Downregulation Of Matrix Metalloproteinase Research Articles

Expression and potential role of CXCL5 in the pathogenesis of intrauterine adhesions.

ObjectiveC-X-C motif chemokine ligand 5 (CXCL5), a member of the chemokine family, is associated with remodeling of connective tissues. However, its role in formation of intrauterine adhesions (IUA) remains unclear. We aimed to investigate the expression and mechanism underlying the role of CXCL5 in IUA.MethodsExpression of CXCL5 in IUA was detected by immunohistochemistry in a rat model of IUA and by real-time PCR and western blotting in patients with IUA. The protein levels of matrix metalloproteinase 9 (MMP9) and transcription factor p65 in human endometrial cells were assessed by western blotting after CXCL5 overexpression.ResultsProtein expression of CXCL5 was significantly decreased in the endometria of IUA rats compared with that of control and sham-operated rats. Real-time PCR and western blotting in patients with IUA showed similar results to those from the rat model. After overexpression, CXCL5 significantly upregulated expression of MMP9 and slightly upregulated expression of p65 in human endometrial cells.ConclusionsCXCL5 plays an important role in IUA formation after endometrial injury. We propose a molecular mechanism to explain formation of IUA, including downregulation of MMP9 by low CXCL5 expression. These findings provide valuable information for the prevention and targeted therapy of IUA.

Berberine accelerated wound healing by restoring TrxR1/JNK in diabetes.

The high disability, mortality and morbidity of diabetic ulcers make it urgent to explore effective strategies for diabetic wound repair. TrxR1 plays a vital role in regulating redox homeostasis in various pathologies. In the present study, the effect of berberine (BBR) on diabetic wounds was investigated in streptozotocin (STZ)-induced diabetic rats and a high glucose (HG)-induced cell model, and the mechanism of BBR on TrxR1 was elucidated. BBR treatment remarkably accelerated wound healing and enhanced extracellular matrix (ECM) synthesis and significantly inhibited HG-induced HaCaT cell damage. Further analysis indicated that BBR activated TrxR1, suppressed its downstream JNK signaling, thereby inhibiting oxidative stress and apoptosis, promoted cell proliferation, down-regulated matrix metalloproteinase (MMP) 9 (MMP9) and up-regulated transforming growth factor-β1 (TGF-β1) and tissue inhibitors of MMP 1 (TIMP1), resulting in accelerated wound healing. Importantly, the enhancement of BBR on wound repair was further abolished by TrxR1 inhibitor. Moreover, in diabetic wounds induced by a combination of STZ injection and high-fat diet, BBR significantly increased wound closure rate and TrxR1 expression, and this was reversed by TrxR1 inhibitor. These data indicated that topical BBR treatment accelerated diabetic wound healing by activating TrxR1. Targeting TrxR1 may be a novel, effective strategy for restoring redox homeostasis and promoting diabetic wound healing.

Trichostatin A and Zebularine along with E-cadherin re-expression enhance tumour necrosis factor-related apoptosis-inducing ligand (TRAIL)-mediated cell cycle arrest in human breast adenocarcinoma cells

Breast cancer is the leading cause of death among women in which its definite cure remains uncovered. Tumour necrosis factor-related apoptosis inducing ligand (TRAIL) is a potential anti-cancer agent due to its selective killing towards cancer cells while sparing the healthy cells. However, it is limited by the development of TRAIL resistance. With the attempt to overcome TRAIL resistance, this research embarked to study the effect of epigenetic drugs, Trichostatin A (TSA) and Zebularine (Zeb) along with E-cadherin re-expression on anti-cancer effect in human breast adenocarcinoma cells. The MDA-MB-231 re-expressed with E-cadherin (231-EGFP) was treated with TSA and Zeb before being treated with TRAIL (TZT) to compare the effect on MDA-MB-231 and MCF-7. The cell viability, cell cycle and migration assays were conducted on these cells, prior to reverse-transcription-polymerase chain reaction (RT-PCR) targeted on proliferating cell nuclear antigen (PCNA), cyclin-dependent kinase 2 (CDK2), matrix metalloproteinase 9 (MMP9). TZT induced a significant increase in G0/G1-arrested cell population and reduction in cell viability in 231-EGFP. These were verified by the suppression of PCNA and CDK2 mRNA expression. However, there was a negligible effect to reduce the cell migration of the invasive MDA-MB-231 and 231-EGFP cells in accordance with the lack of down-regulation of MMP9. In conclusion, this research shows that TSA and Zeb have sensitized breast cancer towards TRAIL treatment in 231-EGFP cells, validating the potentiality of E-cadherin as a biomarker of TRAIL treatment efficacy in the invasive breast cancer.

Molecular Docking Study of Astaxanthin Derived from Radio-Resistant Bacterium Deinococcus sp. Strain WMA-LM9 to Matrix Metalloproteinase-1, 3 (MMP-1, MMP-3)

Objective: To assess role of astaxanthin in downregulation of matrix metalloproteinases (MMP-1 and MMP-3) as potential mitigator for skin aging and antioxidant for different pathological diseasesStudy Design: Cross sectional.Place and Duration of Study: The study was carried out at Department of Biological Sciences of National University of Medical Sciences, Rawalpindi, Pakistan from June 2019 to March 2020.Materials and Methods: The docking studies of two different matrix metalloproteinases (MMP-1, MMP-3) with astaxanthin were carried out using Autodock/vina. The structural details were obtained from protein data bank and subjected to energy minimization using the UCSF Chimera 1.12.|Results: Out of the two selected targets, we found the highest binding energy (-11.9 kacl/mol) was for stromelysin-1 and astaxanthin docked complex. Astaxanthin was found to bind within the reported, predominantly hydrophobic S1, active site of protein mostly by hydrophobic interactions. The catalytic zinc ion has also shown to establish an electrostatic interaction with histidine residues of the MMPs.Conclusion: The current results suggest that astaxanthin has potential inhibitory activity on MMP-1 and MMP-3 and can be used as treatment to sunburns and skin aging.

Inhibition of the Epigenetic Reader BRD4 Reduces SIRPα-Mediated Phagocytosis and Melanoma Invasion

BRD4 acts as an epigenetic reader to regulate gene transcription. It represents a valid therapeutic target in cancer, and several selective and potent small molecule inhibitors have been discovered. A study by Le etal. (2020) published in Journal of Investigative Dermatology (2020) demonstrates that BRD4 inhibition reduces the invasive behavior of melanoma cells associated with matrix metalloproteinase-2 downregulation and increases phagocytosis by myeloid cells through SIRPα downregulation.

GOLPH3 Promotes Vasculogenic Mimicry via the Epithelial Mesenchymal Transition in Glioblastoma Cells.

To evaluate the relationship between Golgi phosphoprotein 3 (GOLPH3) and vasculogenic mimicry (VM) in glioblastoma cells. Glioma tissues from 40 glioma patients with different pathological grades were collected. GOLPH3 and VM were evaluated by immunostaining in glioma tissues. Then, the correlation between GOLPH3 and VM were analyzed clinically. Additionally, a GOLPH3-downregulation lentivirus was constructed and transfected into the human primary glioblastoma cell line, U-87 MG. Afterwards, apoptosis, migration and invasion were assessed to determine the effects of downregulation GOLPH3. Then, E-cadherin and matrix metalloproteinase 2 (MMP2) were detected for assessment of the epithelial mesenchymal transition (EMT). GOLPH3 and VM were found to be positively correlated following clinical analysis (p < 0.01, r=0.788). Furthermore, GOLPH3 downregulation suppressed the migration and invasion of U87 MG cells (p < 0.05), followed by upregulation of E-cadherin and downregulation of MMP2. Collectively, our results demonstrate that GOLPH3 promoted VM in glioblastoma cells and that the corresponding mechanism was associated with the EMT. Our finding suggests that GOLPH3 may represent a promising therapeutic target for mitigating the recurrence and invasion of gliomas.

The Anti-photoaging Effects of Pre- and Post-treatment of Platelet-rich Plasma on UVB-damaged HaCaT Keratinocytes.

To investigate the anti-photoaging effects of pre- and post-treatment of PRP on UVB-damaged HaCaT cells. HaCaT cells were irradiated with 80mJ/cm2 UVB, before or after PRP treatment (1000×107 /L), and following measurements were taken: survival rate of UVB-irradiated HaCaT cells, malondialdehyde (MDA) content and activities of glutathione peroxidase (GSH-Px), superoxide dismutase (SOD) and catalase (CAT). Western blot was used to determine the effect of different PRP intervention on the expression of PI3K, AKT, ERK, MMP-1, MMP-9, TIMP-1 and γ-H2AX in the UVB-irradiated HaCaT cells. pre- and post-PRP treatment reduced MDA content and increased the activities of GSH-Px, SOD and CAT in photoaged HaCaT cells. These changes resulted in reduced cytotoxic effects. Besides, different PRP intervention promoted cell proliferation via PI3K/AKT pathway. Furthermore, PRP application suppressed the expression of γ-H2AX. Also, PRP intervention alleviated photoaging effects by upregulating the expression level of tissue inhibitor of metalloproteinases-1 (TIMP-1) while downregulating matrix metalloproteinase (MMP) expression level in photoaged HaCaT cells. pre- and post-PRP treatment play anti-photoaging role through strengthening cellular oxidative defense capacity, mitigating MMP expression, alleviating DNA damages and promoting proliferation of UVB-irradiated HaCaT cells.

Alectinib and lorlatinib function by modulating EMT-related proteins and MMPs in NSCLC metastasis.

Most advanced non-small cell lung cancer (NSCLC) patients are accompanied by brain metastasis which is the major cause of increased mortality. The fusion rearrangement of anaplastic lymphoma kinase (ALK) gene is an important feature of brain metastasis in lung cancer. The novel ALK inhibitors alectinib and lorlatinib are shown to be effective against NSCLC brain metastasis, while their underlying mechanism of action is unclear. Epithelial–mesenchymal transition (EMT) proteins and matrix metalloproteinases (MMPs) play important roles in brain metastasis by regulating the blood-brain barrier (BBB). To reveal the molecular function of alectinib and lorlatinib, we explored their effects on the cellular levels of EMT markers: VIM and FN1 and the matrix metalloproteinases MMP-9 and MMP-7. The mRNA and protein levels of VIM, FN1, MMP-9, and MMP-7 were elevated in H3122 cells. However, upon alectinib and lorlatinib treatment, the levels were significantly reduced. Similar results were obtained when these experiments were performed either in a dose-dependent or time-dependent manner. Furthermore, alectinib and lorlatinib also inhibited the cell viability and migration of H3122 cells. Interestingly, in comparison to individual drugs, the combination of alectinib and lorlatinib was found to be substantially more effective. Overall, these results suggest that alectinib and lorlatinib possibly function through the downregulation of MMPs and EMT in NSCLC metastasis.

Apigenin as Tumor Suppressor in Cancers: Biotherapeutic Activity, Nanodelivery, and Mechanisms With Emphasis on Pancreatic Cancer.

Pancreatic cancer is the most lethal malignancy of the gastrointestinal tract. Due to its propensity for early local and distant spread, affected patients possess extremely poor prognosis. Currently applied treatments are not effective enough to eradicate all cancer cells, and minimize their migration. Besides, these treatments are associated with adverse effects on normal cells and organs. These therapies are not able to increase the overall survival rate of patients; hence, finding novel adjuvants or alternatives is so essential. Up to now, medicinal herbs were utilized for therapeutic goals. Herbal-based medicine, as traditional biotherapeutics, were employed for cancer treatment. Of them, apigenin, as a bioactive flavonoid that possesses numerous biological properties (e.g., anti-inflammatory and anti-oxidant effects), has shown substantial anticancer activity. It seems that apigenin is capable of suppressing the proliferation of cancer cells via the induction of cell cycle arrest and apoptosis. Besides, apigenin inhibits metastasis via down-regulation of matrix metalloproteinases and the Akt signaling pathway. In pancreatic cancer cells, apigenin sensitizes cells in chemotherapy, and affects molecular pathways such as the hypoxia inducible factor (HIF), vascular endothelial growth factor (VEGF), and glucose transporter-1 (GLUT-1). Herein, the biotherapeutic activity of apigenin and its mechanisms toward cancer cells are presented in the current review to shed some light on anti-tumor activity of apigenin in different cancers, with an emphasis on pancreatic cancer.

Metformin as a Potential Neoadjuvant Therapeutic Agent in Head and Neck Squamous Cell Carcinoma

Oral cancer is the sixth most common cancer worldwide with over 49 000 new cases expected in 2017 in the United States alone. Cancer of the oral cavity and oropharynx results in approximately 9700 deaths each year, with only a 50-60% 5-year survival rate. In addition to early diagnosis, research to develop new therapies targeted at tumor spread represent a potential strategy to lessen morbidity and mortality for those affected by oral squamous cell carcinoma (OSCC). Metformin is a widely used lipophilic biguanide that inhibits hepatic gluconeogenesis and improves peripheral utilization of glucose. Patients with type 2 diabetes treated with metformin have a lower risk of developing oral cancer when compared to those treated with drugs other than metformin. Metformin may also reduce locoregional recurrence (LRR) and improved overall survival and disease-free survival in patients with oral and hypopharyngeal cell carcinoma.1 Metformin has also been shown to reduce the size and number of carcinogen-induced oral tumoral lesions, preventing their spontaneous conversion to squamous cell carcinomas. Many anti-tumorigenic effects of metformin may be mediated through the activation of AMP-kinase (AMPK) and LKB1 and are summarized in Figure 1.3 Metformin activation of AMPK inhibits the mechanistic target of rapamycin (mTOR), a protein kinase that coordinates the eukaryotic cell growth and metabolism. Metformin controls the tumor microenvironment through the inhibition of matrix metalloproteinase production, decreasing tissue invasion. This study will include 3 oral tongue squamous cell carcinoma cell lines, 1 of them HPV16+. The cells were cultured and kept according to cell bank instructions. The cells were treated with a range of increasing doses of metformin (50μM-10mM) and left to incubate for 24 hours. The gel was washed and incubated in a cofactors buffer. The gels are then stained with Coomassie blue, de-stained, and imaged with AlphaView (ProteinSimple). For western blots, the same amount of total protein for each sample was run in a polyacrylamide gel, transferred into a nitrocellulose, and probed for signaling molecules and MMPs. Membranes were scanned with Image Studio Lite (LI-COR Biosciences). Additionally, to check mitochondrial activity, cells were incubated with metformin for 24 hours and then collected to be used for high-resolution spirometry. Respirometry data were analyzed using DatLab 7 (Oroboros O2k). Statistical analysis was performed in Prism 8 (GraphPad Software, San Diego, CA). Metformin decreased the expression of MMP-2 and MMP-9 in a dose dependent manner when compared to non-treated controls (n = 4, P < .001-0.0001, 2-way ANOVA, Dunnett’s multiple comparisons). Metformin also decreased the activity of MMP-2 and MMP-9 in a dose dependent manner when compared to non-treated controls (Figure 2). Respirometry data showed no significant different at therapeutically relevant doses of metformin (50-100μM) when compared to non-treated controls (n = 4, P > .5, 2-way ANOVA, Dunnett’s multiple comparisons). Effective chemoprevention is critical for improving outcomes of oral cancer. Metformin has been reported to exhibit chemopreventive properties, in vitro as well as in patients with oral cancer. In this study, the chemopreventive efficacy of Metformin is tested in the context of head and neck squamous cell carcinoma, effectively characterizing novel pathways of metastatic inhibition via downregulation of matrix metalloproteinases -2 and -9.

Gradient hydrogels for screening stiffness effects on patient-derived glioblastoma xenograft cellfates in 3D.

Brain cancer is a devastating disease given its extreme invasiveness and intricate location. Glioblastoma multiforme (GBM) is one of the most common forms of brain cancer, and cancer progression is often correlated with significantly altered tissue stiffness. To elucidate the effect of matrix stiffness on GBM cell fates, previous research is largely limited to 2D studies using immortalized cell lines, which has limited physiological relevance. The objective of the study is to develop gradient hydrogels with brain-mimicking stiffness range as a 3Din vitro GBM model for screening of the effects of matrix stiffness on GBM. To increase the physiological relevance, patient-derived tumor xenograft (PDTX) GBM cells were used. Our gradient platform allows formation of cell-containing hydrogels with stiffness ranging from 40 Pa to 1,300 Pa within a few minutes. By focusing on a brain-mimicking stiffness range, this gradient hydrogel platform is designed for investigating brain cancer. Increasing stiffness led to decreased GBM proliferation and less spreading, which is accompanied by downregulation of matrix-metalloproteinases (MMPs). Using temozolomide (TMZ) as a model drug, we demonstrate that increasing stiffness led to higher drug resistance by PDTX GBM cells in 3D, suggesting matrix stiffness can directly modulate how GBM cells respond to drug treatment. While the current study focuses on stiffness gradient, the setup may also be adapted for screening other cancer niche cues such as how biochemical ligand gradient modulates brain cancer progression and drug responses using reduced materials and time.

Glucagon-like peptide 1 treatment reverses vascular remodelling by downregulating matrix metalloproteinase 1 expression through inhibition of the ERK1/2/NF-κB signalling pathway

In addition to serving as an incretin-based treatment of type 2 diabetes mellitus (T2DM), glucagon-like peptide 1 (GLP-1) can also reverse cardiovascular diseases caused by vascular remodelling. However, a detailed mechanism underlying how GLP-1 reverses vascular remodelling remains unclear. Here, Spontaneous hypertension rats (SHR) were used as an in vivo model of vascular remodelling. Treatment with a GLP-1 receptor (GLP-1R) agonist Liraglutide or dipeptidyl peptidase 4 (DPP4) inhibitor Alogliptin decreased systolic blood pressure (SBP), diastolic blood pressure (DBP), thickness of vascular wall, and overall collagen levels in SHR. In vitro vascular remodelling can be induced by exposing rat aortic smooth muscle cells (RASMC) to angiotensin II (Ang II); GLP-1 treatment attenuated AngII induction of RASMC proliferation, migration, and excessive extracellular matrix (ECM) degradation. Downregulation of matrix metalloproteinase 1 (MMP1) enhanced the inhibitory effects of GLP-1, and extracellular regulated protein kinase 1/2 (ERK1/2) and nuclear factor kappa-B (NF-κB) signalling participated in these processes. These results provide a new mechanistic understanding of key therapeutic strategies for the treatment of vascular remodelling-related diseases.

PDLIM2 protects articular chondrocytes from lipopolysaccharide-induced apoptosis, degeneration and inflammatory injury through down-regulation of nuclear factor (NF)-κB signaling

Excessive inflammatory response-induced apoptosis and the degeneration of articular chondrocytes contribute to the development and progression of osteoarthritis. PDZ and LIM domain containing protein 2 (PDLIM2) has emerged as one of the pivotal regulators in orchestrating an inflammatory response through regulating the activity of transcription factor nuclear factor (NF)-κB. However, whether PDLIM2 participates in the articular chondrocyte-associated inflammatory response in osteoarthritis remains unknown. In the current study, we aimed to explore the biological function of PDLIM2 in lipopolysaccharide (LPS)-stimulated articular chondrocytes, an in vitro model of osteoarthritis. Herein, we found that PDLIM2 expression was significantly down-regulated in chondrocytes in response to LPS exposure. Functional experiments revealed that PDLIM2 overexpression increased the viability and decreased the apoptosis of chondrocytes following LPS treatment. Moreover, PDLIM2 overexpression attenuated LPS-induced degeneration of chondrocytes via the down-regulation of matrix metalloproteinase (MMP)-3 and MMP-13 and the up-regulation of COL2A1 and ACAN. In addition, the overexpression of PDLIM2 decreased LPS-induced production of interleukin (IL)-1β, IL-6 and TNF-α. In contrast, depletion of PDLIM2 exhibited the opposite effect. Mechanism research elucidated that PDLIM2 repressed the activation of NF-κB signaling associated with the down-regulation of NF-κB p65 protein expression. PDLIM2 depletion-exacerbated LPS-induced injury was significantly reversed by NF-κB inhibition. Taken together, these results demonstrate that PDLIM2 overexpression attenuates LPS-induced injury of articular chondrocytes through the inactivation of NF-κB signaling.

Antifibrotic effect of curcumin, N-acetyl cysteine and propolis extract against bisphenol A-induced hepatotoxicity in rats: Prophylaxis versus co-treatment

AimsBisphenol A (BPA) has been shown to induce liver fibrosis in rodents. Therefore, this study examined the protective effect of a triple combination of curcumin (Cur), N-acetyl cysteine (NAC) and propolis (Prp) extract against BPA-induced hepatic fibrosis. Methods100 Wistar male rats were equally assigned into 10 groups; one group was designated as control. 10 rats were gavaged with BPA (50 mg/kg/day) for 8 wk and left un-treated (BPA group). The remaining 80 rats were divided into 8 groups, distributed in 2 models. Protective model: rats were daily co-treated with BPA and Cur (100 mg/kg, p.o) or NAC (150 mg/kg, p.o) or Prp (200 mg/kg, p.o) or their combination for 8 wk. Preventive model: rats were daily treated with Cur or NAC or Prp or their combination for 4 wk before BPA administration and then in the same manner as protective model. Key findingsCurrent treatment interventions significantly alleviated BPA-induced hepatic damage and fibrosis. They also restored pro-oxidant/antioxidant balance, shifted cytokine balance towards the anti-inflammatory side, decreasing interleukin-1β/interleukin-10 ratio. Moreover, these compounds seem to exert anti-apoptotic effects by increasing the immunoexpression of B-cell lymphoma 2 in hepatocytes and decreasing hepatic caspase-3 content. Finally, they ameliorated extracellular matrix turn over through down-regulation of matrix metalloproteinase-9 and up-regulation of tissue inhibitor of matrix metalloproteinase-2 genetic expression. SignificanceCurrent treatments guarded against BPA-induced hepatic fibrosis due to their antioxidant, anti-inflammatory and anti-apoptotic properties, decreasing extracellular matrix turnover. Interestingly, the triple therapy provided hepatoprotection superior to monotherapy. Besides, prophylactic and concurrent treatments seem to be more effective than concurrent treatments.

High expression of FUSE binding protein 1 in breast cancer stimulates cell proliferation and diminishes drug sensitivity.

Breast cancer is the most common malignant tumor affecting women worldwide and is divided into the following subtypes: Luminal A, Luminal B, HER-2 overexpression and triple-negative breast cancer (TNBC). TNBC accounts for approximately 15-20% of all breast cancer cases. Due to the characteristics of low differentiation, the likelyhood of recurrence and metastasis, strong invasiveness and the lack of hormone receptors and human epidermal growth factor receptor 2 (HER2), patients with TNBC cannot benefit from endocrine therapy or other available targeted agents. Chemotherapy is one of the main treatments for patients with TNBC, and cisplatin is one of the most commonly used and effective drugs. The human far upstream element binding protein 1 (FBP1) is a potent pro-proliferative and anti-apoptotic oncoprotein, which is overexpressed in numerous tumor types. The present study demonstrated that FBP1 and its target, c-Myc, were more highly expressed in breast cancer tissues compared with para-carcinoma tissues, and the FBP1 and c-Myc levels are decreased by cisplatin treatment. The knockdown of FBP1 in TNBC cells decreased cell proliferation by arresting the cell cycle at the G2 phase. The knockdown of FBP1 decreased the expression of G2 phase-associateed protein cyclin A2, whereas it increased that of cyclin B1 and p-CDC2. Furthermore, the knockdown of FBP1 decreased cell migration and metastasis by downregulating matrix metalloproteinase 2 expression, and enhanced the sensitivity of TNBC cells to cisplatin by inducing apoptosis. These results thus suggest that FBP1 is a potential novel biological marker for the diagnosis and treatment of TNBC.

MiR-183-5p suppressed the invasion and migration of HTR-8/SVneo trophoblast cells partly via targeting MMP-9 in preeclampsia.

Preeclampsia (PE), a common obstetrical disorder, is characterized by impaired migration and invasion abilities of trophoblastic cells. MicroRNA-183-5p (miR-183) was reported to regulate cell migration and invasion in various types of human cancers; however, its role in the pathogenesis of PE remains elusive. Herein, we investigated the role of miR-183 in HTR-8/SVneo trophoblast cells invasion and migration and explored the underlying mechanism. Our results showed that miR-183 was significantly up-regulated in placental tissues from pregnant women compared with that in normal pregnant women. Overexpression of miR-183 inhibited proliferation, migration and invasion, as well as induced apoptosis in HTR-8/SVneo cells. Otherwise, down-regulation of miR-183 achieved the opposite effects. Bioinformatics prediction and luciferase reporter assay confirmed that matrix metalloproteinase-9 (MMP-9) is a target of miR-183. In addition, MMP-9 expression was significantly down-regulated, and inversely correlated with the miR-183 level in placental tissues from pregnant women with severe PE. Down-regulation of MMP-9 suppressed the trophoblast cell invasion and migration, whereas overexpression of MMP-9 promoted cell invasion and migration in HTR-8/SVneo cells. More importantly, up-regulation of MMP-9 reversed the inhibitory effects of miR-183 on cell invasion and migration in trophoblast cells. Collectively, our findings suggested that miR-183 may play critical roles in the pathogenesis of PE and serve as a potential biomarker for severe PE.

Inhibitory effects of brusatol delivered using glycosaminoglycan‑placental chondroitinsulfateA‑modified nanoparticles on the proliferation, migration and invasion of cancer cells.

Breakthroughs in cancer management result from the development of drugs that can be used for early diagnosis and effective treatment. Surgery, chemotherapy, radiotherapy and hormone therapy are the main anticancer therapies. However, traditional cancer chemotherapy is associated with serious systemic side effects. Nanoparticles (NPs) provide an effective solution for cancer treatment via the targeted delivery of drugs to cancer cells, while minimizing injury to normal cells. Glycosaminoglycan-placental chondroitin sulfate A (plCSA) is expressed in a number of tumor cells and trophoblasts. A plCSA-binding peptide (plCSA-BP) was isolated from malaria protein VAR2CSA, which can effectively promote the binding of lipid polymer NPs to tumor cells, thereby significantly enhancing the anticancer effect of encapsulated drugs. Brusatol is an important compound derived from Brucea javanica that exerts a multitude of biological effects, including inhibiting tumor cell growth, reducing the reproduction of malaria parasites, reducing inflammation and resisting virus invasion. In the present study, brusatol-loaded NPs (BNPs) or coumarin 6 NPs (CNPs), plCSA-BP and scrambled control peptide-bound BNPs or CNPs were prepared. Ovarian cancer cells (SKOV3), endometrial cancer cells (HEC-1-A) and lung cancer cells (A549) were treated with the NPs. The uptake of plCSA-CNPs by tumor cells was found to be markedly higher compared with that of other types of NPs. Further studies demonstrated that the plCSA-BNPs promoted the apoptosis of cancer cells more effectively and inhibited their proliferation, invasion and migration, accompanied by downregulation of matrix metalloproteinase (MMP)-2, MMP-9 and B-cell CLL/lymphoma 2 (BCL2) levels, but upregulation of BCL2-associated X protein BAX and cleaved caspase-3 levels. The results demonstrated the potential of brusatol delivered by plCSA-modified NPs as a chemotherapeutic agent for the targeted therapy of tumors by regulating the BCL2, BAX, cleaved caspase-3, MMP-2 and MMP-9 pathways, and indicated that it may be an effective and safe strategy for the treatment of various tumors.

Quercetin Improves Inflammation, Oxidative Stress, and Impaired Wound Healing in Atopic Dermatitis Model of Human Keratinocytes.

Background: Atopic dermatitis (AD) is a common inflammatory skin disease with complex pathogenesis. Natural flavonoids exhibit strong anti-inflammatory and antioxidant properties in many human diseases. In this study, the potential bioactive effect of quercetin, a polyphenolic plant-derived flavonoid, on the AD model of human keratinocytes was evaluated. Methods: Immortalized human HaCaT keratinocytes were treated with interleukin (IL) -4, -13, and tumor necrosis factor-α to mimic AD features in vitro. Then effects of quercetin on inflammation, oxidative stress, and wound healing were assessed. Results: Pretreatment of the cells with 1.5 μM of quercetin significantly reduced the expression of AD-induced IL-1β, IL-6, IL-8, and thymic stromal lymphopoietin, while it strongly enhanced the expression of superoxide dismutase-1 (SOD1), SOD2, catalase, glutathione peroxidase, and IL-10. Quercetin promoted wound healing by inducing epithelial-mesenchymal transition, which was supported by the upregulation of Twist and Snail mRNA expression. Unexpectedly, quercetin pretreatment of AD-induced cells upregulated the mRNA expression of occludin and E-cadherin, while downregulating matrix metalloproteinase 1 (MMP1), MMP2, and MMP9 expression. The pretreatment inhibited AD-induced phosphorylation of extracellular signal-regulated kinase 1/2/mitogen-activated protein kinase (ERK1/2 MAPK) and the expression of nuclear factor-kappa B (NF-κB), but it did not alter signal transducer and activator of transcription 6 (STAT6) phosphorylation. Conclusion: Quercetin may serve as a potential bioactive substance for atopic dermatitis-related symptoms through anti-inflammatory and antioxidant activities along with its acceleration of wound healing via ERK1/2 MAPK and NF-κB pathways.

Features of Stable Plaque Phenotype Are Increased by Macrophage Specific Insulin‐Like Growth Factor 1

Insulin‐like growth factor I (IGF‐1) reduces atherosclerosis in Apoe‐null mice and promotes stable plaque phenotype. We reported that deficiency for macrophage (MF)‐specific IGF‐1 receptor upregulated matrix metalloproteinases (MMPs) and promoted atherogenesis. To test whether MF‐specific IGF‐1 will suppress MMPs, decrease atherosclerosis, and change plaque stability, we generated Apoe‐null mice with IGF‐1 overexpression driven by MF‐specific promoter (scavenger receptor A) (SRA12 and SRA17 mice). IGF‐1 level was increased in peritoneal MF isolated from SRA12 (21±4% increase) and from SRA17 mice (52±9% increase) compared to control Apoe‐null mice (both P&lt;0.05). SRA12 and SRA17 mice were fed with a high fat diet for 12 weeks. MF‐specific IGF‐1 overexpression significantly decreased atherosclerotic burden (SRA12, 23±3% decrease, n=20, and SRA17, 28±6% decrease, n=16, vs. control, n=29, en face analysis) and increased plaque collagen levels (11.1±6%, SRA12 and 66.5±11%, n=11, SRA17, Trichrome). Atherosclerotic plaques in both SRA12 and SRA17 mice had reduced necrotic cores suggesting elevated plaque stability. Increased levels of plaque collagen in SRA17 mice correlated with a significant decrease of MMP1, MMP2, MMP8, MMP12, MMP13, and MMP14 protein levels in peritoneal MF (immunoblotting, 30–72% decrease vs. control). To assess monocyte recruitment into the plaque, macrophage‐cleavable fluorescent red beads were injected (IV). Plaques in both SRA12 and SRA17 mice have reduced number of red beads compared to Apoe‐null mice (77±2% decrease, p=0.027, 74±3% decrease, p=0.0592, respectively) suggesting decreased monocyte recruitment. Cutlured human THP‐1 MFs were incubated with IGF‐1 (0, 20, 100ng/mL) for 48 hrs. IGF‐1 (20ng/ml) decreased expression of MMP8 (34±13% decrease, P&lt;0.05) and MMP14 protein (35±17% decrease, P&lt;0.05) (ELISA). To demonstrate that downregulation of MMPs was mediated via IGF1R‐dependent mechanism, we used Picropodophyllin (PPP), a specific IGF1R inhibitor. MF pre‐treatment with PPP (1ug/ml, 2hrs) abolished IGF‐1‐induced MMP8 downregulation in MF. In summary, we show that IGF‐1 reduced MMPs in vitro and MF IGF‐1 decreased MMPs, upregulated collagen and suppressed atherosclerotic burden in vivo. We identified the novel MMP‐dependent mechanism potentially mediating MF IGF‐1 atheroprotective effect by increasing features of stable plaque phenotype.Support or Funding InformationNIH/NHLBI 2R01HL070241‐13A1

Dysregulated Kisspeptin/Receptor Expression at the Maternal‐Fetal Interface in the Preeclamptic‐like BPH/5 Mouse

Preeclampsia (PE) is a hypertensive pregnancy syndrome estimated to cause the death of approximately 63,000 women worldwide annually. Insufficient invasion of extravillous trophoblast cells (EVT) into the maternal decidua and inadequate placentation are key adverse events in the pathogenesis of PE. Kisspeptins (KP), a family of small peptides derived from the kiss1 gene product, have been implicated as physiologic inhibitors of EVT invasion, a mechanism associated with upregulation of Tissue Inhibitor of Metalloproteinases (TIMP) and downregulation of Matrix Metalloproteinases (MMP). In PE, placental dysregulation of kiss1 and the KP receptor (kiss1r) have been proposed. Based on in vitro studies and evaluation of term PE placentas, it has been suggested that kiss1 and kiss1r mRNA are overexpressed at the maternal‐fetal interface during early gestation and excessively impair EVT invasion. However, the innate challenges involving human sample collection and early diagnosis of PE prevent a holistic understanding of KP role. Therefore, the objective of this study was to utilize the preeclamptic‐like BPH/5 mouse to investigate uteroplacental KP dysregulation throughout gestation. We have previously shown that pregnant BPH/5 females spontaneously develop poor placentation with decreased trophoblast invasion and inadequately remodeled decidual vessels. Hence, we hypothesized that uteroplacental expression of kiss1/kiss1r would be temporally dysregulated in the BPH/5 compared to the control C57 mice. Furthermore, we hypothesized that higher kiss1/kiss1r levels would be associated with upregulation of TIMP2, an important endogenous inhibitor of MMPs and cellular migration. To test our hypothesis, implantation sites and placentas of BPH/5 and C57 mice were collected at the embryonic days (e) 4.5, 7.5, 12.5, 14.5 and 18.5. The kiss1, kiss1r and TIMP2 mRNA levels were measured using RT‐PCR and comparisons were performed using unpaired t‐test. A similar expression pattern of kiss1 and kiss1r was seen throughout gestation in BPH/5 and control groups, with levels increasing from early (e4.5) to mid‐gestation (e14.5) and decreasing thereafter (e14.5 and e18.5). During the peak of decidualization (e7.5), BPH/5 mice presented 2‐fold higher kiss1 and kiss1r compared to C57 mice (n = 6; p ≤ 0. 05). In accordance with the proposed KP role, a 10‐fold increase in TIMP2 was also seen in BPH/5 mice at e7.5 (n = 6, p ≤ 0.05). In mid‐gestation (e14.5) kiss1r levels were 2‐fold lower in BPH/5 (n = 5) than in C57 (n = 6; p &lt; 0.05), whereas kiss1 levels were not different among groups (n = 6; p &gt; 0.05). In late gestation (e18.5), kiss1 and kiss1r were 3‐ and 2.5‐fold lower respectively in placentas of BPH/5 mice (n = 6; p ≤ 0.05). To the author’s knowledge, this is the first report of translational studies using an animal model of PE to investigate the role of KP in PE‐associated poor placental development. A kiss1/kiss1r dysregulation is reported, with higher expression levels in the preeclamptic‐like mouse in the peak of decidualization, and concurrent upregulation of TIMP2. Further studies are needed to investigate KP‐related upstream and downstream signaling pathways in the context of PE.