. Differentially methylated loci were determined. Adjusted p-value &lt; 0.01 was used as threshold to generate heatmaps. Gene Set Enrichment Analysis was performed to contextualize the DNA methylation changes. Results: Acute and repeated exposure resulted in significantly higher levels of nicotine and cotinine serum concentration in the nicotine group vs. the vehicle group without significant adverse events. No tumors were observed on gross visualization at the time of urothelium harvest. The number of loci in the RRBS libraries with &gt; 10 reads ranged from 130,000 to 250,000. 53 loci were differentially methylated between nicotine and vehicle. These loci did not correspond to the top hyper/hypo methylated genes in premalignant bladder tissue from recently reported whole-cystectomy mapping. Conclusions: We established an effective and safe murine model of ECS exposure closely resembling the human experience of vaping. Significant changes of DNA methylation were observed in mice exposed to ECS vapor vs. a vehicle control after four weeks of daily exposure. Ongoing experiments include longer vaping exposures as well as comparison of RRBS to a recently developed commercial mouse methylarray. Understanding epigenetic and genomic changes as a result of ECS exposure in preclinical models can serve as a foundation for studies developing assays to identify ECS users whose bladders contain early field defects that may place them at higher risk for bladder cancer.

Deletion of Smooth Muscle Lethal Giant Larvae 1 Promotes Neointimal Hyperplasia in Mice.

Vascular smooth muscle cell (VSMC) proliferation and migration contribute to neointimal hyperplasia after injury, which causes vascular remodeling related to arteriosclerosis, hypertension, and restenosis. Lethal giant larvae 1 (LGL1) is a highly conserved protein and plays an important role in cell polarity and tumor suppression. However, whether LGL1 affects neointimal hyperplasia is still unknown. In this study, we used smooth muscle-specific LGL1 knockout (LGL1SMKO) mice generated by cross-breeding LGL1flox/flox mice with α-SMA-Cre mice. LGL1 expression was significantly decreased during both carotid artery ligation in vivo and PDGF-BB stimulation in vitro. LGL1 overexpression inhibited the proliferation and migration of VSMCs. Mechanistically, LGL1 could bind with signal transducer and activator of transcription 3 (STAT3) and promote its degradation via the proteasomal pathway. In the carotid artery ligation animal model, smooth muscle-specific deletion of LGL1 accelerated neointimal hyperplasia, which was attenuated by the STAT3 inhibitor SH-4-54. In conclusion, LGL1 may inhibit neointimal hyperplasia by repressing VSMC proliferation and migration via promoting STAT3 proteasomal degradation.

Effects of Wood Smoke Constituents on Mucin Gene Expression in Mice and Human Airway Epithelial Cells and on Nasal Epithelia of Subjects with a Susceptibility Gene Variant in Tp53.

Background:Exposure to wood smoke (WS) increases the risk for chronic bronchitis more than exposure to cigarette smoke (CS), but the underlying mechanisms are unclear.Objective:The effect of WS and CS on mucous cell hyperplasia in mice and in human primary airway epithelial cells (AECs) was compared with replicate the findings in human cohorts. Responsible WS constituents were identified to better delineate the pathway involved, and the role of a tumor protein p53 (Tp53) gene polymorphism was investigated.Methods:Mice and primary human AECs were exposed to WS or CS and the signaling receptor and pathway were identified using short hairpin structures, small molecule inhibitors, and Western analyses. Mass spectrometric analysis was used to identify active WS constituents. The role of a gene variant in Tp53 that modifies proline to arginine was examined using nasal brushings from study participants in the Lovelace Smokers Cohort, primary human AECs, and mice with a modified Tp53 gene.Results:WS at 25-fold lower concentration than CS increased mucin expression more efficiently in mice and in human AECs in a p53 pathway-dependent manner. Study participants who were homozygous for p53 arginine compared with the proline variant showed higher mucin 5AC (MUC5AC) mRNA levels in nasal brushings if they reported WS exposure. The WS constituent, oxalate, increased MUC5AC levels similar to the whole WS extract, especially in primary human AECs homozygous for p53 arginine, and in mice with a modified Tp53 gene. Further, the anion exchange protein, SLC26A9, when reduced, enhanced WS- and oxalate-induced mucin expression.Discussion:The potency of WS compared with CS in inducing mucin expression may explain the increased risk for chronic bronchitis in participants exposed to WS. Identification of the responsible compounds could help estimate the risk of pollutants in causing chronic bronchitis in susceptible individuals and provide strategies to improve management of lung diseases. https://doi.org/10.1289/EHP9446

Guifu Dihuang Pills Ameliorated Mucus Hypersecretion by Suppressing Muc5ac Expression and Inactivating the ERK-SP1 Pathway in Lipopolysaccharide/Cigarette Smoke-Induced Mice.

Mucus hypersecretion is a hallmark of chronic obstructive pulmonary disease (COPD) and is associated with increasing sputum production and declining pulmonary function. Therefore, reducing mucus secretion can be a new therapeutic opportunity for preventing COPD. The Guifu Dihuang pill (GFDHP) is a classical Chinese medicine and has been used as an immunoregulator for treatment of kidney yang deficiency syndrome, including hypothyroidism, adrenocortical hypofunction, chronic bronchitis, and COPD, for more than 2000 years. However, the protective effects and mechanisms of GFDHP against mucus hypersecretion in COPD remain obscure. The aim of the present study was to explore the inhibitory effects of GFDHP on lipopolysaccharide/cigarette smoke- (LPS/CS-) induced Mucin5ac (Muc5ac) overproduction and airway goblet cell hyperplasia in mice. The mice were randomly assigned into 6 groups: control, model, GFDHP-L, GFDHP-M, GFDHP-H, and dexamethasone. The mice were given LPS twice through intranasal inhalation and then exposed to CS daily for 6 weeks. Three doses of GFDHP were orally administered daily during the last 3 weeks of the experiment. Pulmonary function was examined with an EMKA pulmonary system, and pulmonary hyperpermeability and lung damage were evaluated with an in vivo imaging system. Inflammatory cells and cytokines in bronchoalveolar lavage fluid (BALF) were detected with a cell count analyzer and though ELISA analysis, respectively. Lung pathological changes and airway goblet cell hyperplasia were analyzed with hematoxylin and eosin and Alcian blue periodic acid Schiff staining. The protein expression levels of Muc5ac and extracellular signal-regulated kinase (ERK)-specificity protein1 (SP1) signaling pathway were measured with Western blot and immunohistochemistry. The results demonstrated that GFDHP improved pulmonary function and suppressed mouse pulmonary hyperpermeability and edema. GFDHP suppressed inflammatory cell infiltration and cytokine release in BALF, thereby elevating pulmonary function. It ameliorated lung pathological changes and airway goblet cell hyperplasia, and suppressed expression levels of Muc5ac mRNA and protein and phospho-ERK and SP1 levels in the lung tissues of the COPD mice. In conclusion, GFDHP inhibited mucus hypersecretion induced by LPS/CS by suppressing the activation of the ERK-SP1 pathway.

Экспериментальное исследование эффективности ректальных свечей с окисленным декстраном при гиперплазии предстательной железы

AIM to evaluate therapeutic activity of oxidized dextran after rectal administration in a modified sulpiride model of benign prostatic hyperplasia in mice. MATERIALS AND METHODS The therapeutic activity of oxidized dextran was studied in an experimental model of benign prostatic hyperplasia in mice (sulpiride model). All in vivo studies were carried out on 30 male BALB/c mice with an average body weight of 30-35 g. RESULTS It was found that rectal administration of a solution of oxidized dextran effectively neutralizes pathomorphological changes in experimental model of benign prostatic hyperplasia. In the group of animals, which received 2% solution of oxidized dextran as treatment, the most prominent therapeutic effect in an experimental model was seen and it was confirmed by a decrease in the bladder detrusor thickness and the normalization of its lumen.

Aerobic exercises ameliorate benign prostatic hyperplasia via IGF-1/IGF-1R/ERK/AKT signalling pathway in prostate tissue of high-fat-diet-fed mice with insulin resistance

This study investigated the changes in the prostate of high-fat diet (HFD)-fed mice with insulin resistance (IR) and explored the possible mechanisms of the effects of 8-week treadmill aerobic exercise on prostatic hyperplasia in insulin-resistant mice through the IGF-1/IGF-1R/ERK/AKT signalling pathway. Results showed IR in mice caused an increase in prostate-related indicators, such as prostate weight (PW) and prostate volume (PV), resulting in prostatic hyperplasia. The area of the glandular lumen and the height of the glandular epithelium in mice with IR were increased, which indicating that it caused prostatic hyperplasia through epithelial cell proliferation. In addition, the level of IGF-1 in serum and the expression of IGF-1R, ERK and AKT in prostate tissue of high-fat diet induced IR mice increased significantly, which might be related to the proliferation of prostate cells. However, aerobic exercise lowered the blood sugar, serum insulin and IGF-1; inhibited the combination of IGF-1 and IGF-1R on the prostate; down-regulated the expression of IGF-1R, ERK and AKT proteins; and then suppressed the expression of downstream proliferation genes, thereby achieving the purpose of inhibiting the proliferation of prostate epithelial cells. In conclusion. Eight weeks of aerobic exercise might improve the prostate hyperplasia in mice via down-regulating the serum insulin and IGF-1, thus enhancing the insulin sensitivity of insulin-resistant mice and regulating the IGF-1/IGF-1R/ERK/AKT signalling pathway by inhibiting the expression of IGF-1R, ERK and AKT in the prostate tissue. However, this exercise had no significant effect on PV, PW and prostate index (PI).

Treatment with broad-spectrum antibiotics upregulates Sglt1 and induces small intestinal villous hyperplasia in mice.

Although extensive evidence indicates that the gut microbiota plays a crucial role in regulating glucose homeostasis, the exact regulatory mechanism remains unclear. This study aimed to investigate the effect of broad-spectrum antibiotics on the expression of glucose transporters, histomorphology of the small intestine, and glucose metabolism in mice. C57BL/6 mice were administered drinking water with or without a broad-spectrum antibiotic combination for 4 weeks. Thereafter, an oral glucose tolerance test was performed. Body weight, small intestine histopathology, mRNA levels of glucose transporters (SGLT1 and GLUT2) and intestinal transcription factors (CDX1 and CDX2) were evaluated. SGLT1 and CDX1 were upregulated in the small intestine upon antibiotic administration compared with that in the control group. The height and surface area of the jejunal villi were significantly higher upon antibiotic administration than in the control group. Fasting glucose levels were significantly higher upon antibiotic administration than in the control group. The present results indicate that treatment with broad-spectrum antibiotics upregulates SGLT1 and CDX1 and induces hyperplasia in the small intestine, thus increasing fasting blood glucose levels. Our results further the current understanding of the effects of broad-spectrum antibiotics on the gut microbiota and glucose homeostasis that may have future clinical implications.

Guizhi Fuling Capsule ameliorates endometrial hyperplasia through promoting p62-Keap1-NRF2-mediated ferroptosis

Guizhi Fuling Capsule (GFC) is a classical traditional Chinese medicine officially recorded in Synopsis of the Golden Chamber and has long been used to treat gynecological diseases in China. However, scientific evidence for the anti-endometrial hyperplasia potential of GFC used in traditional medicine is lacking. This study evaluated whether GFC protects against endometrial hyperplasia and its potential mechanism in mice. We used estrogen (estradiol) to induce endometrial hyperplasia in mice. C57BL/6 mice were treated with estradiol subcutaneously for 21 days, and GFC (75mg/kg and 150mg/kg) was given intragastric administration from the first day of the modeling. H&E staining is used to evaluate endometrial tissue structure change. Malondialdehyde was measured to explore lipid peroxidation. Western blot, immunohistochemistry and immunofluorescence were performed to observe the expressions of GPX4, p62, Keap1 and NRF2. The degree of ferroptosis in endometrial tissue of patients with endometrial hyperplasia was lower than normal endometrial tissue. In addition, ferroptosis inducer imidazole ketone erastin could improve endometrial hyperplasia in mice. Interestingly, GFC significantly alleviated endometrial hyperplasia through triggering ferroptosis. Furthermore, GFC inhibited p62-Keap1-NRF2 pathway in estradiol-induced endometrial hyperplasia model. GFC may attenuate estrogen-induced endometrial hyperplasia in mice through triggering ferroptosis via inhibiting p62-Keap1-NRF2 pathway. These findings suggest that GFC might act as a promising traditional Chinese medicine to treat endometrial hyperplasia.

Smooth Muscle Cell-Specific PKM2 (Pyruvate Kinase Muscle 2) Promotes Smooth Muscle Cell Phenotypic Switching and Neointimal Hyperplasia.

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Bacillus thuringiensis Cry1Ac toxin and protoxin do not provoke acute or chronic cytotoxicity on macrophages and leukocytes.

The bioinsecticidal Cry1Ac proteins (protoxin and toxin) are potent immunogens that can activate macrophages by inducing upregulation of costimulatory molecules, pro-inflammatory cytokines, and mitogen-activated protein kinase (MAPK) signaling pathways. Besides, by the oral route, Cry1Ac toxin is mildly allergenic and induces intestinal lymphoid hyperplasia in mice. Given the potential utility of Cry1Ac protoxin as an adjuvant, as well as the human consumption of Cry1Ac toxin in transgenic crops, it is necessary to more deeply evaluate the toxicological potential of these proteins in mammalian immune cells. Here, were used in vitro evaluations in leukocyte and macrophage cell lines to test the potential toxicity of various doses of Cry1Ac proteins, by means of Alamar Blue, MTT, Annexin V, and JC1 assays. Our results indicated that neither Cry1Ac protoxin nor toxin elicited acute toxic effects, after monitoring the cell activity for 4, 8, 10, and 24h of exposure. By flow cytometry and confocal microscopy analysis, it was observed that neither Cry1Ac toxin nor protoxin generated mitochondrial damage or depolarization or induced apoptosis or necrosis. In conclusion, despite their immunostimulatory effects, it was demonstrated that Cry1Ac proteins did not have cytotoxic effects, even at high concentrations, in primary leukocytes or macrophages or cell lines.

Evaluating the Interactions between Endothelial and Smooth Muscle Cells in the Presence of Drug-Eluting Stents and Platelets: Is Interleukin6 a New Possible Therapeutic Target for In-Stent Restenosis?

The success of percutaneous coronary intervention (PCI) procedures is limited by in-stent restenosis (ISR). The use of drug-eluting stents (DES) decreased the incidence of ISR; however, ISR continues to affect a percentage of patients with ischemic cardiac events. Therefore, there is room for improvement considering not only the effectiveness of DES but also the adjunct pharmacological interventions for patients after PCI. Interleukin 6 (IL-6) can stimulate the growth of vascular smooth muscle cells (SMCs) in a platelet-derived growth factor (PDGF)-dependent manner and that antibodies against the IL-6 receptor reduced intimal hyperplasia in mice. In this study, our aim is to develop an in vitro study, based on co-cultures of endothelial cells (EC) and SMCs in the presence of platelets and stents, to analyze the cytokine responses (IL-6 and other inflammation markers) associated with ISR, in the quest for new therapeutic targets. Overall, in the presence of platelet-rich plasma (PRP), there was an increase in the production of reactive oxygen species in the groups that received the stents, probably because platelets potentiate stent-associated oxidative stress. Moreover, the analysis of the expression of inflammatory markers showed a difference in the expression of IL-6 only in the groups that received stents, which further increased in the presence of platelets. Altogether, our findings suggest that the down regulation of IL-6 expression using pharmacologic inhibitors is a possible new therapeutic strategy in the context of ISR.

TRPA1 deficiency alleviates inflammation of atopic dermatitis by reducing macrophage infiltration

AimsThe aim of this study was to investigate the role of TRPA1 in the pathogenesis of AD. Main methodsThe experimental atopic dermatitis (AD)-like skin lesions were established using 2,4-dinitrochlorobenzene (DNCB). Mice were divided into three groups: TRPA1−/− and WT groups were treated with DNCB dissolved in a 3:1 mixture of acetone and olive oil; the negative control group was treated with 3:1 mixture of acetone and olive oil without DNCB. The treatment lasted for 21 days, after which the animals were sacrificed and their blood, ears and dorsal skin tissue samples were collected for analysis. Key findingsLower dermatitis score, ear thickness, pruritus score, and epidermal hyperplasia were observed in mice in TRPA1−/− mice compared to the WT group. Besides, lower dermal mast cell infiltration, proinflammatory cytokines, Th2 cytokines and the infiltration of macrophages were observed in the TRPA1−/− mice compared to the WT group. Furthermore, we demonstrated that TRPA1 antagonist HC-030031 could alleviate AD-like symptoms and reduce the degree of epidermal hyperplasia in mice. SignificanceTRPA1 has a crucial role during the AD pathogenesis in mice, thus may be used as a potential new target for treating patients with chronic skin inflammatory disease.

Macrophage NCOR1 Deficiency Ameliorates Myocardial Infarction and Neointimal Hyperplasia in Mice.

BackgroundNCOR1 (nuclear receptor corepressor 1) is an essential coregulator of gene transcription. It has been shown that NCOR1 in macrophages plays important roles in metabolic regulation. However, the function of macrophage NCOR1 in response to myocardial infarction (MI) or vascular wire injury has not been elucidated.Methods and ResultsHere, using macrophage Ncor1 knockout mouse in combination with a mouse model of MI, we demonstrated that macrophage NCOR1 deficiency significantly reduced infarct size and improved cardiac function after MI. In addition, macrophage NCOR1 deficiency markedly inhibited neointimal hyperplasia and vascular remodeling in a mouse model of arterial wire injury. Inflammation and macrophage proliferation were substantially attenuated in hearts and arteries of macrophage Ncor1 knockout mice after MI and arterial wire injury, respectively. Cultured primary macrophages from macrophage Ncor1 knockout mice manifested lower expression of inflammatory genes upon stimulation by interleukin‐1β, interleukin‐6, or lipopolysaccharide, together with much less activation of inflammatory signaling cascades including signal transducer and activator of transcription 1 and nuclear factor‐κB. Furthermore, macrophage Ncor1 knockout macrophages were much less proliferative in culture, with inhibited cell cycle progression compared with control cells.ConclusionsCollectively, our data have demonstrated that NCOR1 is a critical regulator of macrophage inflammation and proliferation and that deficiency of NCOR1 in macrophages attenuates MI and neointimal hyperplasia. Therefore, macrophage NCOR1 may serve as a potential therapeutic target for MI and restenosis.

Lactobacillus reuteri maintains intestinal epithelial regeneration and repairs damaged intestinal mucosa

ABSTRACT Little is known about the regulatory effect of microbiota on the proliferation and regeneration of ISCs. Here, we found that L. reuteri stimulated the proliferation of intestinal epithelia by increasing the expression of R-spondins and thus activating the Wnt/β-catenin pathway. The proliferation-stimulating effect of Lactobacillus on repair is further enhanced under TNF -induced intestinal mucosal damage, and the number of Lgr5+ cells is maintained. Moreover, compared to the effects of C. rodentium on the induction of intestinal inflammation and crypt hyperplasia in mice, L. reuteri protected the intestinal mucosal barrier integrity by moderately modulating the Wnt/β-catenin signaling pathway to avoid overactivation. L. reuteri had the ability to maintain the number of Lgr5+ cells and stimulate intestinal epithelial proliferation to repair epithelial damage and reduce proinflammatory cytokine secretion in the intestine and the LPS concentration in serum. Moreover, activation of the Wnt/β-catenin pathway also induced differentiation toward Paneth cells and increased antimicrobial peptide expression to inhibit C. rodentium colonization. The protective effect of Lactobacillus against C. rodentium infection disappeared upon application of the Wnt antagonist Wnt-C59 in both mice and intestinal organoids. This study demonstrates that Lactobacillus is effective at maintaining intestinal epithelial regeneration and homeostasis as well as at repairing intestinal damage after pathological injury and is thus a promising alternative therapeutic method for intestinal inflammation.

Toll-like receptor 9 regulates metabolic profile and contributes to obesity-induced benign prostatic hyperplasia in mice.

Benign prostatic hyperplasia (BPH) is associated with obesity and prostatic inflammation. The present study investigated the participation of toll-like receptor 9 (TLR9) in obesity-induced BPH, focusing on metabolic impairments, damage-associated molecular patterns (DAMP) levels and prostatic oxidative stress generation. C57BL/6 (WT) and TLR9 mutant male mice were fed with regular or high-fat diet for 12 weeks. Metabolic profile, functional protocols, reactive-oxygen species (ROS) generation, prostatic histological analysis and DAMP levels were analyzed. Western blotting for prostatic TLR9 signaling pathway was also performed. BPH in WT obese animals was characterized by increased prostate weight, smooth muscle hypercontractility and prostatic epithelial hyperplasia. Higher epididymal fat weight and prostatic ROS generation along with increased fasting glucose, triglyceride and circulating DAMP levels were also observed in WT obese group. Conversely, TLR9 mutant obese animals exhibited lower epididymal fat weight, fasting glucose and triglyceride levels associated with reduced prostate hypercontractility, prostatic ROS and circulating DAMP levels. However, TLR9 mutant obese mice were not protected from obesity-associated prostatic overgrowth and epithelial hyperplasia. Interestingly, TLR9 mutant lean mice exhibited augmented fasting glucose and prostatic ROS levels compared with WT lean mice. Despite increased prostatic expression of TLR9 in WT obese mice, no differences were seen in MyD88 expression between groups. Improved obesity-induced BPH-related prostatic smooth muscle hypercontractility in TLR9 obese mice may be associated with amelioration in the metabolic profile, ROS and DAMP generation. Therefore, TLR9 could be a valuable target to improve obesity-associated metabolic disorders and prostate smooth muscle hypercontractility in BPH.