WI-38 Cells Research Articles

Phytochemical Analysis and Multifaceted Biomedical Activities of Nitraria retusa Extract as Natural Product-Based Therapies

This study examined the phytochemical profile and biomedical activities of Nitraria retusa, a halophytic and drought-resistant shrub. HPLC analysis showed gallic acid (1905.1 μg/g), catechin (1984.1 μg/g), and ellagic acid (2671.1 μg/g) as the primary constituents, while FT-IR analysis revealed a complex organic profile with significant functional groups. The extract demonstrated strong antioxidant activity in DPPH assays, outperforming ascorbic acid (IC50 = 18.7 ± 1.0 μg/mL) with an IC50 of 16.4 ± 4.4 μg/mL. It demonstrated specific antiproliferative effects on cancer cell lines as it showed selective cytotoxicity against cancer cell lines; normal WI38 cells were largely unaffected, showing 50.0% viability at 125 μg/mL. The most sensitive cell line was Caco2, which showed 50.0% viability at 125 μg/mL. Anti-diabetic properties were exhibited by means of inhibition of α-amylase (IC50 = 68.2 ± 4.2 μg/mL) and α-glucosidase (IC50 = 22.8 ± 3.3 μg/mL). Additionally, antimicrobial activity was observed to be broad-spectrum, and it was most effective against E. coli (32.6 mm inhibition zone at 400 μg/mL) and Penicillium glabrum (35.3 mm at 400 μg/mL). These findings highlight the potential of N. retusa in developing plant-based therapeutic approaches.

Design of a Magnetic Nanoplatform Based on CD26 Targeting and HSP90 Inhibition for Apoptosis and Ferroptosis-Mediated Elimination of Senescent Cells.

The accumulation of senescent cells, a hallmark of aging and age-related diseases, is also considered as a side effect of anticancer therapies, promoting drug resistance and leading to treatment failure. The use of senolytics, selective inducers of cell death in senescent cells, is a promising pharmacological antiaging and anticancer approach. However, more studies are needed to overcome the limitations of first-generation senolytics by the design of targeted senolytics and nanosenolytics and the validation of their usefulness in biological systems. In the present study, we have designed a nanoplatform composed of iron oxide nanoparticles functionalized with an antibody against a cell surface marker of senescent cells (CD26), and loaded with the senolytic drug HSP90 inhibitor 17-DMAG (MNP@CD26@17D). We have documented its action against oxidative stress-induced senescent human fibroblasts, WI-38 and BJ cells, and anticancer drug-induced senescent cutaneous squamous cell carcinoma A431 cells, demonstrating for the first time that CD26 is a valid marker of senescence in cancer cells. A dual response to MNP@CD26@17D stimulation in senescent cells was revealed, namely, apoptosis-based early response (2 h treatment) and ferroptosis-based late response (24 h treatment). MNP@CD26@17D-mediated ferroptosis might be executed by ferritinophagy as judged by elevated levels of the ferritinophagy marker NCOA4 and a decreased pool of ferritin. As 24 h treatment with MNP@CD26@17D did not induce hemolysis in human erythrocytes in vitro, this newly designed nanoplatform could be considered as an optimal multifunctional tool to target and eliminate senescent cells of skin origin, overcoming their apoptosis resistance.

Oridonin alleviates inflammation and endoplasmic reticulum stress in pediatric pneumonia via regulating the SIRT1-mediated Wnt/β-catenin signaling pathway.

Pediatric pneumonia is a prevalent and significant health concern worldwide, with elevated morbidity and mortality rates among affected children. This study was designed to elucidate the therapeutic impact of Oridonin (Ori) on pediatric pneumonia and unravel the underlying mechanisms involved. A pediatric infantile pneumonia model was established in mice through intratracheal administration of LPS. Additionally, a cell damage model was created in WI-38 cells by administering LPS. Protein levels were assessed via western blotting, and cell viability was measured with CCK-8. Inflammatory cytokines were quantified through ELISA, and specific assays were employed to evaluate oxidative stress markers. Flow cytometry was utilized to assess cell apoptosis. Ori alleviated lung inflammation, oxidative stress, apoptosis, and endoplasmic reticulum stress (ERS) in LPS-induced pneumonia mice. In addition, Ori increased the viability of LPS-induced pneumonia cells but decreased cell apoptosis. Furthermore, Ori reduced oxidative stress, inflammation, and ERS in LPS-induced pneumonia cells by enhancing SIRT1 to activate the Wnt/β-catenin pathway. This study suggested that Ori inhibited pediatric pneumonia by dampening the inflammatory response, oxidative stress, cell apoptosis, and ERS via the SIRT1/Wnt/β-catenin pathway.

Baicalin alleviates LPS-induced cytotoxicity in acute lung injury through mediating METTL14/SOX6 axis.

Baicalin (C21H18O11) is a flavonoid component extracted from scutellaria baicalensis with biological activity in various types of diseases, including acute lung injury (ALI). The relevant mechanism behind baicalin in ALI needs further investigation. ALI model in vitro was established by lipopolysaccharide (LPS) in WI-38 cells (lung fibroblast). Cell growth was determined via MTT assay and EdU assay. Apoptosis was assessed using flow cytometry, caspase 3 assay and TUNEL assay. Oxidative indicators and inflammatory cytokines were detected by commercial kits. Interaction between methyltransferase-like 14 (METTL14) and SRY-Box Transcription Factor 6 (SOX6) was studied using Methylated RNA Immunoprecipitation (MeRIP) and dual-luciferase reporter assay. RT-qPCR and western blot were applied for examining gene levels. Baicalin enhanced cell growth and reduced apoptosis, oxidative stress, inflammation after ALI was induced by LPS. Downregulation of SOX6 weakened LPS-induced cytotoxicity in WI-38 cells. Baicalin prevented from LPS-induced lung cell injury via reducing SOX6 expression. SOX6 expression was stabilized by METTL14 through its methylation modification. METTL14/SOX6 axis was related to the regulation of baicalin in LPS-treated WI-38 cells. Therefore, baicalin played an important role to inhibit LPS-induced cytotoxicity in vitro via METTL14-mediated methylation of SOX6.

Bio-computational modeling, POM analysis and molecular dynamic simulation for novel synthetic quinolone and benzo[d][1,3]oxazine candidates as antimicrobial inhibitors

The current study offers a metal-free, direct, and successful synthesis technique for a new series of quinolinone and benzo[d][1,3]oxazine, along with an assessment of their biological activities. Heteroannulation of anthranilic acid with carbonyl-containing chemicals (aroyl pyruvate, ethyl acetoacetatete, maleic anhydride, and ethyl cyanoacetate) resulted in the desired quinolones and benzo[d][1,3]oxazines. This technique introduces a number of fundamental breakthroughs in organic synthesis, including metal-free catalysts, smart reaction conditions with column purification, and a wide functional scope. Furthermore, the structure of the newly synthesized chemical series was investigated and validated using spectroscopic techniques. The synthesized series were evaluated for antibacterial (against gram-positive and gram-negative bacterial strains) and antifungal activity. The quinolone and benzo[d][1,3]oxazine candidates had remarkable antibacterial action. Furthermore, molecular docking investigations corroborated the biological studies using the Molecular Operating Environment and Petro Osiris Molinspiration (POM) experiments, which confirmed the activity of compounds 8, 15, and 17. Our studies on the cytotoxic activity of various chemicals have demonstrated that these compounds exhibit minimal toxicity. Specifically, when comparing the cytotoxic effects on human lung fibroblast (WI38) cells to those of Doxorubicin, a well-known chemotherapy agent, compounds 8, 15, and 17 showed weak cytotoxic effects on the normal WI38 cells. This indicates that these compounds may possess some level of selectivity and reduced toxicity towards normal cells, suggesting potential for further exploration as antibacterial agents with a safer profile for normal cells.

Diagnostic Value of lncRNA FGD5-AS1 Sponge miR-223-3p in Infantile Pneumonia and Its Prognostic Effect on Rehabilitation

Abstract Objective This study aimed to uncover the value of long noncoding RNA FGD5-AS1 (lncRNA FGD5-AS1) in the diagnosis of infantile pneumonia and explore its pathological mechanism in lung fibroblasts. This research may provide a potential biomarker for diagnosing patients and predicting their rehabilitation outcomes. Methods This study included 92 children with infantile pneumonia as the research object, and an equal number of healthy children were introduced as controls. The FGD5-AS1 content was detected using real-time quantitative polymerase chain reaction (RT-qPCR) assay. The diagnostic value of FGD5-AS1 was evaluated by receiver operating characteristic (ROC) and logistic analysis. The molecular mechanism of FGD5-AS1 and miR-223-3p was studied by luciferase activity assays. The impact of abnormal FGD5-AS1 expression on the proliferation and apoptosis of lung fibroblasts was analyzed using the cell counting kit-8 (CCK-8) and flow cytometry. Results FGD5-AS1 expression was decreased in the serum of infantile pneumonia patients, which may be a diagnostic marker for children with pneumonia. Furthermore, FGD5-AS1 has the ability to predict patient outcomes. FGD5-AS1 levels in lung fibroblasts (WI-38) decreased when induced by lipopolysaccharide (LPS). This decline resulted in reduced cell proliferation ability, increased apoptosis rate, and elevated inflammatory factor content. However, upregulated FGD5-AS1 counteracted the effects of LPS on WI-38 cells activity and inflammatory factors. Conclusion FGD5-AS1 may act as a potential marker in infantile pneumonia, and regulate the biological activity and inflammation level of lung fibroblasts by targeting miR-223-3p.

Involvement of cellular senescence in the effect of X-irradiated human lung fibroblast WI-38 cells on human lung cancer A549 cell clonogenic potential.

Ionizing radiation not only affects irradiated but also non-irradiated surrounding cells through intercellular communication, indicating that the former cells could affect the latter. The present study investigated the effect of X-irradiated normal human lung fibroblast WI-38 cells on the clonofenic potential of human lung cancer A549 cells by co-culturing them. Moreover, the relationship between the effects of co-culturing on the clonogenic potential of A549 cells and cellular senescence in WI-38 cells was investigated. The co-culture with 10-Gy-irradiated WI-38 cells and A549 cells enhanced the clonogenic potential of non- or X-irradiated A549 cells. Irradiated WI-38 cells exhibited high SA-β-gal activity, a cellular senescence hallmark. Importantly, treatment with senolytic drugs, which eliminate senescent cells, not only influenced high-SA-β-gal-activity cell percentages among the irradiated WI-38 cells but also the effect of irradiated WI-38 cells on the clonogenic potential of A549 cells. In conclusion, our results suggest that irradiated WI-38 cells promote A549 cell clonogenic potential and irradiated senescent WI-38 cells contribute to this effect.

Design, in silico studies and biological evaluation of novel chalcones tethered triazolo[3,4-a]isoquinoline as EGFR inhibitors targeting resistance in non-small cell lung cancer

A novel series of six [1,2,4]triazolo[3,4-a]isoquinolin-3-yl)-3-(1,3-diphenyl-1H-pyrazol-4-yl)prop-2-en-1-ones (3a–3f) was designed and synthesized. They were characterized based on spectral and elemental analyses. In silico studies were also committed to provide insights and a better understanding of their structural features. The six compounds were screened for their antiproliferative activity using the MTT assay against five human cancer cell lines, namely, A549, HCT116, PC3, HT29, and MCF-7 in parallel with the non-cancerous human lung cell line WI-38. The results showed that 3e and 3f have potential cytotoxic activities, especially on A549 cells with IC50 = 2.3 µM and 1.15 µM, respectively. Meanwhile, they recorded a minimal cytotoxic effect on WI-38 cells. Concerning the molecular mechanism of action, the present study showed the inhibitory effect of the six compounds against total EGFR. The most potent EGFR inhibitors were 3e and 3f with IC50 = 0.031 µM and 0.023 µM, respectively. The selectivity index of 3f for EGFRT790M was 1.81 times more selective than that of lapatinib. In addition, 3e and 3f initiated cell cycle arrest at the G2/M and pre-G1 phases along with the downregulation of anti-apoptotic protein Bcl2 and the upregulation of pro-apoptotic proteins: p53, Bax, and caspases 3, 8, and 9. Further studies are recommended to evaluate animal models’ promising anticancer activity and molecular mechanism of triazolo[3,4-a]isoquinoline derivatives 3e and 3f.

Design, characterization, and DFT exploration of new quinazoline-N-substituted analogs: Anti-cancer activity and molecular docking insights

In this study, we report the synthesis of a new series of quinazoline-N-4-fluorophenyl derivatives 4a-d and their antitumor activity against human cancer cells. The derivatives were confirmed by NMR (1H & 13C) and LC–MS. Their cytotoxicity was assessed using the MTT assay on WI38, HCT116, and HepG2 cell lines. Compound 4b showed significant cytotoxicity with IC50 values of 7.34 ± 0.7 µM and 4.36 ± 0.3 µM against HCT116 and HepG2 cells, respectively, while compound 4a exhibited moderate activity against WI38 cells (IC50 = 31.86 ± 2.4 µM). Molecular docking studies indicated that all four compounds have good binding affinity toward CAIX (−37.40 kcal/mol).

Activated single nucleotide mismatch determination through toehold-embedded hairpin-mediated strand displacement reaction alongside CRISPR-Cas12a for detection of TP53 point mutation

The CRISPR-Cas12a-based diagnostic platform is renowned for its high sensitivity and, when coupled with the toehold-mediated strand displacement (TMSD) reaction, enables the detection of single nucleotide mismatch (SNM). However, conventional TMSD with the CRISPR-Cas12a system suffers from signal leakage due to overlapping activator sequences and low displacement efficiency. Here, we propose a toehold-embedded hairpin-mediated strand displacement (THMSD) method and leverage the magnesium attenuation effect of CRISPR-Cas12a for selective single-point mutation detection in the TP53 gene. This approach effectively mitigates signal leakage as the hairpin DNA retains partial crRNA targeting sequences within its loop, preventing sequence overlap with the trigger and direct Cas12a activation. Additionally, the magnesium attenuation effect reduces background signal and enhances selectivity. We investigated toehold length and mismatch position to detect SNM with a high discrimination factor (DF). This high DF allows the THMSD/CRISPR-Cas12a system to differentiate between TP53 R273H mutant and TP53 wildtype genes for cancer screening at low abundances, down to 0.1 % with a concentration of 20 pM. When used for screening between H1975 and WI38 cells, the statistically significant difference was attainable with polymerase chain reaction (PCR) template loading as low as 4 ng. These results pave the way for developing CRISPR-Cas12a-based methods to effectively detect TP53 mutations as lung cancer biomarkers.

Therapeutic Effects of Crocin Nanoparticles Alone or in Combination with Doxorubicin against Hepatocellular Carcinoma In vitro.

Crocin (CRO), the primary antioxidant in saffron, is known for its anticancer properties. However, its effectiveness in topical therapy is limited due to low bioavailability, poor absorption, and low physicochemical stability. This study aimed to prepare crocin nanoparticles (CRO-NPs) to enhance their pharmaceutical efficacy and evaluate the synergistic effects of Cro-NPs with doxorubicin (DOX) chemotherapy on two cell lines: human hepatocellular carcinoma cells (HepG2) and non-cancerous cells (WI38). CRO-NPs were prepared using the emulsion diffusion technique and characterized by transmission electron microscopy (TEM), scanning electron microscopy (SEM), Zeta potential, and Fourier transform infrared spectroscopy (FT-IR). Cell proliferation inhibition was assessed using the MTT assay for DOX, CRO, CRO-NPs, and DOX+CRO-NPs. Apoptosis and cell cycle were evaluated by flow cytometry, and changes in the expression of apoptotic gene (P53) and autophagic genes (ATG5 & LC3) were analyzed using real-time polymerase chain reaction. TEM and SEM revealed that CRO-NPs exhibited a relatively spherical shape with an average size of 9.3 nm, and zeta potential analysis indicated better stability of CRO-NPs compared to native CRO. Significantly higher antitumor effects of CRO-NPs were observed against HepG2 cells (IC50 = 1.1 mg/ml and 0.57 mg/ml) compared to native CRO (IC50 = 6.1 mg/ml and 3.2 mg/ml) after 24 and 48 hours, respectively. Annexin-V assay on HepG2 cells indicated increased apoptotic rates across all treatments, with the highest percentage observed in CRO-NPs, accompanied by cell cycle arrest at the G2/M phase. Furthermore, gene expression analysis showed upregulation of P53, ATG5, and LC3 genes in DOX/CRO-NPs co-treatment compared to individual treatments. In contrast, WI38 cells exhibited greater sensitivity to DOX toxicity but showed no adverse response to CRONPs. Although more in vivo studies in animal models are required to corroborate these results, our findings suggest that CRO-NPs can be a potential new anticancer agent for hepatocellular carcinoma. Moreover, they have a synergistic effect with DOX against HepG2 cells and mitigate the toxicity of DOX on normal WI38 cells.

Evaluation of the functional activity and specificity of the <i>hSLPI</i> gene promoter

BACKGROUND: One of the main problems of modern oncotherapy is the lack of selectivity of the antitumor drugs, leading to their systemic toxicity on the body. Targeted expression of therapeutic genes represents a new approach in cancer gene therapy. One of the ways to achieve the selectivity of action of the therapeutic genes towards tumor cells is the use of promoters that are active only in tumor cells, but not in normal cells. AIM: To evaluate the functional activity and specificity of the promoter of the human secretory leukocyte protease inhibitor hSLPI. MATERIALS AND METHODS: The promoter of the hSLPI gene was cloned into the promotorless vector pTurboGFP-PRL. The functional activity and specificity of the promoter were assessed by the expression of the mRNA of the TurboGFP reporter gene and the intensity of its luminescence in A549, MCF-7, HeLa tumor cells and WI-38 normal cells using real-time polymerase chain reaction with reverse transcription and fluorescence analysis, respectively. RESULTS: Despite the literature data, the promoter of the hSLPIa gene demonstrated high transcriptional activity only against HeLa cervical cancer tumor cells. At the same time, in the cells of lung adenocarcinoma A549 and breast cancer MCF-7, as in normal WI-38 cells, a reduced level of promoter activity was observed. CONCLUSION: The data obtained confirm the need for a preliminary assessment of the functional activity of tumor-specific promoters in relation to tumor and normal cells.

Downregulation of IGFBP7 Alleviates LPS-induced Inflammation and Apoptosis in WI-38 Cells via Enhancing Mitophagy.

Pediatric pneumonia is an inflammatory disease with a very high incidence. IGF binding protein 7 (IGFBP7) plays an important role in inflammatory diseases. However, the role of IGFBP7 in pediatric pneumonia and its mechanism have not been reported. Human embryonic lung (WI-38) cells were induced by lipopolysaccharide (LPS) to construct the cell inflammatory injury model. Subsequently, the expression of IGFBP7 was detected by qPCR and western blot. Next, IGFBP7 interference plasmid was constructed, and cell viability and apoptosis were detected by CCK8, flow cytometry and western blot. ELISA and other techniques were used to detect the inflammatory level. Autophagy and mitochondrial activities were detected by immunofluorescence and other techniques, and mitophagy-related proteins were detected by western blot. To further investigate the regulatory mechanism of IGFBP7, we administered cyclosporin A, a mitophagy inhibitor, and then detected apoptosis and inflammation. The expression of IGFBP7 was significantly increased in LPS-induced WI-38 cells. Interference with IGFBP7 expression in LPS-induced cells significantly increased cell activity, decreased apoptosis and cellular inflammation levels. During this process, mitophagy was enhanced. Further addition of cyclosporin A significantly reversed the protective effect of IGFBP7 knockdown. To be concluded, inhibition of IGFBP7 alleviates LPS-induced inflammation and apoptosis in WI-38 cells via enhancing mitophagy.

Antileishmanial, antitrypanosomal and anti-coronavirus activities of benzophenanthridine alkaloids and other specialized metabolites isolated from the root bark of Zanthoxylum zanthoxyloides (Lam.) B.Zepernick & Timler

Strong antileishmanial and antitrypanosomal activities were highlighted for the crude methanolic extract (IC50 = 0.61 and 2.15 μg/mL, respectively) of Zanthoxylum zanthoxyloides (Lam.) B.Zepernick & Timler root bark, as well as for its apolar partitions (cyclohexane: IC50 = 0.66 and 5.17 μg/mL, respectively and dichloromethane: IC50 = 0.07 and 0.22 μg/mL, respectively), with a good selectivity index (SI) towards WI-38 cells. In addition, cyclohexane and dichloromethane extracts exhibited a dose-dependent inhibition of human coronavirus HCoV-229E infection in hepatoma Huh-7 cells expressing or not the cellular protease TMPRSS2 (IC50 values of 5.29 μg/mL and 4.87 μg/mL, respectively). Fractionation of these active extracts led to the isolation of a new racemic benzophenanthridine alkaloid named zanthoxyloithrine (1), together with 13 known compounds. Their structures were elucidated by spectroscopic techniques including IR, UV, HR-MS, 1D and 2D NMR and electronic circular dichroism. In parallel, HR-ESI-MS/MS based dereplication and molecular networking analysis were performed to identify unpurified compounds in cyclohexane and dichloromethane extracts. Zanthoxyloithrine (1) showed strong antileishmanial (IC50 = 0.14 μM, SI = 52.0) and antitrypanosomal (IC50 = 0.36 μM, SI = 20.8) activities. In addition, compound (1) demonstrated a high antiviral activity against HCoV-229E with IC50 value of 6.70 μM in presence of TMPRRS2 and without significant toxicity on Huh-7 cells. Other purified benzo[c]phenanthridine alkaloids also showed anti-coronavirus and antiparasitic activities.

Ultrasonic synthesis, characterization, DFT and molecular docking of a biocompatible Zn-based MOF as a potential antimicrobial, anti-inflammatory and antitumor agent

Zinc metal–organic frameworks have emerged as promising candidates, demonstrating excellent biological properties stemming from the unique characteristics of MOFs and zinc. In this study, we employed a facile method to synthesize a zinc metal–organic framework [Zn(IP)(H2O)] using ultrasound irradiation, with the linker being isophthalic acid (IPA) (1,3-benzene dicarboxylic acid). The parent Zn-MOF and two Ag/Zn-MOF samples prepared via loading and encapsulation methods were comprehensively characterized using various techniques, including FT-IR, XRD, SEM, TEM, N2 adsorption–desorption isotherm, UV–vis spectroscopy and TGA. The parent Zn-MOF and two Ag/Zn-MOF samples exhibited a broad spectrum of antibacterial effects. Remarkably, genomic DNA of P. aeruginosa was effectively degraded by Zn-MOF, further supporting its potent antibacterial results. The free radical inhibition assay demonstrated a 71.0% inhibition under the influence of Zn-MOF. In vitro cytotoxicity activity of Zn-MOF against HepG-2 and Caco-2 cell lines revealed differential cytotoxic effects, with higher cytotoxicity against Caco-2 as explored from the IC50 values. This cytotoxicity was supported by the high binding affinity of Zn-MOF to CT-DNA. Importantly, the non-toxic property of Zn-MOF was confirmed through its lack of cytotoxic effects against normal lung cell (Wi-38). The anti-inflammatory treatment of Zn-MOF achieved 75.0% efficiency relative to the standard Ibuprofen drug. DFT and docking provided insights into the geometric stability of Zn-MOF and its interaction with active amino acids within selected proteins associated with the investigated diseases. Finally, the synthesized Zn-MOF shows promise for applications in cancer treatment, chemoprevention, and particularly antibacterial purposes.

Lentil Extract-Mediated Ag QD Synthesis: Predilection for Albumin Protein Interaction, Antibacterial Activity, and Its Cytotoxicity for Wi-38 and PC-3 Cell Lines.

Recent focus has been directed toward semiconductor nanocrystals owing to their unique physicochemical properties. Nevertheless, the synthesis and characterization of quantum dots (QDs) pose considerable challenges, limiting our understanding of their interactions within a biological environment. This research offers valuable insights into the environmentally friendly production of silver quantum dots (Ag QDs) using lentil extract and clarifies their distinct physicochemical characteristics, previously unexplored to our knowledge. These findings pave the path for potential practical applications. The investigation of the phytochemical-assisted Ag QDs' affinity for BSA demonstrated modest interactions, as shown by the enthalpy and entropy changes as well as the associated Gibbs free energy during their association. Steady-state and time-resolved fluorescence spectroscopy further demonstrated a transient effect involving dynamic quenching, predominantly driven by Forster resonance energy transfer. Additionally, the study highlights the potential broad-spectrum antibacterial activity of Ag QDs (<5 nm, a zeta potential of -3.04 mV), exhibiting a remarkable MIC value of 1 μg/mL against Gram-negative bacteria (E. coli) and 1.65 μg/mL against Gram-positive bacteria (S. aureus). They can readily enter cells and tissues due to their minuscule size and the right chemical environment. They cause intracellular pathway disruption, which leads to cell death. This outcome emphasizes the distinctive biocompatibility of the green-synthesized Ag QDs, which has been confirmed by their MTT assay-based cytotoxicity against the PC-3 and Wi-38 cell lines.

Synthesis, Photophysical Properties and Cytotoxic Activities of Isoxazole‐Containing Difluoroboron Complexes

ABSTRACTA large series of previously unknown isoxazole containing difluoroboron β‐diketonate has been designed and synthesized from the available starting compounds. The photophysical properties of obtained compounds were studied, and the effects of the substituents in aromatic and isoxazole cycles of the BF2 complexes on the fluorescence were investigated. The effect of solvatochromism were demonstrated. Cytotoxic activity against MCF‐7 (breast carcinoma), HCT‐116 (colon carcinoma), A549 (pulmonary carcinoma) and WI38 (fibroblasts from lung tissue) cell lines was tested and moderate toxicity in the concentration range of 10–100 μM was found for several compounds.

The green approach of chitosan/Fe2O3/ZnO-nanocomposite synthesis with an evaluation of its biological activities

Biopolymers embedded with nanoparticles of metal oxides (MOs) demonstrate a wide range of bio-functions. Chitosan-incorporated MOs are an interesting class of support matrices for enhancing the biological function, compared to other support matrices. Therefore, the importance of this study lies in exploiting chitosan as a carrier not of one metal as in previous studies, but of two metals in the form of a nanocomposite to carry out several biological functions. The coprecipitation approach was employed to synthesize chitosan/Fe2O3/ZnO-nanocomposite in the present research. The characterization of chitosan/Fe2O3/ZnO-nanocomposite was performed to find out the morphology and dispersion properties of chitosan/Fe2O3/ZnO-nanocomposite. The X-ray diffraction (XRD) investigation revealed that these were crystalline. Fourier transforms infrared (FTIR) spectrum bands were viewed at 400/cm and 900/cm, due to the stretching vibration of Fe and Zn oxygen bond. TEM showed that chitosan/Fe2O3/ZnO-nanocomposite was of 20–95 nm in size. chitosan/Fe2O3/ZnO-nanocomposite exhibited inhibitory potential against Staphylococcus aureus, Bacillus subtilis, Escherichia coli, and Candida albicans with inhibition zones of 25 ± 0.1, 28 ± 0.2, 27 ± 0.1, and 27 ± 0.2 mm, respectively while didn’t inhibited Aspergillus niger. MIC value of nanocomposite was 15.62 ± 0.33 µg/mL for C. albicans, B. subtilis and E. coli, while it was 62.50 ± 0.66 µg/mL for Pseudomonas aeruginosa. Ranged values of nanocomposite MBC (15.62 ± 0.33 to 125 ± 1 µg/mL) were attributed to all tested bacteria. Different concentrations of chitosan/Fe2O3/ZnO-nanocomposite MBC (25, 50, and 75%) reflected anti-biofilm activity against E. coli (85.0, 93.2, and 96.0%), B. subtilis (84.88, 92.21, and 96.99%), S. aureus 81.64, 90.52, and 94.64%) and P. aurogenosa (90.11, 94.43, and 98.24%), respectively. The differences in the levels of antimicrobial activities may depend on the type of examined microbes. Antioxidant activity of chitosan/Fe2O3/ZnO-nanocomposite was recorded with excellent IC50 values of 16.06 and 32.6 µg/mL using DPPH and ABTS scavenging, respectively. Wound heal by chitosan/Fe2O3/ZnO-nanocomposite was achieved with 100% compared to the untreated cells (76.75% of wound closer). The cytotoxicity outcomes showed that the IC50 of the chitosan/Fe2O3/ZnO-nanocomposite was 564.32 ± 1.46 µg/mL normal WI-38 cells. Based on the achieved findings, the chitosan/Fe2O3/ZnO-nanocomposite is a very promising agent for perform pharmacological activities.

USP9X promotes LPS-induced fibroblast cell apoptosis, inflammation and oxidative stress by regulation of TBL1XR1 deubiquitination

Abstract Background Ubiquitination and deubiquitination are involved in the progression of human diseases, including acute pneumonia. In this study, we aimed to explore the functions of ubiquitin-specific peptidase 9X-linked (USP9X) in lipopolysaccharide (LPS)-treated WI-38 cells. Methods WI-38 cells were treated with LPS to induce the cellular damage and inflammation. 3-(4, 5-dimethyl-2-thiazolyl)-2, 5-diphenyl-2-H-tetrazolium bromide (MTT) assay and 5-ethynyl-2’-deoxyuridine (EdU) assay were performed to examine the proliferation of LPS-treated WI-38 cells. Flow cytometry analysis was conducted to detect LPS-treated WI-38 cell apoptosis. ELISA kits were utilized to determine the concentrations of inflammatory factors (IL-1β and TNF-α). Superoxide dismutase (SOD) activity and reactive oxygen species (ROS) level were examined with related kits. Ubibrowser (http://ubibrowser.bio-it.cn/ubibrowser/), ubiquitination assay and Co-Immunoprecipitation (Co-IP) assay demonstrated the interaction between USP9X and transducin β-like 1X related protein 1 (TBL1XR1). qRT-PCR assay and western blot assay were manipulated to determine the expression of USP9X and TBL1XR1. TBL1XR1 and USP9X knockdown experiments were conducted to explore their functions on LPS-induced WI-38 cell injury and inflammation. Results TBL1XR1 expression was upregulated in LPS-treated WI-38 cells. TBL1XR1 knockdown promoted cell proliferation and repressed apoptosis, inflammation and oxidative stress in LPS-treated WI-38 cells. Moreover, USP9X deubiquitinated TBL1XR1 to regulate TBL1XR1 expression. USP9X knockdown restored the effects of LPS on WI-38 cell proliferation, apoptosis, inflammation and oxidative stress, but these effects of USP9X knockdown were further abolished by TBL1XR1 overexpression. Additionally, USP9X promoted the NF-κB signaling pathway by the deubiquitination of TBL1XR1. Conclusion USP9X promoted the apoptosis, inflammation and oxidative stress of LPS-stimulated WI-38 cells through the deubiquitination of TBL1XR1.

Synthesis and computational studies of new pyridine, pyrazole, pyran, and pyranopyrimidine-based derivatives of potential antimicrobial activity as DNA gyrase and topoisomerase IV inhibitors

In this study, new derivatives based on pyridine, pyrazole, pyran, and pyranopyrimidine scaffolds 5–14 were designed and synthesized. The newly synthesized compounds were characterized using microanalytical and spectral data. Using amoxicillin and amphotericin B, two common antibacterial and antifungal medications, respectively, the antimicrobial activity of the new compounds was assessed against a variety of strains of both Gram-positive and Gram-negative bacteria as well as fungal infections. The most promising activity was achieved by pyridine-based derivatives 5, 7a, and 7b, which exhibited MIC values ranging from 0.95 to 18.04 µM against the examined bacterial strains and ranging from 4.65 to 33.16 µM against the examined fungal strains. The later derivatives were also evaluated as E. coli DNA gyrase B/topoisomerase IV inhibitors in comparison with novobiocin as a standard drug. Although the assessed candidates 5, 7a, and 7b exhibited a promising suppression impact against E. coli DNA gyrase B (IC50 = 3.11 ± 0.14, 3.85 ± 0.20, and 4.05 ± 0.10, respectively, IC50 novobiocin= 3.64 ± 0.10 µM), they showed detectable less suppression impact against Topoisomerase IV than that of the reference drug with IC50 values of 19.72 ± 1.00, 79.33 ± 0.25, 85.14 ± 0.10, and 13.42 ± 0.05 µM, respectively. Additionally, the safety profiles of 5, 7a, and 7b against WI38 cells were significantly superior to that of novobiocin (IC50 = 138 ± 0.33, 170 ± 0.40, 163.3 ± 0.17, and 86.2 ± 0.03 µM, respectively).Lastly, molecular docking was used to examine the most active derivatives 5, 7a, and 7b's binding capacity to the target DNA gyrase B/topoisomerase IV. The molecules exhibited various H-interactions with the amino acid residues Asp73, Arg76, and Val71 in E. coli DNA gyrase, where compound 5 exhibited H-bonding with the Asp1069 side chain of E. coli Topoisomerase IV. Additional in silico ADMET studies were conducted to represent their oral bioavailability and drug-likeness characteristics.