Beneficial effect of honokiol and magnolol on polyol pathway and oxidative stress parameters in the testes of diabetic rats.

In diabetes hyperglycemia, excessive production of free radicals and present oxidative stress lead to many complications in the body, including male reproductive system disorders. To prevent the development of diabetic complications in the testes resulting from them, it seems beneficial to include compounds considered as natural antioxidants. Honokiol and magnolol are neolignans obtained from magnolia bark, which possess proven antioxidant properties. The aim of this study was to evaluate the effect of honokiol and magnolol on the parameters of oxidative stress, polyol pathway and glycation products in the testes as well as on selected biochemical parameters in the blood serum of rats with type 2 diabetes. The study was conducted on mature male Wistar rats with high fat diet and streptozotocin-induced type 2 diabetes. Neolignans-treated rats received honokiol or magnolol orally at the doses of 5 or 25 mg/kg, respectively, for 4 weeks. Parameters related to glucose and lipid homeostasis, basic serological parameters and sex hormones level in the serum as well as polyol pathway parameters, antioxidant enzyme activity, endogenous antioxidants level, sumaric parameters for oxidative stress and oxidative damage in the testes were estimated. Oral administration of honokiol and magnolol turned out to be beneficial in combating the effects of oxidative stess in the testes, but showed no favorable effects on serum biochemical parameters. Additionally, magnolol compared to honokiol revealed more advantageous impact indicating the reversal of the effects of diabetic complications in the male reproductive system and counteracted oxidative stress damages and polyol pathway disorders in the testes.

Effects of Water-Ethanol Extracts from Four Sphagnum Species on Gene Expression of Selected Enzymes in Normal Human Dermal Fibroblasts and Their Antioxidant Properties.

Mosses (Bryophyta), particularly species of the genus Sphagnum, which have been used for centuries for the treatment of skin diseases and damage, are still not explored enough in terms of their use in cosmetics. The purpose of this study was to determine the antioxidant properties of water-ethanol extracts from four selected species of the genus Sphagnum (S. girgenshonii Russow, S. magellanicum Brid., S. palustre L., and S. squarrosum Crome) and their impact on the expression of genes encoding key enzymes for the functioning of the skin. In this study, the effects of Sphagnum extracts on the expression of genes encoding tyrosinase, collagenase, elastase, hyaluronidase and hyaluronic acid synthase in human dermal fibroblasts were determined for the first time in vitro. The extracts inhibited tyrosinase gene expression and showed antioxidant activity. The experiment showed an increase in the expression of some genes encoding collagenase (MMP1) or hyaluronidase (HYAL2, HYAL3 and HYAL4) and a decrease in the hyaluronan synthase (HAS1, HAS2 and HAS3) genes expression by the tested extracts. The obtained results suggest that using extracts from the tested Sphagnum species in anti-aging cosmetics does not seem beneficial. Further studies are needed to clarify their impact on the skin.

Mapping of DNA methylation-sensitive cellular processes in gingival and periodontal ligament fibroblasts in the context of periodontal tissue homeostasis.

Interactions between gingival fibroblasts (GFs) and oral pathogens contribute to the chronicity of inflammation in periodontitis. Epigenetic changes in DNA methylation are involved in periodontitis pathogenesis, and recent studies indicate that DNA methyltransferase (DNMT) inhibitors may protect against epithelial barrier disruption and bone resorption. To assess the impact of DNMT inhibition on GFs, cells were cultured with decitabine (5-aza-2'-deoxycytidine, DAC) for 12 days to induce DNA hypomethylation. We observed several potentially detrimental effects of DAC on GF biological functions. First, extended treatment with DAC reduced GF proliferation and induced necrotic cell death. Second, DAC amplified Porphyromonas gingivalis- and cytokine-induced expression and secretion of the chemokine CCL20 and several matrix metalloproteinases (MMPs), including MMP1, MMP9, and MMP13. Similar pro-inflammatory effects of DAC were observed in periodontal ligament fibroblasts. Third, DAC upregulated intercellular adhesion molecule-1 (ICAM-1), which was associated with increased P. gingivalis adherence to GFs and may contribute to bacterial dissemination. Finally, analysis of DAC-induced genes identified by RNA sequencing revealed increased expression of CCL20, CCL5, CCL8, CCL13, TNF, IL1A, IL18, IL33, and CSF3, and showed that the most affected processes were related to immune and inflammatory responses. In contrast, the genes downregulated by DAC were associated with extracellular matrix and collagen fibril organization. Our observations demonstrate that studies of DNMT inhibitors provide important insights into the role of DNA methylation in cells involved in periodontitis pathogenesis. However, the therapeutic potential of hypomethylating agents in periodontal disease may be limited due to their cytotoxic effects on fibroblast populations and stimulation of pro-inflammatory pathways.

Silymarin from Milk Thistle Fruits Counteracts Selected Pathological Changes in the Lenses of Type 1 Diabetic Rats.

Diabetes is a metabolic disease affecting many tissues and organs. The main etiological factor for diabetic complications is hyperglycemia and subsequent pathologies, such as oxidative stress. One of the organs susceptible to the development of diabetic complications is the eye with all of its elements, including the lens. The aim of this study was to evaluate the effect of silymarin, an extract obtained from milk thistle fruit husks, on the oxidative stress markers in the lenses of type 1 diabetic rats. The study was performed on male rats in which type 1 diabetes was induced with 60 mg/kg streptozotocin injection. Diabetic animals were treated via an intragastric tube with silymarin at 50 and 100 mg/kg doses for four weeks. Multiple oxidative stress and polyol pathway-related parameters were measured in the lenses, and auxiliary biochemical tests in the serum were conducted. Diabetes induced severe pathological changes both in the lenses and the serum, and silymarin counteracted several of them. Nevertheless, the qualitative analyses encompassing all tested parameters indicate that silymarin slightly improved the overall state of diabetic animals. Upon the obtained results, it can be concluded that silymarin reveals a faint positive effect on the lenses in type 1 diabetic rats.

The impact of the co-exposure of melanoma cells to chlorogenic acid and a moderate-strength static magnetic field.

A static magnetic field (SMF) or the bioactive compounds that are found in foods are potential agents that can be used to support cancer therapy. Therefore, the aim of our study was to assess the impact of the SMF that are induced by neodymium magnets on the culture growth and antioxidant status of melanoma cells that had been treated with chlorogenic acid (CGA). The melanoma cells, the control and those that had been treated with CGA, were put in special magnetic test chambers that generated a 0.7 T magnetic field. The mRNA levels of the antioxidant enzymes were analyzed using RT-qPCR. The activity of SOD, GPx, and CAT was measured in the cell lysates. While the expression and activity of the antioxidant enzymes was inhibited relative to the untreated cells as a result of the CGA treatment (1 mmol/L), it was not after the CGA treatment in combination with an SMF. The demonstrated cytotoxicity of CGA (1 mmol/L) and its inhibition of the antioxidant enzymes suggests the usefulness of phenolic compounds as a supporting pharmacological treatment for melanoma. PRACTICAL APPLICATIONS: Phenolic acids and their derivatives, which are the bioactive components of the human diet, are signal molecules that transfer information from the external environment that affects the level of gene expression in cells. This study suggests the usefulness of phenolic compounds as a supporting pharmacological treatment for melanoma and seems to be important for the development of experimental oncology.

Rosmarinic and Sinapic Acids May Increase the Content of Reduced Glutathione in the Lenses of Estrogen-Deficient Rats.

Oxidative stress is believed to be associated with both postmenopausal disorders and cataract development. Previously, we have demonstrated that rosmarinic and sinapic acids, which are diet-derived antioxidative phenolic acids, counteracted some disorders induced by estrogen deficiency. Other studies have shown that some phenolic acids may reduce cataract development in various animal models. However, there is no data on the effect of phenolic acids on oxidative stress markers in the lenses of estrogen-deficient rats. The study aimed to investigate whether administration of rosmarinic acid and sinapic acid affects the antioxidative abilities and oxidative damage parameters in the lenses of estrogen-deficient rats. The study was conducted on three-month-old female Wistar rats. The ovariectomized rats were orally treated with rosmarinic acid at doses of 10 and 50 mg/kg or sinapic acid at doses of 5 and 25 mg/kg, for 4 weeks. The content of reduced glutathione (GSH), oxidized glutathione and amyloid ß1-42, as well as products of protein and lipid oxidation, were assessed. Moreover, the activities of superoxide dismutase, catalase, and some glutathione-related enzymes in the lenses were determined. Rosmarinic and sinapic acids in both doses resulted in an increase in the GSH content and glutathione reductase activity. They also improved parameters connected with protein oxidation. Since GSH plays an important role in maintaining the lens transparency, the increase in GSH content in lenses after the use of rosmarinic and sinapic acids seems to be beneficial. Therefore, both the investigated dietary compounds may be helpful in preventing cataract.

Induction of Apoptosis in Normal Human Dermal Fibroblasts Infected with Borrelia burgdorferi Sensu Lato.

The spirochete Borrelia burgdorferi s.l. can enter into different eukaryotic cells. Intracellular localization of bacteria may cause many changes in different cell pathways like apoptosis-mediated caspase cascade. The present studies focused on gene expression associated with caspase cascade after normal human dermal fibroblasts (NHDF) infection with Borrelia garinii, Borrelia afzelii, and B. burgdorferi s.s. The use of oligonucleotide microarray technique enabled an expression level comparison of genes associated with caspase cascade in NHDF infected with spirochetes. The increased expression of genes associated with caspase cascade was observed in case of CASP5, CASP2, CARD10, CASP10, MALT1, and NLRP1. The decreased expression was observed in case of CASP4, CASP6, and CASP1. The mRNA expression for CASP3 was inhibited in cells infected with three genospecies of Borrelia. However, the intensity of fluorescence was not statistically significant. In addition, cell cultures were fixed and procedure of caspase-3 detection and the TUNEL assay were performed. The in situ caspase-3 detection procedure confirmed the results obtained from microarray analyses. Only several fluorescent signals were observed. Many apoptotic cells were detected in NHDF-infected cultures with all spirochete genospecies found using the TUNEL reaction.

Expression Profiles of Toll-Like Receptors in the Differentiation of an Infection with Borrelia burgdorferi Sensu Lato Spirochetes.

The similarity of Lyme borreliosis to other diseases and its complex pathogenesis present diagnostic and therapeutic difficulties. The changes that occur at the cellular and molecular levels after a Borrelia sp. infection still remain poorly understood. Therefore, the present study focused on the expression of TLR and TLR-signaling genes in human dermal fibroblasts in the differentiation of an infection with Borrelia burgdorferi sensu lato spirochetes. Normal human dermal fibroblasts were cultured with the spirochetes of Borrelia burgdorferi sensu stricto, Borrelia afzelii and Borrelia garinii. Total RNA was extracted from the cells using TRIzol reagent. The analysis of the expression profiles of TLRs and TLR-related genes was performed using commercially available oligonucleotide microarrays of HG-U133A. The GeneSpring 12.0 platform and significance analysis of microarrays were used for the statistical analysis of microarray data. The analyses using the oligonucleotide microarray and QRT-PCR techniques permitted to identify the genes encoding TLR4 and TLR6 as specific for infection with B. afzelii and B. burgdorferi sensu stricto. In turn, TLR3 was only characteristic for an infection with B. burgdorferi sensu stricto. There were no changes in the TLR gene expression after infection with B. garinii. Our findings confirm that Borrelia has a major effect on fibroblast gene expression. Further characterization of changes in gene expression may lead to valuable insights into the role of the toll-like receptor in the pathogenesis of Lyme disease and may provide guidelines for the development of diagnostic markers for an infection with a particular Borrelia genospecies. Moreover, this will help to identify better treatment strategies for Lyme disease.

BIOCHANIN A SHOWS NO EFFECT ON SKELETAL SYSTEM IN OVARIECTOMIZED RATS, WHEN ADMINISTERED IN MODERATE DOSE.

Biochanin A is a naturally occurring isoflavone. Its main sources are clover species such as Trifolium pretense, Trifolium subterraneum or Trifolium incarnatum. Phytoestrogens, including isoflavones, are plant-derived substances, which exhibit estrogen-like properties, thus they may be used as an alternative for hormonal replacement therapies and prevent postmenopausal osteoporosis. Therefore, the aim of the presented study, was to investigate the effect of biochanin A on chemistry and mechanical properties of skeletal system in rats with ovariectomy-induced osteoporosis. The animals were divided into 4 groups--(I) sham-operated rats, (II) ovariectomized rats, (III) ovariectomized rats receiving estradiol at a dose of 0.2 mg/kg p.o., which were a positive control, and (IV) ovariectomized rats receiving biochanin A at a dose of 5 mg/kg p.o. for four weeks. The administered dose of biochanin A is considered as moderate for human, which can be received in the dietary supplements, and was established using ten-fold conversion rate resulting from faster metabolism in rats. Obtained results showed that ovariectomy induced harmful changes in bone tissue, causing worsening in both chemistry and mechanical parameters in bones. Administration of biochanin A to ovariectomized rats did not affect any changes in bone tissue in comparison to the bones of untreated ovariectomized rats. There was neither improvement nor deterioration noted in chemical composition and mechanical properties in all analyzed bones. Basing on the results, it could be concluded, that biochanin A administered in a moderate dose shows no influence on bone tissue of rats with ovariectomy-induced osteoporosis.