Research in Pharmaceutical Sciences最新文献

Background and purpose: In initiating and regulating immune responses, dendritic cells (DCs) play an important role as antigen-presenting cells. When DCs are exposed to tumor cell lysates, they can stimulate T cells to recognize tumor-associated and tumor-specific antigens and generate an immune response against cancer. The purpose of this study was to compare 4 different approaches for preparing breast tumor cell lysates for pulsing DCs.

Experimental approach: To prepare tumor cell lysates from 4T1 cells, 4 different methods were used, including freeze-thaw, hypochlorous acid (HOCl), hyperthermia, and ultraviolet-B irradiation. The effects of the tumor lysates were assessed on the maturation of DCs and the secretion of cytokines using flow cytometry and ELISA. Furthermore, DCs pulsed with different lysates were also evaluated for their ability to promote CD8+ T cell proliferation and release cytokines.

Findings/results: The results demonstrated that DCs pulsed with lysate prepared by HOCl exhibited more maturation surface biomarker expression (CD86) than DCs pulsed with freeze-thawed cells or unloaded DCs (control). Furthermore, activated DCs were also found to promote CD8+ T cell proliferation and induce the responses of T cells by producing high levels of IFN-γ, while inhibiting IL-10.

Conclusion and implications: HOCl is capable of releasing tumor antigens while maintaining its ability to stimulate an immune response. DCs-based therapies may be designed based on the findings presented here, demonstrating a cross-presentation of antigens and specific activation of the immune system against breast cancer.

Background and purpose: T-cell immunoglobulin and mucin-domain containing protein-3 (TIM-3)/ galectin-9 (Gal-9)/ autocrine loop in myeloid leukemia stem cells provokes inflammation through the NF-κB signaling pathway, which is influential in the expression of inflammatory factors. Interleukin 1β (IL-1β) is a vital inflammatory cytokine that plays an important role in the proliferation and therapy resistance of acute myeloid leukemia (AML) cells. This study aimed to assess the effect of Gal-9 on IL-1β in the human leukemic U937 cell line.

Experimental approach: The U937 cells were cultured in different concentrations of Gal-9. Cell counting kit-8 was used to assess the effect of Gal-9 on human leukemic U937 cell proliferation. Also, its impact on the expression of TIM-3, Gal-9, IL-1β, IL-1βR, IL-1βRAP, and NLRP3 genes and IL-1β protein was studied by RT-PCR and ELISA, respectively. Moreover, the effect of Gal-9 on the NF-κB signaling pathway was evaluated by western blotting.

Findings/results: U937 cells were expanded in the presence of Gal-9 in a concentration-dependent manner. Following treatment of U937 cells with Gal-9, the gene expression of Gal-9, IL-1B, IL-1BR, and IL-1BRAP were significantly upregulated compared to the control group. The IL-1β concentration increased following Gal-9 treatment in a concentration-dependent manner, while following time-pass its level significantly decreased. Furthermore, Gal-9 slightly increased NF-κB phosphorylation.

Conclusion and implications: Gal-9 increased IL-1β level as a critical inflammatory cytokine in the proliferation and resistance of AML cells to therapy. According to this finding, targeting and blocking the TIM-3/Gal-9 autocrine loop can suppress IL-1β production and facilitate AML treatment.

Background and purpose: The study aimed to design new chimeric antimicrobial peptides targeting Acinetobacter baumannii, a widespread and growing global concern due to antibiotic resistance. Three bioinformatics-based methods were utilized for this purpose.

Experimental approach: To design new chimeric peptides targeting Acinetobacter baumannii, a group of peptides were initially selected and divided into two categories based on their scores and performance. The peptides were then combined through 3 methods: 1. combining sequences based on their secondary structure using GOR IV software; 2. grouping only the amino acid sequences involved in the formation of the target peptide helix structure using Accelrys DS visualizer software; and 3. combining the most similar parts of the peptides in terms of amino acid type and order using online AntiBP2 software. The sequence length was optimized, and some amino acids were substituted.

Findings/results: The M-CIT peptide was selected for synthesis in the first method, but it did not show significant activity against the target bacteria (MIC = 187.5 μM and MBC = 375 μM). In the second method, no suitable score was observed. However, the M-PEX12 peptide was synthesized in the second method, demonstrating antimicrobial activity against A. baumannii (MIC = 33.1 μM and MBC = 41.4 μM).

Conclusion and implications: Three methods were evaluated for designing new chimeric peptides, and the third method, which involved modifying the number of amino acids in the parental peptides while maintaining their similarity, was found to be the most suitable.

Background and purpose: Melatonin is a product of the pineal gland, which regulates the circadian cycle. Neurotoxicity is the most important side effect of methamphetamine (Met) abuse during pregnancy. This study aimed to explore the effect of Met exposure during gestation and lactation periods on the learning and memory of offspring mice. The protective effect of melatonin and the role of oxidative stress and acetylcholinesterase were also investigated.

Experimental approach: The pregnant mice were randomly divided into 2 groups. Saline or Met (5 mg/kg) was injected daily during pregnancy and lactation. After the lactation period, the offspring mice of each group were divided into 2 subgroups, and saline or melatonin (10 mg/kg) was orally (gavage) administered to the offspring mice from the post-delivery (PD) day 21 up to PD Day 60. The offspring mice were examined in the passive avoidance (PA) test. Finally, oxidative stress markers and acetylcholinesterase (AchE) activity were measured in the brains.

Findings/results: As a result, Met decreased delay and light time while increasing the frequency of entry and time in the dark region of PA. However, melatonin alleviated the impairing effect of Met on PA performance. Meanwhile, the administration of Met increased malondialdehyde while decreasing superoxide dismutase and thiol content. Furthermore, AchE activity was significantly increased in Met-treated mice. Melatonin reversed the levels of antioxidants, lipid peroxidation, and AchE activity in the brain.

Conclusion and implications: Together, these results suggested that melatonin may be a potential therapeutic agent for alleviating Met-induced memory impairment by restoring redox hemostasis and AchE.

Background and purpose: Snake venom contains numerous proteolytic enzymes, including metalloproteases. This study aimed to isolate, characterize the metalloproteinase from Montivipera raddei venom and insert its cDNA into a host cell genome for expressing the protein's potential use as a coagulation factor.

Experimental approach: Initially, M. raddei venom was analyzed using SDS-PAGE and ion-exchange chromatography. All purified fractions were assessed using a prothrombin time (PT) assay. Immunoblot analysis and MALDI-TOF/TOF MS spectrometry confirmed the metalloproteinase active domain in the obtained fraction. All of the confirmations were studied using the I-TASSER server. To produce the recombinant enzyme, metalloproteinase cDNA was isolated from the total RNA of the Caucasus viper venom gland tissue and cloned into pEX-A2-H plasmid, containing homologous sequences with CHO-S cell genome. Also, it was confirmed by western blotting and PT test.

Findings/results: Two fractions from M. raddei venom had PT values of 6 and 21 s, respectively. We confirmed the presence of a 25-KDa procoagulant metalloprotease (Met). The results indicated successful expression of the Met protein in the recombinant CHO-S cells. The Met protein was structurally similar to the target in the PDB ID. 2e3x, regulatory subunit of the blood coagulation factor X- and IX-activating enzyme. The Met protein also contained a similar binding domain with 60 amino acids, comparable to those in E. carinatus. and E. pyramidum leakeyi metalloproteinases.

Conclusion and implications: The Met protein may be a potent candidate recombinant coagulant drug. Remarkably, the thrombin time for this protein was 3 s.

Despite great advances in cancer identification and treatment, malignancies remain the primary cause of high morbidity and mortality worldwide. The drawbacks of conventional chemotherapy, such as severe toxicity, lack of specificity related to actively dividing cells, and resistance, can warrant the urgent need to develop an alternative approach to treat this disease. To overcome the drawbacks, researchers are attempting to deliver drugs to the site of action (targeted delivery) or to identify drugs that specifically target tumor cells. In this regard, highly cationic and amphipathic antimicrobial peptides are attracting the attention of researchers due to their potent anticancer activity, low cost of manufacture, and, most critically, tumor-targeting activity. A growing number of documents have shown that some of the mentioned peptides exhibited a broad spectrum of cytotoxic activity against cancer cells but not normal mammalian cells entitled as anticancer peptides. Due to their solubility, low toxicity, strong tumor penetration, high selectivity, and ability to be used alone or in conjunction with other conventional medications, anticancer peptides have the potential to become very successful cancer treatments in the future. This review provided an overview of the studies concerning anticancer peptide classification, modes of action, and selectivity, and also summarized some of the anticancer peptides developed for targeting different types of malignancies. The role of in silico methods or artificial intelligence in the design and discovery of anticancer peptides was briefly explained. Additionally, the current review addressed challenges in utilizing anticancer peptides for cancer therapy and highlighted peptides currently undergoing clinical trials.

Background and purpose: Padina pavonica, a brown alga, displays protection against oxidative stress, neuroinflammation, and neurodegenerative disorders. Considering the beneficial effects of P. pavonica and since its antidepressant effects have not yet been studied, we investigated its methanol and hexane partitions (PMP and PHP) in mice model.

Experimental approach: In male mice (25 ± 2 g), depression was initiated by administrating dexamethasone (15 μg/kg) subcutaneously. PMP or PHP (80-160 mg/kg) was administered intraperitoneally. All the injections continued for two weeks. After the locomotor test, different depression criteria were evaluated by forced swim test (FST), marble burying test (MBT), sucrose preference (SP) test, and novelty-suppressed feeding test (NSFT).

Findings/results: PMP (160 mg/kg) showed antidepressant effects, immobility time decreased significantly during FST (84.6 ± 10.4 s) compared to the control, food intake increased significantly during NSFT (21.1 ± 2.5 mg/g) versus the control, and SP was 81%. PHP (80 mg/kg) reduced immobility time to 124.5 ± 6.7 s compared to the vehicle and increased the SP to 85%. Dexamethasone-induced depression. While co-treatment with PMP or PHP prevented depression initiated by dexamethasone. Only PHP reduced the number of buried marbles after 30 min against the control. The applied doses did not cause significant changes in the locomotor activity.

Conclusion and implications: PMP and PHP exhibited antidepressant-like effects in mice. PHP also reduced the number of buried marbles, implying a reduction in obsessive-like behavior. These observations underscore the constituents inherent in PMP and PHP, which merit further exploration to elucidate their potential therapeutic applications.

Background and purpose: Type 2 diabetes mellitus (T2DM) is a metabolic disorder characterized by β-cell dysfunction, insulin resistance, and elevated blood sugar levels. Several studies have explored the therapeutic potential of coenzyme Q₁₀ (CoQ₁₀) in managing diabetes, but no reports have examined the possible mechanism of CoQ₁₀ in T2DM. Here, we reported that CoQ₁₀ protects pancreatic β-cell structure and function by modulating the expression of mir-33a/mir-21/SREBP1 and described more detailed tissue alterations.

Experimental approach: The study randomly divided rats into three groups (n = 10): control, diabetic, and diabetic + CoQ₁₀. The diabetic + CoQ₁₀ group consisted of diabetic rats that were concurrently administered CoQ₁₀ (20 mg/kg/i.p.) three days/week for eight weeks. In addition to microscopic examination, the study involved evaluating glucose, insulin, and oxidative profiles in the serum and analyzing the levels of cholesterol, mir-33a, mir-2i, and SREBP1 in pancreatic tissue.

Findings/results: Our results revealed that CoQ₁₀ restores glucose/insulin homeostasis, oxidative parameters, cholesterol levels, and the expressions of mir-33a, mir-21, and SREBP1. In addition, the CoQ₁₀-treated diabetic rats showed increased active β-cells compared to the diabetic group. The immunohistochemical examination of insulin revealed a higher quantity and larger size of pancreatic islets in the experimental group.

Conclusion and implications: The restoration of β-cell integrity following treatment with CoQ₁₀ may elucidate the therapeutic benefits of this compound in diabetes management, potentially through its influence on the pancreatic expression of mir-33a/mir-21/SREBP1, subsequently maintaining healthy tissue.

Background and purpose: Ha-rak (HR), an equal-proportion combination of roots from Capparis micracantha DC., Clerodendrum petasites S. Moore., Ficus racemosa L., Harrisonia perforata (Blanco) Merr., and Tiliacora triandra (Colebr.) Diels, Piper betle L. (PB), and Garcinia mangostana L. (GM) are commonly used in traditional Thai medicine to treat skin diseases, including atopic dermatitis (AD). Combining three medicines in adjusted proportions can improve efficacy, reduce toxicity, and reduce medication. This study aimed to evaluate the anti-allergic, anti-inflammatory, antimicrobial, and cytotoxic activities of the ethanolic extracts of different combinations to analyze the relationship among elements, medicinal tastes, and biological activities.

Experimental approach: The biological activities (anti-allergic, anti-inflammatory, and anti-microbial activities and cell viability) of ethanolic extracts of plants and their combinations in various proportions were evaluated, as well as the chemical content of the developed remedies using the HPLC technique.

Findings/results: HMB-123 was the most significantly effective combination for AD. HMB-123 reduced β-hexosaminidase release from RBL-2H3 cells to a greater extent than chlorpheniramine. HMB-123 significantly inhibited nitric oxide and TNF-α production in LPS-stimulated RAW 264.7 cells. HMB-123 demonstrated antibacterial activity against all tested bacteria and antifungal activity against Candida albicans. For single extract, PB exhibited the highest anti-fungal activity, while GM exhibited the highest anti-bacterial activity.

Conclusion and implications: The combined extracts showed potential as an optimized remedy for AD. HMB-123 demonstrated the highest anti-allergic, anti-inflammatory, and antimicrobial activities, making it a promising development candidate for AD treatment. To confirm safety and efficacy, further pre-clinical and clinical testing is necessary.

Background and purpose: Due to delivery obstacles, Simvastatin, a potential anticancer agent, faces clinical limitations. This study aimed to enhance simvastatin delivery and efficacy against triple-negative breast cancer (TNBC) by developing liposomes modified with sodium oleate (NaOL) to improve endosomal escape.

Experimental approach: Simvastatin was encapsulated in 1,2-dimyristoyl-sn-glycero-3-phosphocholine/cholesterol liposomes through thin film hydration. Liposomes with poly(lactic-co-glycolic acid) (PLGA), individually modified with NaOL and PLGA, served as a control endosomal escape enhancer. Formulations were characterized for size, charge, and encapsulation efficiency. Endosomal escape was quantified through subcellular colocalization analysis using confocal microscopy, and anticancer activity was assessed by evaluating cytotoxicity against 4T1 TNBC cells, followed by measurements of intracellular reactive oxygen species (ROS) and DNA damage.

Findings/results: Unmodified liposomes had a size of 115.2 ± 7.94 nm, a zeta potential of -9.67 ± 3.01 mV, and an encapsulation efficiency of 78.93% ± 6.72. NaOL-modified liposomes had a size of 119 ± 9.37 nm, a zeta potential of -31.05 ± 2.38 mV, and an encapsulation efficiency of 84.96% ± 2.51. While PLGA-modified liposomes had a size of 151.1 ± 7.35 nm, zeta potential of -18.68 ± 1.41 mV, and encapsulation efficiency of 83.63% ± 5.56. Importantly, NaOL-liposomes exhibited lower IC₅₀ values, improved endosomal escape, and enhanced anticancer activity compared to unmodified liposomes.

Conclusion and implications: Surface modification with NaOL is a promising strategy to enhance the anticancer efficacy of simvastatin liposomes against TNBC through improved endosomal escape. These encouraging in-vitro findings warrant further in-vivo investigations into the potential for NaOL-modified liposomes to improve TNBC patient outcomes.