Chemical Composition of Bawei Longzuan Granule and Its Anti‐Arthritic Activity on Collagen‐Induced Arthritis in Rats by Inhibiting Inflammatory Responses

Bawei Longzuan granule (BLG) is a representative Zhuang medicine preparation. The present work aims to characterize the chemical constituents of BLG and evaluate its anti‐arthritic activity. The major chemical constituents of BLG were tentatively identified by ultra‐performance liquid chromatography‐quadrupole time‐of‐flight mass spectrometry (UPLC‐Q‐TOF/MS), which revealed the presence of some alkaloids (e. g., magnoflorine, sinomenine and nitidine) and flavonoids (e. g., hesperidin, diosmin and sinensetin) that may be partly responsible for the anti‐arthritic effect of BLG. In addition, the collagen‐induced arthritis (CIA) model in rats was induced by intradermal injection of bovine collagen‐II in complete Freund's adjuvant at the base of tail. The CIA rats received oral administration of BLG (1.25, 2.5 and 5 g/kg) for 30 days. Then, various indicators were determined to evaluate its anti‐arthritic activity, including paw swelling, arthritic score, body weight, knee joint pathology, thymus index and spleen index. Additionally, the serum levels of tumor necrosis factor (TNF)‐α, interferon (IFN)‐γ, interleukin (IL)‐1β, IL‐6, IL‐4 and IL‐10 were measured to determine the underlying mechanisms. The results showed that BLG efficiently ameliorated the severity of arthritis in CIA rats by decreasing paw swelling and arthritis score and improving the histological lesions of knee joint. Moreover, the serum levels of several pro‐inflammatory cytokines (i. e., IL‐1β, TNF‐α, IL‐6 and IFN‐γ) were downregulated, whereas two anti‐inflammatory factors (i. e., IL‐4 and IL‐10) were upregulated after BLG administration. These results indicated that BLG possessed promising therapeutic effect on collagen‐induced arthritis by inhibiting inflammatory responses. BLG can be used as a complementary or alternative traditional medicine to treat rheumatoid arthritis.     

Investigation on Toxicity and Teratogenicity in Rats of a Retinoid‐Polyamine Conjugate with Potent Anti‐Inflammatory Properties

Previous studies have shown that N¹,N¹²‐bis(all‐trans‐retinoyl)spermine (RASP), a retinoid analog, inhibits RNase P activity and angiogenesis in the chicken embryo chorioallantoic membrane, demonstrates anti‐tumor activity on prostate cancer cells, and acts as anti‐inflammatory agent, being more effective and less toxic than all‐trans retinoic acid. In an attempt to further characterize the biological profile of RASP, we tested its effects on organ toxicity and teratogenicity by daily oral gavage of RASP at a level of 50 mg/Kg of body weight in two generations of rats. We found that this compound does not induce changes to the body growth, the appearance of physical features, and the animal's reflexes. Additionally, no substantial histopathological lesions were found in brain, heart, lung, thymus, liver, thyroid gland, adrenal gland, pituitary gland, kidneys, spleen, skin, femora, prostate, testis, epididymis, vagina, uterus, and ovaries of RASP‐treated animals. These results suggest RASP, as a promising lead compound for the treatment of several dermatological disorders and certain cancer types, has apparently minimal toxic side‐effects as revealed in this two‐generation reproduction study in rats.     

SiRNA‐Mediated Down‐Regulation of CLIC4 Gene Inhibits Cell Proliferation and Accelerates Cell Apoptosis of Mouse Liver Cancer Hca‐F and Hca‐P Cells

This study explored the effects involved in silencing CLIC4 on apoptosis and proliferation of mouse liver cancer Hca‐F and Hca‐P cells. A CLIC4‐target small interfering RNA (siRNA) was designed to compound into two individual complementary oligonucleotide chains. A process of annealing and connection to a pSilencer vector was followed by transfection with Hca‐F and Hca‐P cells. Quantitative real‐time polymerase chain reaction and Western blotting techniques were used to determine CLIC4 mRNA and protein expressions. CCK8 assay and flow cytometry were employed for analysis of the survival and apoptosis rate as well as the cell cycle in an octreotide‐induced apoptosis model. Expressions of caspase 3, caspase 9, and cleaved PARP were measured using Western blotting. The CLIC4 mRNA and protein expressions in Hca‐F and Hca‐P cells transfected by pSilencer‐CLIC4 siRNA plasmid in the blank group displayed remarkably decreased levels of expression, when compared with both the control and negative control (NC) groups. Decreased survival rates and cleaved PARP expression, increased cell apoptosis rate,expressions of caspase 3 and caspase 9 in Hca‐F and Hca‐P cells were detected in groups that had been cultured in a medium containing octreotide. The pSilencer‐CLIC4 siRNA‐2 group when compared with the control and NC groups exhibited decreased survival rates, cleaved PARP expression, increased cell apoptosis rates, and increased expressions of caspase 3 and caspase 9 of Hca‐F and Hca‐P cells. The results demonstrated that siRNA‐induced down‐regulation of CLIC4 could proliferation, while in turn promoting apoptosis of mouse liver cancer Hca‐F and Hca‐P cells. J. Cell. Biochem. 119: 659–668, 2018. © 2017 Wiley Periodicals, Inc.     

Effects of Low‐Intensity Microwave Radiation on Oxidant‐Antioxidant Parameters and DNA Damage in the Liver of Rats

The continuously increasing usage of cell phones has raised concerns about the adverse effects of microwave radiation (MWR) emitted by cell phones on health. Several in vitro and in vivo studies have claimed that MWR may cause various kinds of damage in tissues. The aim of this study is to examine the possible effects of exposure to low‐intensity MWR on DNA and oxidative damage in the livers of rats. Eighteen Sprague–Dawley male rats were divided into three equal groups randomly (n = 6). Group 1 (Sham‐control): rats were kept under conditions the same as those of other groups, except for MWR exposure. Group 2: rats exposed to 1800 MHz (SAR: 0.62 W/kg) at 0.127 ± 0.04 mW/cm² power density, and Group 3: rats exposed to 2,100 MHz (SAR: 0.2 W/kg) at 0.038 ± 0.03 mW/cm² power density. Microwave application groups were exposed to MWR 2 h/day for 7 months. At the end of the exposure period, the rats were sacrificed and DNA damage, malondialdehyde (MDA), 8‐hydroxydeoxyguanosine (8‐OHdG), and total oxidant‐antioxidant parameter analyses were conducted in their liver tissue samples. It was found that 1800 and 2100 MHz low‐intensity MWR caused a significant increase in MDA, 8‐OHdG, total oxidant status, oxidative stress index, and comet assay tail intensity (P < 0.05), while total antioxidant status levels (P < 0.05) decreased. The results of our study showed that whole‐body exposure to 1800 and 2100 MHz low‐intensity MWR emitted by cell phones can induce oxidative stress by altering oxidant‐antioxidant parameters and lead to DNA strand breaks and oxidative DNA damage in the liver of rats. Bioelectromagnetics. 2021;42:76–85. © 2020 Bioelectromagnetics Society     

Phenolic Composition,Anti‐Inflammatory,Antioxidant, and Antimicrobial Activities of Alchemilla mollis (Buser) Rothm.

The current study was designed to evaluate the antioxidant, anti‐inflammatory and antimicrobial activities of Alchemilla mollis (Buser ) Rothm . (Rosaceae) aerial parts extracts. Chemical composition was analyzed by spectrophotometric and chromatographic (HPLC) techniques. The antioxidant properties assessed included DPPH^· and ABTS^·+ radical scavenging, β‐carotene‐linoleic acid co‐oxidation assay. Antimicrobial activity was evaluated with disc diffusion and micro dilution method. In order to evaluate toxicity of the extracts, with the sulforhodamine B colorimetric assay L929 cell line (mouse fibroblast) was used. The anti‐inflammatory activities of the potent antioxidant extracts (methanol, 70% methanol, and water extracts) were determined by measuring the inhibitory effects on NO production and pro‐inflammatory cytokine TNF‐α levels in lipopolysaccharide stimulated RAW 264.7 cells. 70% methanol and water extracts which were found to be rich in phenolic compounds (184.79 and 172.60 mg GAE/g extract) showed higher antioxidant activity. Luteolin‐7‐O‐glucoside was the main compound in the extracts. Ethyl acetate and 70% methanol extracts showed higher antibacterial activity against Staphylococcus aureus and Salmonella enteritidis with MIC value of 125 μg/ml. 70% methanol extract potentially inhibited the NO and TNF‐α production (18.43 μm and 1556.22 pg/ml, respectively, 6 h).     

Interaction of Helicobacter pylori Infection and Nonsteroidal Anti‐Inflammatory Drugs in Gastric and Duodenal Ulcers

Background: Gastric (GU) and duodenal ulcers (DU) are in most instances either induced by Helicobacter pylori infection or by nonsteroidal anti‐inflammatory drugs (NSAIDs). Whether eradication of H. pylori is beneficial in NSAID users for preventing NSAID induced GU and DU has been the focus of different studies. Materials and Methods: Mechanisms shared by both H. pylori and NSAIDs for the induction of GU and DU were reviewed and randomized controlled trials on H. pylori eradication for prevention and healing of GU and DU in patients requiring NSAID therapy were identified by a PubMed search. Results: Key factors in the induction of GU and DU for both H. pylori and NSAIDs are a decrease in pH, imbalance between apoptosis and proliferation, reduction in mucosal blood flow, and recruitment of polymorphonucleates in distinct compartments. For primary ulcer prevention, H. pylori eradication before starting an NSAID therapy reduces the risk of NSAID induced GU and virtually abolishes the risk of DU. H. pylori eradication alone is not sufficient for secondary prevention of NSAID induced GU and DU. H. pylori infection appears to further increase the protective effects of proton‐pump inhibitors (PPI) to reduce the risk of ulcer relapse. H. pylori eradication does not influence the healing of both GU and DU if NSAID intake is discontinued. Conclusions: Duodenal ulcer is more closely related to H. pylori infection than GU in NSAID users. H. pylori eradication is recommended for primary prevention of GU and DU in patients requiring NSAID therapy. PPI therapy is mandatory for secondary prevention of gastroduodenal ulcers, and appears to further reduce the risk of ulcer relapse in the presence of H. pylori.     

Expressions of Leptin and Insulin‐Like Growth Factor‐I Are Highly Correlated and Region‐Specific in Adipose Tissue of Growing Rats

Objective: Anatomically distinct adipose tissue regions differ in their predominant modality of growth (i.e., cellular hypertrophy vs. hyperplasia). We examined site‐specific patterns of expression of two genes whose products, leptin and insulin‐like growth factor‐I (IGF‐I), could be involved in mediating differential growth and metabolism of white adipose tissue. We also related these patterns of expression to measures of adipose depot cellularity. Research Methods and Procedures: Male Wistar rats were fed ad libitum and studied from ages 7 weeks to ～12 months. Terminal measures of body weights; weights, composition, and cellularity of four white adipose depots; circulating leptin and IGF‐I; and adipose depot‐specific expression levels of leptin and IGF‐I were measured in subsets of rats at 7, 12, 22, 42, and 46 weeks of age. Results: Both leptin and IGF‐I mRNAs are quantitatively expressed in a depot‐specific manner, in the following order: retroperitoneal ? epididymal > mesenteric > subcutaneous inguinal. Furthermore, there is a marked correlation between the expressions of these hormones in the various regions of adipose tissue of rats during the first year of life. The mechanisms that underlie the parallel expressions of leptin and IGF‐I appear to be related to fat‐cell volume. Discussion: Because both leptin and IGF‐I have been implicated in the regulation of energy homeostasis and are both expressed in adipose tissue, the depot‐specific linkage between the two genes suggests interaction at the autocrine level. This interaction may have an important role in determining functional properties particular to individual adipose depots.     

Gender‐Related In Vitro Metabolism of Hexaconazole and Its Enantiomers in Rats

In the present study we investigated the enantioselective disappearance of hexaconazole in rat liver microsomes system prepared from both genders. High‐performance liquid chromatography (HPLC) was used for identification and quantification. The degradation of the (+)‐hexaconazole was faster than that of the (?)‐hexaconazole in racemic hexaconazole and single enantiomer incubation in both sexes. The degradation half‐life of the (+)‐hexaconazole or (?)‐hexaconazole was also gender‐related. The metabolism of (+)‐hexaconazole and (?)‐hexaconazole were faster in male rat hepatic microsomes than that in female, suggesting that at least one of the cytochrome P450s (CYP) in the male rat liver microsomes system responsible for hexaconazole metabolism was male‐specific or considerably more active. Kinetic assays showed that the intrinsic clearance in male rat liver microsomes was higher than that in female. All these results strongly suggest that sexual dimorphic metabolism of hexaconazole exists in rats. The inhibition experiments with CYP inhibitors showed that the inhibitory effect of inhibitors was enantioselective and affected by sex. The results suggest that the enantioselective metabolism of hexaconazole was determined by the amount of hepatic cytochrome P450 and the expression of individual isoforms of CYPs. Chirality 25:852–857, 2013. © 2013 Wiley Periodicals, Inc.     

Medium‐Chain Oil Reduces Fat Mass and Down‐regulates Expression of Adipogenic Genes in Rats

Objective: To test the hypothesis that adipose tissue could be one of the primary targets through which medium‐chain fatty acids (MCFAs) exert their metabolic influence. Research Methods and Procedures: Sprague‐Dawley rats were fed a control high‐fat diet compared with an isocaloric diet rich in medium‐chain triglycerides (MCTs). We determined the effects of MCTs on body fat mass, plasma leptin and lipid levels, acyl chain composition of adipose triglycerides and phospholipids, adipose tissue lipoprotein lipase activity, and the expression of key adipogenic genes. Tissue triglyceride content was measured in heart and gastrocnemius muscle, and whole body insulin sensitivity and glucose tolerance were also measured. The effects of MCFAs on lipoprotein lipase activity and adipogenic gene expression were also assessed in vitro using cultured adipose tissue explants or 3T3‐L1 adipocytes. Results: MCT‐fed animals had smaller fat pads, and they contained a considerable amount of MCFAs in both triglycerides and phospholipids. A number of key adipogenic genes were down‐regulated, including peroxisome proliferator activated receptor γ and CCAAT/enhancer binding protein α and their downstream metabolic target genes. We also found reduced adipose tissue lipoprotein lipase activity and improved insulin sensitivity and glucose tolerance in MCT‐fed animals. Analogous effects of MCFAs on adipogenic genes were found in cultured rat adipose tissue explants and 3T3‐L1 adipocytes. Discussion: These results suggest that direct inhibitory effects of MCFAs on adiposity may play an important role in the regulation of body fat development.     

Responses of the Antioxidant System in QGY‐7701 Cells to the Cytotoxicity and Apoptosis Induced by 1‐Octyl‐3‐methylimidazolium Chloride

This study aims to evaluate the cytotoxicity and responses of the cellular antioxidant system of 1‐octyl‐3‐methylimidazolium chloride ([C₈mim][Cl]) on human hepatocarcinoma QGY‐7701 cells. The results show that [C₈mim][Cl] can inhibit QGY‐7701 cell growth and decrease their viabilities in a dose‐dependent manner. The results also reveal that [C₈mim][Cl] exposure can induce apoptosis in the [C₈mim][Cl]‐treated QGY‐7701 cells. In addition, the results of biochemical assays show that [C₈mim][Cl] exposure causes overproduction of reactive oxygen species (ROS), inhibits superoxide dismutase (SOD) and catalase (CAT) activities, decreases reduced glutathione (GSH) content, and increases the cellular malondialdehyde (MDA) level. These results suggest that ROS‐mediated oxidative stress may be responsible for the apoptosis induced by [C₈mim][Cl] in QGY‐7701 cells. © 2013 Wiley Periodicals, Inc. J BiochemMol Toxicol 27:330‐336, 2013; View this article online at wileyonlinelibrary.com . DOI 10.1002/jbt.21495     

Essential Oil of Azorella cryptantha Collected in Two Different Locations from San Juan Province,Argentina: Chemical Variability and Anti‐Insect and Antimicrobial Activities

The essential oils (EOs) of two populations of Azorella cryptantha (Clos) Reiche , a native species from San Juan Province, were obtained by hydrodistillation in a Clevenger‐type apparatus and characterized by GC‐FID and GC/MS analyses. The compounds identified amounted to 92.3 and 88.7% of the total oil composition for A. cryptantha from Bauchaceta (Ac‐BAU) and Agua Negra (Ac‐AN), respectively. The EO composition for the two populations was similar, although with differences in the identity and content of the main compounds and also in the identity of minor components. The main compounds of the Ac‐BAU EO were α‐pinene, α‐thujene, sabinene, δ‐cadinene, δ‐cadinol, trans‐β‐guaiene, and τ‐muurolol, while α‐pinene, α‐thujene, β‐pinene, γ‐cadinene, τ‐cadinol, δ‐cadinene, τ‐muurolol, and a not identified compound were the main constituents of the Ac‐AN EO, which also contained 3.0% of oxygenated monoterpenes. The repellent activity on Triatoma infestans nymphs was 100 and 92% for the Ac‐AN and Ac‐BAU EOs, respectively. Regarding the toxic effects on Ceratitis capitata, the EOs were very active with LD₅₀ values lower than 11 μg/fly. The dermatophytes Microsporum gypseum, Trichophyton rubrum, and T. mentagrophytes and the bacterial strains Escherichia coli LM₁, E. coli LM₂, and Yersinia enterocolitica PI were more sensitive toward the Ac‐AN EO (MIC 125 μg/ml) than toward the Ac‐BAU EO. This is the first report on the composition of A. cryptantha EO and its anti‐insect and antimicrobial properties.