WISH Cells Research Articles

Role of the GalNAc-galectin pathway in the healing of premature rupture of membranes

BackgroundPremature rupture of the membranes (PROM) is a key cause of preterm birth and represents a major cause of neonatal mortality and morbidity. Natural products N-acetyl-d-galactosamine (GalNAc), which are basic building blocks of important polysaccharides in biological cells or tissues, such as chitin, glycoproteins, and glycolipids, may improve possible effects of wound healing.MethodsAn in vitro inflammation and oxidative stress model was constructed using tumor necrosis-α (TNF-α) and lipopolysaccharide (LPS) action on WISH cells. Human amniotic epithelial cells (hAECs) were primarily cultured by digestion to construct a wound model. The effects of GalNAc on anti-inflammatory and anti-oxidative stress, migration and proliferation, epithelial-mesenchymal transition (EMT), glycosaminoglycan (GAG)/hyaluronic acid (HA) production, and protein kinase B (Akt) pathway in hAECs and WISH cells were analyzed using the DCFH-DA fluorescent probe, ELISA, CCK-8, scratch, transwell migration, and western blot to determine the mechanism by which GalNAc promotes amniotic wound healing.ResultsGalNAc decreased IL-6 expression in TNF-α-stimulated WISH cells and ROS expression in LPS-stimulated WISH cells (P < 0.05). GalNAc promoted the expression of Gal-1 and Gal-3 with anti-inflammatory and anti-oxidative stress effects. GalNAc promoted the migration of hAECs (50% vs. 80%) and WISH cells through the Akt signaling pathway, EMT reached the point of promoting fetal membrane healing, and GalNAc did not affect the activity of hAECs and WISH cells (P > 0.05). GalNAc upregulated the expression of sGAG in WISH cells (P < 0.05) but did not affect HA levels (P > 0.05).ConclusionsGalNAc might be a potential target for the prevention and treatment of PROM through the galectin pathway, including (i) inflammation; (ii) epithelial-mesenchymal transition; (iii) proliferation and migration; and (iv) regression, remodeling, and healing.

N-allyl quinoxaline derivative exhibited potent and selective cytotoxicity through EGFR/VEGFR-mediated apoptosis: In vitro and in vivo studies.

The cytotoxic activity, EGFR/VEGFR2target inhibition, apoptotic activity, RT-PCR gene expression, in vivo employing a solid-Ehrlich carcinoma model, and in silico investigations for highlighting the binding affinity of eight quinoxaline derivatives were tested for anticancer activities. The results showed that compound 8 (N-allyl quinoxaline) had potent cytotoxicity against A594 and MCF-7 cancer cells with IC50 values of 0.86and 1.06 µM, respectively, with noncytotoxic activity against WISH and MCF-10A cells having IC50 values more than 100 µM. Furthermore, it strongly induced apoptotic cell death in A549 and MCF-7 cells by 43.13% and 34.07%, respectively, stopping the cell cycle at S and G1-phases. For the molecular target, the results showed that compound 8 had a promising EGFR inhibition activity with an IC50 value of 0.088 µM compared to Sorafenib (IC50 = 0.056 µM), and it had a promising VEGFR2 inhibition activity with an IC50 value of 0.108 µM compared to Sorafenib (IC50 = 0.049 µM). Treatment with compound 8 ameliorated biochemical and histochemical parameters near normal in the in vivo investigation, with a tumor inhibition ratio of 68.19% compared to 64.8% for 5-FU treatment. Finally, the molecular docking study demonstrated the binding affinity through binding energy and interactive binding mode inside the EGFR/VEGFR2 proteins. Potent EGFR and VEGFR2 inhibition of compound 8 suggests its potential for development as a selective anticancer drug.

Exploring pyrrolidinyl-spirooxindole natural products as promising platforms for the synthesis of novel spirooxindoles as EGFR/CDK2 inhibitors for halting breast cancer cells.

Cancer represents a global challenge, and the pursuit of developing new cancer treatments that are potent, safe, less prone to drug resistance, and associated with fewer side effects poses a significant challenge in cancer research and drug discovery. Drawing inspiration from pyrrolidinyl-spirooxindole natural products, a novel series of spirooxindoles has been synthesized through a one-pot three-component reaction, involving a [3 + 2] cycloaddition reaction. The cytotoxicity against breast cancer cells (MCF-7 and MDA-MB-231) and safety profile against WISH cells of the newly developed library were assessed using the MTT assay. Compounds 5l and 5o exhibited notable cytotoxicity against MCF-7 cells (IC50 = 3.4 and 4.12μM, respectively) and MDA-MB-231 cells (IC50 = 8.45 and 4.32μM, respectively) compared to Erlotinib. Conversely, compounds 5a-f displayed promising cytotoxicity against MCF-7 cells with IC50 values range (IC50 = 5.87-18.5μM) with selective activity against MDA-MB-231 cancer cells. Compound 5g demonstrated the highest cytotoxicity (IC50 = 2.8μM) among the tested compounds. Additionally, compounds 5g, 5l, and 5n were found to be safe (non-cytotoxic) against WISH cells with higher IC50 values ranging from 39.33 to 47.2μM. Compounds 5g, 5l, and 5n underwent testing for their inhibitory effects against EGFR and CDK-2. Remarkably, they demonstrated potent EGFR inhibition, with IC50 values of 0.026, 0.067, and 0.04μM and inhibition percentages of 92.6%, 89.8%, and 91.2%, respectively, when compared to Erlotinib (IC50 = 0.03 μM, 95.4%). Furthermore, these compounds exhibited potent CDK-2 inhibition, with IC50 values of 0.301, 0.345, and 0.557μM and inhibition percentages of 91.9%, 89.4%, and 88.7%, respectively, in contrast to Roscovitine (IC50 = 0.556 μM, 92.1%). RT-PCR analysis was performed on both untreated and 5g-treated MCF-7 cells to confirm apoptotic cell death. Treatment with 5g increased the gene expression of pro-apoptotic genes P53, Bax, caspases 3, 8, and 9 with notable fold changes while decreasing the expression of the anti-apoptotic gene Bcl-2. Molecular docking and dynamic simulations (100 ns simulation using AMBER22) were conducted to investigate the binding mode of the most potent candidates, namely, 5g, 5l, and 5n, within the active sites of EGFR and CDK-2.

Biobran-loaded core/shell nanofibrous scaffold: a promising wound dressing candidate.

This research examined the effectiveness of Biobran as a bioactive substance that could potentially improve wound healing. It also looked at how Biobran affects the properties of a nanofibrous scaffold made through coaxial electrospinning. This is the first study exploring the use of Biobran in this context and its interaction with nanofibrous scaffolds. The scaffolds were composed of poly(ε-caprolactone) (PCL) in the shell and various concentrations of Biobran blended with polyvinyl alcohol (PVA) in the core. The properties of the scaffolds were characterized by SEM, TEM, FTIR, XRD, TGA, DSC, stress-strain test, WCA, release test, MTT cytotoxicity assay, wound scratching assay, and the dye exclusion method using trypan blue. The scaffolds loaded with Biobran exhibited a more compact and smooth morphology compared with the scaffold without Biobran. The physical interaction and crystallinity of the polymers in the scaffolds were also affected by Biobran in a concentration-dependent manner. This positively influenced their tensile strength, elongation at break, thermal stability, and hydrophilicity. The porosity, water uptake capacity, and WVTR of the nanofibrous scaffolds are within the optimal ranges for wound healing. The release rate of Biobran, which revealed a biphasic release pattern, decreased with increasing Biobran concentration, resulting in controlled and sustained delivery of Biobran from the nanofiber scaffolds. The cell viability assays showed a dose-dependent effect of Biobran on WISH cells, which might be attributed to the positive effect of Biobran on the physicochemical properties of the nanofibrous scaffolds. These findings suggest that Biobran-loaded core/shell nanofiber scaffolds have a potential application in wound healing as an ideal multifunctional wound dressing.

Comparison of crude nano-hydroxyapatite extracted from calcified seaweed in terms of composition, antibacterial activity, and cytotoxicity

The objectives of this study were to synthesize nano-hydroxyapatite (nHAp) from red seaweed by an optimized and environmentally friendly thermal and chemical treatment method using minimal chemicals. Calcified tissues from seaweed were thermally treated to obtain calcium carbonate, which reacted with potassium dihydrogen phosphate to synthesize nHAp. Nano-hydroxyapatite materials, nHAp-Jr and nHAP-Co were produced from the seaweeds Jania rubens and Corallina officinalis, respectively. Results showed that the developed method successfully produced high yields of nHAp-Jr and nHAp-Co powders (>78 %). The crude composition of the synthesized nHAp-Jr and nHAp-Co materials was characterized using X-ray diffraction (XRD), Fourier transform infrared (FTIR), scan electron microscopic (SEM), Energy dispersive X-ray spectroscopy (EDAX), Brunauer-Emmett-Teller (BET), Barrett-Joyner-Halenda (BJH), and thermal gravimetric analysis (TGA). XRD patterns confirmed the formation of nHAp and calcium phosphate phases. SEM images showed the nanoparticle size of nHAp-Jr and nHAp-Co with an average distribution of 16.70 ± 0.71 nm. EDAX spectra identified the relative similar elemental compositions of nHAp-Jr and nHAp-Co. BET and BJH patterns revealed relatively similar specific surface area, total pore volume and average pore diameter for mesoporous size nHAp powders (2–50 nm). Antibacterial activity of nHAp-Jr and nHAp-Co showed good efficacy against the caries-causing agent, Streptococcus mutans compared to ciprofloxacin using the modified XTT assay. Cytotoxicity testing of nHAp-Jr and nHAp-Co against WISH cell line using modified MTT assay confirmed the relevant antibacterial effects without any side effect on hepatocytes. This sustainable method produced nHAp compounds from seaweed that show promising antibacterial properties and biocompatibility, highlighting the potential for biomedical applications.

Cytotoxicity and Apoptosis-Induction in MCF-7 Cells for New Pd(II) Complex Based on s-Triazine Ligand: Synthesis, Single Crystal X-ray Diffraction Analysis and Structural Investigations

The synthesis and X-ray structure analysis of the new [PdLCl2]*0.5 CH2Cl2 complex where L is hydrazono-s-triazine di-morpholine derivative, were presented. In the neutral inner sphere of this complex, the organic ligand L is acting as a NN-bidentate chelate via the pyridine and hydrazone N-atoms. The coordination configuration of the Pd(II) is completed by two chloride ions at cis-positions. The tetra-coordinated Pd(II) showed a distorted square planar geometry. The outer sphere comprised half methylene chloride molecule per [PdLCl2] as crystal solvent. The crystal stability is dominated by a number of weak C-H…N, C-H…Cl, and C-H…O non-covalent interactions. Based on Hirshfeld analysis, the H…H, N…H, H…Cl, O…H, Pd…C, and Cl…C intermolecular interactions contributed by 45.2, 9.3, 21.5, 5.8, 2.3, and 3.4%, respectively. DFT studies revealed closed shell characters for the Pd-N and Pd-Cl coordinate bonds. The net charge of Pd is also predicted to be 0.311 e and the amount of electron density transferred from the ligand groups is 1.689 e. The Pd(II) complex exhibited potent cytotoxic activity against MCF-7, HepG2, and A549 cells with IC50 values of 1.18, 4.74, and 5.22 μg/mL, compared to cisplatin with IC50 values of 4.1, 9.7, and 12.3 μg/mL, respectively. Additionally, it exhibited poor cytotoxicity against WISH cells with much higher IC50 values (IC50 = 37.2 μg/mL). Investigating apoptosis-induction, the Pd(II) complex induced apoptotic cell death by an 11-fold change in MCF-7 cells arresting the cell phase at the G0–G1 phase. Accordingly, Pd(II) complex can be developed as a promising anti-breast cancer agent.

A New Saponin (Zygo-albuside D) from Zygophyllum album Roots Triggers Apoptosis in Non-Small Cell Lung Carcinoma (A549 Cells) through CDK-2 Inhibition.

Phytochemical study of the ethyl acetate root extract of Zygophyllum album has resulted in the isolation of a new saponin, Zygo-albuside D (1), along with two known compounds; (3-O-[β-D-quinovopyranosyl]-quinovic acid) (2), which is first reported in the root, and catechin (3), first reported in the genus. Their chemical structures were established by NMR and high-resolution mass spectrometry (HRMS). The new saponin (1) exhibited promising cytotoxicity with IC50 values of 3.5 and 5.52 μM on A549 and PC-3 cancer cell lines, respectively, compared to doxorubicin with IC50 values of 9.44 and 11.39 μM on A549 and PC-3 cancer cell lines, respectively. While it had an IC50 value of 46.8 μM against WISH cells. Investigating apoptosis-induction, compound 1 induced total apoptotic cell death in A549 lung cancer cells by 32-fold; 21.53% compared to 0.67% in the untreated control cells. Finally, it upregulated the pro-apoptotic genes and downregulated the antiapoptotic gene using gene expression levels. Compound 1 exhibited remarkable CDK-2 target inhibition by 96.2% with an IC50 value of 117.6 nM compared to Roscovitine. The molecular docking study further confirmed the binding affinity of compound 1 as CDK2 and Bcl2 inhibitors that led to apoptosis induction in A549 cancer cells. Hence, this study highlights the importance of compound 1 in the design of a new anticancer agent with specific mechanisms.

Enhanced Expression of p53 and Suppression of PI3K/Akt/mTOR by Three Red Sea Algal Extracts: Insights on Their Composition by LC-MS-Based Metabolic Profiling and Molecular Networking.

Brown algae comprise up to 2000 species with wide dissemination in temperate zones. A comprehensive untargeted metabolic profiling guided by molecular networking of three uninvestigated Red-Sea-derived brown algae, namely Sirophysalis trinodis, Polycladia myrica, and Turbinaria triquetra, led to the identification of over 115 metabolites categorized as glycerolipids, fatty acids, sterol lipids, sphingolipids, and phospholipids. The three algae exhibited low-to-moderate antioxidant capacity using DPPH and ABTS assays. Preliminary in vitro antiproliferative studies showed that the algal extracts displayed high cytotoxic activity against a panel of cancer cell lines. The most potent activity was recorded against MCF-7 with IC50 values of 51.37 ± 1.19, 63.44 ± 1.13, and 59.70 ± 1.22 µg/mL for S. trinodis, P. myrica, and T. triquetra, respectively. The cytotoxicity of the algae was selective to MCF-7 without showing notable effects on the proliferation of normal human WISH cells. Morphological studies revealed that the algae caused cell shrinkage, increased cellular debris, triggered detachment, cell rounding, and cytoplasmic condensation in MCF-7 cancer cells. Mechanistic investigations using flow cytometry, qPCR, and Western blot showed that the algae induced apoptosis, initiated cell cycle arrest in the sub-G0/G1 phase, and inhibited the proliferation of cancer cells via increasing mRNA and protein expression of p53, while reducing the expression of PI3K, Akt, and mTOR.

Crude extract of J1 fermentation promotes apoptosis of cervical cancer cells

Abstract Cervical cancer is a typical cancer characterized by abnormal cell growth in the cervical area. Ginkgo biloba L. is a deciduous tree of the genus Ginkgo, possessing anti-cancer effects. The aim of this study was to explore the effect of strain J1 from Ginkgo biloba L. on apoptosis of cervical cancer cells. Bacteriostatic activity test, MTT assay and Flow cytometry were used in this study. Crude extract of J1 fermentation reduced cell growth in cervical cancer. The crude extract of the fermentation broth of strain J1 had a good inhibitory effect on Staphylococcus aureus. The crude extract of the J1 fermentation had no toxic effect on normal WISH cells in the range of anti-cervical cancer concentration. Crude extract of J1 fermentation induced apoptosis and regulated cell cycle in cervical cancer. The active compounds were separated and identified by preparative chromatography, and more than ten compounds were obtained. Our study suggests that the crude extract of J1 fermentation from Endophytic fungi of Ginkgo biloba reduced cell growth, and promoted apoptosis of cervical cancer, and is a potential therapeutic strategy for the treatment of cervical carcinoma.

Synthesis of silver and copper nanoparticle using Spirulina platensis and evaluation of their anticancer activity

ABSTRACT The present research displays the green synthesis of stable silver nanoparticles (Ag-NPs) and copper oxide nanoparticles (CuO-NPs). The aqueous solution of Spirulina platensis (blue green algae) source was used as a reducing and capping agent and this study assessed the cytotoxicity of Ag- and CuO-NPs on three cancer cell cultures: A549 (lung cancer), HCT (human colon cancer), Hep2 (laryngeal carcinoma cancer) and normal cell (WISH). For NPs characterization, the UV/Vis spectroscopy was used where their formation and crystallinity were proven with max values for Ag- and CuO-NPs of 425 and 234 nm, respectively. According to X-ray diffraction and transmission electron microscopy (TEM), Ag-NPs were spherical in shape (size 2.23–14.68 nm) and CuO-NPs were small (size 3.75–12.4 nm). Zeta potential analysis showed the particles potential, which was recorded by −14.95 ± 4.31 mV for Ag-NPs and −21.63 ± 4.90 mV for CuO-NPs. After that, Ag- and CuO-NPs were assessed for anticancer properties against A549, HCT, Hep2 and WISH. IC50 of Ag-NPs recorded 15.67, 12.94, 3.8 and 10.44 µg/ml for WISH, A549, HCT and Hep2, respectively. IC50 for CuO-NPs was recorded as 32.64, 54.59, 3.98 and 20.56 µg/ml for WISH, A549, HCT and Hep2 cells, respectively. Safety limits for WISH and A549 were achieved 98.64% by 2.44 µg/ml and 83.43% by 4.88 µg/ml of Ag-NPs, and it was found to be 97.94% by 2.44 µg/ml against HCT, while that for Hep2 is 95.9% by 2.44 µg/ml. Concerning the anticancer effect of CuO-NPs, the safety limit was recorded as 88.70% by 2.44 and 98.48% by 4.88 µg/ml against WISH and A549, while HCT reached 89.92% by 2.44 µg/ml and Hep2 was 83.33% by 4.88 µg/ml. Green nanotechnology applications such as Ag-NPs and CuO-NPs have numerous benefits of ecofriendliness and compatibility for biomedical applications such as anticancer effects against cancer cells.

In Vitro Cytotoxic Activity of Total Flavonoid from Equisetum Arvense Extract.

Normally happening substances like flavonoids are regarded as active candidates for the treatment and prevention of cancer The purpose of this study was to see how Iraqi E. arvense total flavonoid affected cell lines biologically and human lung fibroblast normal cell line (WISH). Plant powder was extracted by reflex apparatus, then thin-layer chromatography (TLC) was used to determine total flavonoids. Cytotoxicity assay (MTT) was used to determine the cytotoxic activity of the prepared plant against human breast cancer (MCF-7), cells human cervix cancer (HELA), human colon cancer (Caco-2) and human lung fibroblast normal cell line (WISH). The flavonoids Rutin, Quercetin, Kaempferol, and luteolin were detected using the Thin Layer Chromatography (TLC) technique. In contrast to the negative control, the extract inhibited cell growth to a highest of 82.158% for MCF-7 and 61.360% for Caco-2 at the concentration (100 µg/ml), and (54.880%) for Hela cell line at the concentration (100 µg/ml). In addition, the concentration (6.25 µg/ml) of total flavonoid extract produced a decrease in the growth of the normal WISH cell line to reach (1.094%). Equisetum arvense contain high amounts of flavonoids, the qualification of some flavonoids compounds was detected using TLC. The total flavonoids showed significant cytotoxic activity against various types of cancer cell lines and normal cell line in vitro, the antitumor activity was highly efficient in a dose and cell type dependent manner.

Synthesis, and Cytotoxic Activity of Novel Pyrazoline-Thiazolidinone Derivatives with Molecular Docking Studies

1-(4-substitutesd)-2-(5-phenyl-3-(1H-pyrrol-2-yl)-4,5-dihydropyrazol-1-yl)thiazol-5-yl)-2-aryldiazene 9a-f were prepared through reactions of 5-phenyl-3-(1H-pyrrol-2-yl)-4, 5-dihydropyrazole-1-carbothioamide 5 with hydrazonoyl halides. Furthermore, thioamide 5 was utilized as starting materials for thiazolidone synthesis 10 and arylidene thiazoles 15a–c. Also pyran derivatives 12a-b, 13a-b,14a-b and thioanilide derivaives 16 were obtained from the reaction of thiazilidone 10 with arylidenemalononitrile and phenyl isothiocyanat, respectively. When possible, the structures of experimentally synthesized compounds were identified by elemental analysis, spectral analysis, and alternative synthesis routes. Some of the synthesized compounds as 9f, 10, 12b, and 16 were screened for their cytotoxicity against MCF-7 and HCT-116 cells using the MTT assay. They exhibited remarkable cytotoxic activities with promising IC50 values. Interestingly, compound 16 exhibited potent cytotoxicity against MCF-7 and HCT-116 with IC50 values of 5.05 and 3.08 μM, compared to doxorubicin with IC50 values of 7.27 and 8.92 μM, respectively show interesting biological activities as anticancer activities. Additionally, the tested compounds were nontoxic against WISH cells with higher IC50 values. Hence, these compounds may serve as potent and selective cytotoxic agents.

Potential Serum Biomarkers Associated with Premature Rupture of Fetal Membranes in the First Trimester.

Premature rupture of the fetal membranes (PROM) is a common and important obstetric complication with increased risk of adverse consequences for both mothers and fetuses. An accurate and timely method to predict the occurrence of PROM is needed for ensuring maternal and fetal safety. Untargeted metabolomics was applied to characterize metabolite profiles related to PROM in early pregnancy. 41 serum samples from pregnant women who developed PROM later in gestation and 106 from healthy pregnant women as a control group, were analyzed. Logistic regression analysis was adjusted to analyze a PROM prediction model in the first trimester. A WISH amniotic cell viability assay was applied to explore the underlying mechanisms involved in PROM, mediated by C8-dihydroceramide used to mimic a potential biomarker (Cer 40:0; O2). Compared with healthy controls, 13 serum metabolites were identified. The prediction model comprising four compounds (Cer 40:0; O2, sphingosine, isohexanal and PC O-38:4) had moderate accuracy to predict PROM events with the maximum area under the curve of a receiver operating characteristics curve of approximately 0.70. Of these four compounds, Cer 40:0; O2 with an 1.81-fold change between PROM and healthy control serum samples was defined as a potential biomarker and inhibited the viability of WISH cells. This study sheds light on predicting PROM in early pregnancy and on understanding the underlying mechanism of PROM. Trial Registration: This study protocol has been registered at www.ClinicalTrials.gov, CT03651934, on 29 August 2018 (prior to recruitment).

Synthesis of Novel Phthalazinedione-Based Derivatives with Promising Cytotoxic, Anti-bacterial, and Molecular Docking Studies as VEGFR2 Inhibitors.

The parent ester methyl-3-[2-(4-oxo-3-phenyl-3,4-dihydro-phthalazin-1-yloxy)-acetylamino] has 18 compounds. The starting material for alkanoates, their corresponding hydrazides, hydrazones, and dipeptides were produced by chemoselective O-alkylation of 2-phenyl-2,3-dihydrophthalazine-1,4-dione with ethyl chloroacetate(4-oxo-3-phenyl-3,4-dihydro-phthalazin-1-yloxy) acetic acid methyl ester. The starting ester was hydrazinolyzed, then azide coupled with amino acid ester hydrochloride to produce several parent esters, and then hydrazinolyzed to produce parent hydrazides. These hydrazides were used to make a series of dipeptides by reacting them with amino acid ester hydrochloride under azide coupling conditions, and they were also condensed with a number of aldehydes to make the hydrazones. These derivatives were subjected to cytotoxicity against HCT-116 and MDA-MB-231 cells and anti-bacterial and molecular docking studies. Results indicated that the tested compounds, especially 7c and 8b with the phenyl phthalazinone moieties, had promising cytotoxicity against the HCT-116 cells with IC50 values of 1.36 and 2.34 μM, respectively. Additionally, the promising compounds 7c and 8b exhibited poor cytotoxicity against WISH cells with much higher IC50 values, so they were safe against normal cells. Compound 8c exhibited potent anti-bacterial activity with inhibition zones of 12 and 11 mm against Staphylococcus aureus and Escherichia coli, respectively. The molecular docking results of compounds 7c and 8b revealed a good binding disposition and the ligand–receptor interactions like the co-crystallized ligand of the VEGFR2 protein, which may be the proposed mode of action. Finally, compounds 7c and 8b exhibited good ADME pharmacokinetics with good drug-likeness parameters. Hence, detailed studies for the mechanism of action of such compounds are highly recommended for the development of new potent anti-cancer and anti-bacterial agents.

Development of a potential anti-cancer pulmonary nanosystem consisted of chitosan-doped LeciPlex loaded with resveratrol using a machine learning method

A novel self-assembled lecithin-based cationic nanocarrier (LeciPlex) doped with chitosan was prepared to improve Resveratrol solubility and anticancer efficacy. Using on a machine learning method based on regression analysis, LP5 (composed of lecithin, Peceol® and chitosan) was selected as the optimized one. This formula was spherical-shaped with a particle size of 59 nm, an encapsulation efficiency of 89% and a mean release time of 2.75 h together with a good mucoadhesive potential. Cascade impactor studies revealed the possible deep lung deposition of LP5 after nebulization. The safety of Resveratrol suspension and LP5 were checked on WISH cell line. Compared with Doxorubicin, both Resveratrol suspension and LP5 showed potent anticancer effects against the A549 cell line with IC50 values of 2.312, 2.200 and 1.708 μg/ml, respectively. These results indicated that the selected formula (LP5) significantly enhanced the crude drug anticancer effect by 63.2 fold and showed a higher anticancer selectivity index (>5.51). Furthermore, after 24 h, A549 up-took the LP5 more than the drug suspension by 40,000 fold. In addition, LP5 markedly induced the cellular apoptosis in A549 treated cells with a percentage of 73.1 compared to untreated cells. In conclusion, the results confirmed the enhanced lung anticancer effect, lower toxicity, and higher selectivity of the selected Resveratrol-loaded LeciPlex compared to the drug suspension. This study presents a potential nano-vesicular system for nebulization to combat lung cancer.

MiR-132-3p Modulates DUSP9-Dependent p38/JNK Signaling Pathways to Enhance Inflammation in the Amnion Leading to Labor.

Labor is a process of inflammation and hormonal changes involving both fetal and maternal compartments. MicroRNA-132-3p (miR-132-3p) has been reported to be involved in the development of inflammation-related diseases. However, little is known about its potential role in labor onset. This study aimed to explore the mechanism of miR-132-3p in amnion for labor initiation. In the mouse amnion membranes, the expression of miR-132-3p was found to increase gradually during late gestation. In human amniotic epithelial cell line (WISH), upregulation of miR-132-3p was found to increase proinflammatory cytokines and cyclooxygenase 2 (COX2) as well as prostaglandin E2 (PGE2), which was suppressed by miR-132-3p inhibitor. Dual-specificity phosphatase 9 (DUSP9) was identified as a novel target gene of miR-132-3p, which could be negatively regulated by miR-132-3p. DUSP9 was present in the mouse amnion epithelial cells, with a decrease in its abundance at 18.5 days post coitum (dpc) relative to 15.5 dpc. Silencing DUSP9 was found to facilitate the expression of proinflammatory cytokines and COX2 as well as PGE2 secretion in WISH cells, which could be attenuated by p38 inhibitor SB203580 or JNK inhibitor SP600125. Additionally, intraperitoneal injection of pregnant mice with miR-132-3p agomir not only caused preterm birth, but also promoted the abundance of COX2 as well as phosphorylated JNK and p38 levels, and decreased DUSP9 level in mouse amnion membranes. Collectively, miR-132-3p might participate in inflammation and PGE2 release via targeting DUSP9-dependent p38 and JNK signaling pathways to cause preterm birth.

Dexmedetomidine and LPS co-treatment attenuates inflammatory response on WISH cells via inhibition of p38/NF-κB signaling pathway.

BackgroundInflammatory dental diseases that occur during pregnancy can cause preterm labor and/or intrauterine growth restriction. Therefore, proactive treatment of dental diseases is necessary during pregnancy. Dexmedetomidine (DEX) is a widely used sedative in the dental field, but research on the effect of DEX on pregnancy is currently insufficient. In this study, we investigated the effects of co-treatment with DEX and lipopolysaccharide (LPS) on inflammatory responses in human amnion-derived WISH cells.MethodsHuman amnion-derived WISH cells were treated with 0.001, 0.01, 0.1, and 1 µg/mL DEX with 1 µg/mL LPS for 24 h. Cytotoxicity of WISH cells was evaluated by 3-(4,5-dimethylthiazol)-2,5-diphenyltetrazolium bromide (MTT) assay. The protein expression of cyclooxygenase-2 (COX-2), prostaglandin E2 (PGE2), p38, and nuclear factor kappa B (NF-κB) was examined by western blot analysis. The mRNA expression of pro-inflammatory cytokines such as interleukin (IL)-1β and tumor necrosis factor (TNF)-α was analyzed by real-time quantitative polymerase chain reaction.ResultsCo-treatment with DEX and LPS showed no cytotoxicity in the WISH cells. The mRNA expression of IL-1β and TNF-α decreased after co-treatment with DEX and LPS. DEX and LPS co-treatment decreased the protein expression of COX-2, PGE2, phospho-p38, and phospho-NF-κB in WISH cells.ConclusionCo-treatment with DEX and LPS suppressed the expression of COX-2 and PGE2, as well as pro-inflammatory cytokines such as IL-1β and TNF-α in WISH cells. In addition, the anti-inflammatory effect of DEX and LPS co-treatment was mediated by the inhibition of p38/NF-κB activation.

Propofol protects against lipopolysaccharide-induced inflammatory response in human amnion-derived WISH cells

BackgroundNonobstetric surgery is sometimes required during pregnancy, and neck abscess or facial bone fracture surgery cannot be postponed in pregnant women. However, dental surgery can be stressful and can cause inflammation, and the inflammatory response is a well-known major cause of preterm labor. Propofol is an intravenous anesthetic commonly used for general anesthesia and sedation. Studies investigating the effect of propofol on human amnion are rare. The current study investigated the effects of propofol on lipopolysaccharide (LPS)-induced inflammatory responses in human amnion-derived WISH cells.MethodsWISH cells were exposed to LPS for 24 h and co-treated with various concentrations of propofol (0.01–1 µg/ml). Cell viability was measured using the MTT assay. Nitric oxide (NO) production was analyzed using a microassay based on the Griess reaction. The protein expression of cyclooxygenase-2 (COX-2), prostaglandin E2 (PGE 2), p38, and phospho-p38 was analyzed using western blotting.ResultsPropofol did not affect the viability and NO production of WISH cells. Co-treatment with LPS and propofol reduced COX-2 and PGE2 protein expression and inhibited p38 phosphorylation in WISH cells.ConclusionPropofol does not affect the viability of WISH cells and inhibits LPS-induced expression of inflammatory factors. The inhibitory effect of propofol on inflammatory factor expression is likely mediated by the inhibition of p38 activation.

Spectral, structural and cytotoxicity studies on the newly synthesized n'1,n'3-diisonicotinoylmalonohydrazide and some of its bivalent metal complexes

The reaction of diethyl malonate with isonicotinic hydrazide resulted in the formation of a new ligand, N'1,N'3-diisonicotinoylmalonohydrazide (H4INM), which was characterized by elemental and spectral studies. The existence of the free ligand in keto form was shown by its spectral data. The ligand reacted with Cu(II), Co(II), Ni(II) and Zn(II) acetates to produce bimetallic complexes. The ligand chelated to the metal ion in binegative tetradentate fashion via malonyl carbonyl oxygen and N2H atoms except in Zn(II) complex, where it acted as neutral tetradentate through the isonicotinyl carbonyl oxygen and N1H atoms. The various spectral measurements revealed that all complexes had an octahedral geometry except the Cu(II) complex which has square planar structure supported by the ESR spectral data. Moreover, the DFT quantum chemical calculations were performed and the HOMO-LUMO energy gap of the ligand and complexes were comparable to the optical band gap (Eg) estimated using Tauc's equation. Furthermore, isolated compounds’ in vitro cytotoxic activity against Hela (Epithelioid Carcinoma) and WISH (Human amnion) cell lines of was investigated. According to the results, the ligand exhibits moderate and weak effect against the Hela and WISH cell lines, respectively. While, the metal complexes exhibited more potent effect against WISH than against Hela, with IC50 ranging from 23.65 ± 1.7 to 46.28 ± 3.4 and 56.36 ± 3.5 to 83.88 ± 4.7 µM, respectively.

The Role of IL-27 in the Systemic Inflammatory Response That Accompanies Preterm Labour.

This study aimed to investigate whether interleukin-27 (IL-27) activates maternal peripheral blood mononuclear cells (PBMCs) and induces inflammatory responses in amniotic epithelial cells in preterm labour (PL). The expression of IL-27p28, EBI3 and IL-27Rα was compared in maternal PBMCs of the PL, term labour (TL) and term not in labour (TNL) groups. The relationship between IL-27 and molecules associated with PBMC activation was investigated using bioinformatic and quantitative reverse transcription polymerase chain reaction (qRT-PCR) analyses. We investigated the inflammatory effects of IL-27 in PBMCs and its underlying mechanisms in vitro. In addition, we treated amniotic epithelial cells (WISH cells) with a PBMC-conditioned medium to identify the inflammatory effects of IL-27-treated PBMCs in amniotic epithelial cells. The expression of IL-27p28 and IL-27Rα in PBMCs of the PL group was higher than that in the TL/TNL groups. Bioinformatic analysis revealed that IL-27 was positively correlated with IFNG, IL6, IL1β, CXCL10 and ICAM1 in the whole blood samples of pregnant women in the PL group, which was confirmed using qRT-PCR. Furthermore, rhIL-27 promoted the expression of Th1 cell-related molecules (T-bet, IFN-γ and ICAM-1) and proinflammatory cytokines (IL-6 and IL-1β) in PBMCs in vitro, which was partially mediated by the JAK2/STAT1 pathway. In addition, it enhanced the expression of IL-27p28, EBI3 and IL-27Rα in PBMCs. Moreover, the expression of IL-6, IL-1β and TNF-α in WISH cells was significantly increased by the conditional medium derived from IL-27-treated PBMCs. IL-27 upregulated the expression of Th1 cell-related molecules and proinflammatory cytokines in PBMCs partially mediated by the JAK2/STAT1 pathway. Inflammatory responses were induced in WISH cells by a conditional medium derived from IL-27-treated PBMCs. Therefore, IL-27 may contribute to PL by promoting inflammation in maternal PBMCs and amniotic epithelial cells.