Advances in Pharmacological and Pharmaceutical Sciences最新文献

Chagas disease caused by the obligate intracellular flagellate protozoan Trypanozoma cruzi infects about 6 million people. From the 1930s to the present, the antitumor capacity of T. cruzi has been studied; however, the identification of the responsible molecules for this effect remains undiscovered. Calcineurin, a calcium/calmodulin-dependent serine/threonine phosphatase, is a heterodimer consisting of a catalytic subunit (CaNA) and a regulatory subunit (CaNB). It has been described that T. cruzi CaN is involved in the cell invasion and proliferation of the parasite. Recently, extracellular human CaNB has been demonstrated to be capable of inhibiting tumor growth cells, conferring an antitumor effect; however, the extracellular role of T. cruzi CaNB (TcCaNB) is still unknown. The objective of this work was to investigate the antitumor potential of TcCaNB by interacting with membrane proteins and evaluating its effects on the viability, proliferation, and morphology of tumor cells in vitro. Additionally, the possible mechanism of action of TcCaNB was explored. Murine melanoma (B16-F10), human cervical adenocarcinoma (HeLa), and African green monkey kidney epithelial (Vero) cell lines were employed for in vitro assays. Far Western blot and immunofluorescence were performed to assess the interaction of TcCaNB with membrane proteins, and the effect of TcCaNB on cell viability and proliferation was evaluated using the MTS assay and the CyQUANT NF assay, respectively. The effect of the caspase inhibitor Z-VAD-FMK on TcCaNB-stimulated tumor cells was investigated to determine if TcCaNB-induced cell death was associated with apoptosis. To assess cell cycle progression, TcCaNB-treated cells were analyzed by flow cytometry. In this study, the results showed an interaction of TcCaNB with cell membrane proteins in B16-F10 and HeLa tumor lines, indicating that TcCaNB is capable of decreasing viability and proliferation of B16-F10 and HeLa cells, with no significant effect observed in Vero cells. Furthermore, morphological changes were observed in tumor cells treated with TcCaNB. DNA fragmentations and inhibition of caspases with Z-VAD-FMK partially counteracted the cytotoxic effects of TcCaNB on tumor cells, suggesting that TcCaNB-induced cell death might be associated with apoptosis. Additionally, TcCaNB caused S phase cell cycle arrest in HeLa cells, with an increase in the sub-G1 population indicative of apoptosis, while no significant effects were observed in Vero cells.

The discovery and characterization of the signal cascades of the β-adrenergic receptors have made it possible to effectively target the receptors for drug development. β-Adrenergic receptors are a class A rhodopsin type of G protein-coupled receptors (GPCRs) that are stimulated mainly by catecholamines and therefore mediate diverse effects of the parasympathetic nervous system in eliciting "fight or flight" type responses. They are detectable in several human tissues where they control a plethora of physiological processes and therefore contribute to the pathogenesis of several disease conditions. Given the relevance of the β-adrenergic receptor as a molecular target for many pathological conditions, this comprehensive review aims at providing an in-depth exploration of the recent advancements in β₃-adrenergic receptor research. More importantly, we delve into the prospects of the β₃-adrenergic receptor as a therapeutic target across a variety of clinical domains.

In traditional Congolese medicine, the plants Crassocephalum montuosum (CrasMon) and Crassocephalum picridifolium (CrasPic) are used to treat bacterial gastroenteritis. In the present study, the antibacterial and antioxidant activities as well as the acute and subacute toxicity of organic extracts from the whole plant of the two investigated taxa were evaluated. Physicochemical parameters were also determined, and total phenolics, flavonoids, and tannins were investigated and assayed. The antibacterial and antioxidant activities of the plant extracts were evaluated using disc diffusion, tube macrodilution, and DPPH tests. Conversely, traditional solution reactions, gravimetric tests, and spectrophotometric tests were used to generate physicochemical profiles, identify secondary metabolite groups, and perform microdilution and DPPH tests to evaluate the antibacterial and antioxidant activities, respectively. OECD tests were adapted to assess the acute and subacute toxicity. All the extracts showed antibacterial activity against E. coli and S. typhi strains with the diameter zone of inhibition (DZI) ranging from 12 to 23 mm and the minimum inhibitory concentration (MIC): 15.625-125 μg·mL^-1. The methanolic extract of CrasPic showed the most pronounced activity with a DZI of 21-23 mm and MIC of 15.625-62.5 μg·mL^-1. All extracts showed high antioxidant activity with IC₅₀ (half maximal inhibitory concentration) ranging from 11.6 to 21.8 μg·mL^-1, with the methanolic extract of CrasMon showing the most pronounced activity. Both plants contain a variety of phytochemicals including coumarins, quinones, flavonoids, phenols, saponins, tannins, and terpenoids. The methanolic extract of CrasPic exhibits the highest content of total phenolics (300 mg·GAE·g^-1), flavonoids (56 mg·QE·g^-1), and tannins (155 mg·GAE·g^-1). These extracts have a median lethal dose (LD₅₀) > 5000 mg·kg^-1 and no signs of toxicity at 200 mg·kg^-1 after 30 days of oral administration to Cavia porcellus. The total ash content was determined to be 14.2% and 15.8% (on a dry weight basis), with the ash insoluble in hydrochloric acid exhibiting a range of 4.04%-5.03%. CrasMon and CrasPic have been demonstrated to exhibit a good antibacterial and antioxidant activities, at least in part, due to the presence of phenolic compounds. These activities may provide a rationale for their use in traditional Congolese medicine against gastroenteritis.

ER stress is a crucial factor in the progression of vascular cell diseases. Notably, octanoic acid (OA; C8:0) and decanoic acid (DA; C10:0), prominent components of medium-chain fatty acids (MCFAs), may provide potential health benefits. However, their effects on vascular smooth muscle cells (VSMCs) remain unknown. Given the link between ER stress and vascular cell pathological conditions, the primary goal of this research is to investigate the protective effects of OA and DA against ER stress induction in rat aortic smooth muscle cells (RASMCs). To achieve this objective, RASMCs were pretreated with OA and DA at concentrations of 250 and 500 μM for 24 h. Subsequently, the cells were exposed to 1 μg/mL of tunicamycin, an ER stress inducer, for an additional 24 h. Apoptosis was assessed using DAPI staining, while DCFH₂-DA probe was used to measure ROS levels. Furthermore, the gene expression of ER stress markers, such as CHOP, GRP78, ATF4, and eIF2α, as well as contractile markers like αSMA and MYH11, was assessed using real-time reverse transcription polymerase chain reaction. Moreover, the αSMA protein level was measured using immunocytochemistry techniques. The study revealed that OA and DA significantly mitigated cell death caused by tunicamycin, decreased ROS production, and inhibited the gene expression of ER stress markers (CHOP, GRP78, and eIF2α). Notably, OA and DA also inhibited the expression of contractile genes (α-SMA and MYH11) and reduced the number of α-SMA-positive cells in tunicamycin-treated RASMCs. These findings indicate that OA and DA offer protection against ER stress-stimulated cell death and ROS generation in VSMCs, thereby supporting their potential therapeutic applications for safeguarding these cells.

Hepatocellular carcinoma (HCC) is one of the most aggressive types of liver cancer, and it is frequently associated with upregulated c-Myc expression. Sorafenib (Sor) is commonly used to treat HCC, but many patients experienced mild to severe side effects due to prolonged Sor treatment during therapy. It has been known that Pentagamavunone-1 (PGV-1) exhibits a remarkable antiproliferative effect on several cancer cells, yet limited studies have reported its cellular activities in HCC. The current study aims to evaluate the anticancer effects of Sor in combination with PGV-1 on the progression of HCC proliferation. c-Myc expressing cells, JHH-7 and Huh-7, were used for this study, then Sor and PGV-1 were tested for their effect on the cellular physiology phenomena including cytotoxicity combination assay and colony formation assay, cell cycle profile and reactive oxygen species (ROS) level by flow cytometry, senescence induction by beta-galactosidase (SA-β-gal) assay, and migration inhibition by wound healing assay. The c-Myc expression was evaluated through Western blot. PGV-1 was more effective than Sor at inhibiting cell growth, and it showed greater selectivity for HCC over fibroblast cells. The combination of Sor with PGV-1 exhibited synergistic-additive cytotoxicity with an irreversible effect in HCC cell lines. The combination induced senescence similarly with Sor alone in JHH-7 cells, while PGV-1 enhanced the cellular senescence when combined with Sor in Huh-7 cells. Furthermore, the combination increased ROS level in the same way as PGV-1 did in HCC. The combination with PGV-1 acted better than Sor alone to inhibit JHH-7 cell migration. In addition, the combination treatment led to the suppression of c-Myc, particularly in JHH-7 cells. Taken together, combining Sor with PGV-1 promotes better efficacy than Sor alone to inhibit HCC cell proliferation, and further evaluation of the efficacy and safety of adding PGV-1 to Sor in HCC therapy is worthwhile as a potential combination treatment option.

High mortality and disability have been principally linked with traumatic brain injury (TBI) with this condition being a principal issue in public health. Currently, there is no reliable pharmaceutical agent to salvage the damage caused by TBI. Baicalein (BCL), a traditional Chinese medicine active ingredient, has preliminarily shown repair activity on brain injury. However, BCL has poor water solubility and bioavailability, which culminates in rapid elimination in vivo. Herein, we sought to solve the above-mentioned challenges that are associated with the application of this flavonoid by preparing BCL-loaded long-circulating liposomes (BLC-Lips) via thin-film hydration method. Subsequently, BLC-Lips were applied to TBI model mice to evaluate their effect on brain injury repair. The results showed that the prepared BLC-Lips exhibited smaller sized nanoparticles, excellent polydispersed index (PDI), and zeta (ζ)-potential with stable property. After loading of BCL into the liposomes, we estimated the physicochemical properties of BLC-Lips to be roughly 87.98% (encapsulation efficiency [EE]) and 7.56% (loading capacity of the drug). Administration of BLC-Lips through oral route increased aqueous solubility, bioavailability, and time for in vivo circulation of BCL. Moreover, the BLC-Lips could improve outcomes of histological and neurological motor function and reduce inflammatory cytokines and neurotoxicity after TBI. Taken together, the long-circulating liposomes may serve as a new approach to potentially prolong drug circulation in vivo and increased bioavailability of BCL for TBI treatment.

Obesity is a global health concern, steadily rising and posing risks to various health conditions. Despite available antiobesity drugs, their withdrawal due to severe side effects highlights the need for safer alternatives. Natural products, particularly mixed herbal formulations, present a promising avenue in obesity research. This study aimed to investigate the potential antiobesity effects of the NL herbal formula, a traditional remedy in Nakhon Si Thammarat, Thailand, composed of nine herbs. The specific focus was on the inhibitory effects on α-glucosidase and pancreatic lipase enzyme activities, adipogenesis inhibition and lipolysis promotion. NL extract was phytochemically analyzed and assessed for its inhibitory effects on α-glucosidase and pancreatic lipase. Its impact on lipid accumulation and glycerol release was also evaluated. Phytochemical analysis using liquid chromatography-tandem mass spectrometry (LC/MS-MS) identified piperine as the major compound in the NL extract. NL extract exhibited significant inhibition of α-glucosidase, moderate pancreatic lipase inhibition, and dose-dependent reduction in fat accumulation and triglyceride content. Glycerol release increased compared to the control, indicating potential benefits in weight management. This research underscores the potential of the NL formula in combating obesity through its effects on adipogenesis, lipolysis, and enzyme activities. Further investigations into the molecular mechanisms are warranted to fully elucidate its therapeutic potential.

Saikosaponin-b2 (SS-b2), an active ingredient derived from the root of Radix Bupleuri, possesses antitumour, anti-inflammatory, antioxidative and hepatoprotective properties. We investigated the inhibition of tumour proliferation by SS-b2 and the underlying molecular mechanisms, including the MACC1/p-c-Met/p-Akt pathway expression in HepG2 liver cancer cells and H22 tumour-bearing mice. Animal experiments showed that SS-b2 significantly decreased the levels of MACC1, p-c-MET and p-Akt in tumour tissue transplanted with H22 liver cancer cells in mice, while it increased the expression of p-BAD. The results also revealed a concentration-dependent suppression of MACC1, p-c-Met and p-Akt expression in the SS-b2 treatment group compared with the control group. Additionally, the suppression of MACC1 activation by SS-b2 resulted in a reduction in the viability and proliferation of HepG2 liver cancer cells, and this reduction was comparable to that by doxorubicin (DOX). This suggests that SS-b2 has significant efficacy in liver cancer, comparable to DOX. Meanwhile, Annexin V-FITC/PI staining and western blot analysis of cleaved caspase 9 and cleaved caspase 3 demonstrated that SS-b2 induced apoptosis of HepG2 liver cancer cells. These findings provide experimental evidence suggesting that SS-b2 is a promising anticancer agent for liver cancer.

Recent considerations of natural sources as potential anticancer agents have arisen due to the origins of numerous drugs commonly used in chemotherapy. Plant-based drugs, in particular, have attracted attention for offering the advantage of low adverse effects. Among these, the black mulberry plant (Morus nigra L.) stands out as a natural source of polyphenols, widely used to treat metabolic dysfunctions and confer benefits on human health. This study explores the potential of this plant as an anticancer agent, examining its effectiveness based on the type of application of the plant extracts or isolated substances, extraction methods, and its potential biological effects on cancer cells. Consequently, this study contributes to a better understanding of the distribution of phytochemicals in M. nigra and their applications in the context of cancer field. Among the compounds found in black mulberry are flavonoids, chlorogenic acid, cryptochlorogenic acid, and protocatechuic acid, along with cyanidin-3-O-glucoside as the main anthocyanin on the fruit. The phytochemicals derived from M. nigra exhibit antinociceptive and antimicrobial activities, while also showing protective effects, such as antioxidant properties that underline their potential as anticancer agents. The black mulberry's roots, stem bark, pulp, and leaves are particularly rich sources of anti-inflammatory compounds. Ethanol and methanol extraction methods appear to be the most effective in cancer management, offering compounds that facilitate the integration of apoptosis induction, cell growth inhibition, and cytotoxicity modulation. These results collectively represent the salient biological attributes that positioned black mulberry as a promising anticancer agent. Therefore, these findings highlight the multifaceted potential of M. nigra as an anticancer agent, making a compelling case for further research to advance prospects in the medical field.

Oxidative stress is a primary contributor to cerebral ischemia/reperfusion (CI/R) injury, and the use of antioxidants represents a crucial therapeutic strategy for managing CI/R injury. This study aims to explore the antioxidant effects of benzyl ferulate on CI/R injury and elucidate its underlying mechanisms. In vivo models of CI/R injury and hypoxia/reoxygenation (H/R) injury in SH-SY5Y cells were established, followed by treatment with benzyl ferulate. The extent of oxidative stress was assessed through evaluations of neurobiological function, cerebral infarct volume, reactive oxygen species (ROS), apoptosis levels, etc. Results indicated that benzyl ferulate significantly downregulated the expression of NADPH oxidase 2 (NOX) 2/NOX4 while upregulating miRNAs (652/532/92b) in CI/R rats or SH-SY5Y cells. It also reduced total NOX enzyme activity, enhanced superoxide dismutase (SOD) activity, decreased ROS and malondialdehyde (MDA) production, and inhibited cleaved caspase-3 and Bax expression-ultimately leading to reduced cell apoptosis. Benzyl ferulate effectively mitigates oxidative stress injuries of middle cerebral artery occlusion (MCAO) rats or SH-SY5Y cells subjected to H/R, and its mechanism appears to involve modulation of the miRNAs (652/532/92b)/NOX2/4 axis. This study first proved that benzyl ferulate is a promising antioxidant candidate for treating CI/R injury.