Pyrrolizidine alkaloids (PAs) are a group of naturally occurring alkaloids widely present in plants. PAs are highly hepatotoxic and have been documented to cause many incidents of human and animal ...poisoning. Retrorsine (RTS) is a pyrrolizidine alkaloid (PA) derived from the Compositae Senecio, which has been shown to cause hepatotoxicity. Human liver poisoning occurs through the consumption of RTS-contaminated food, and animals can also be poisoned by ingesting RTS-containing toxic plants. The mechanism of RTS-induced liver toxicity is not fully understood. In this study, we demonstrated that RTS-induced oxidative stress plays a pivotal role in RTS-induced liver toxicity involving apoptosis and autophagy. The results showed that RTS treatment in the cultured Primary rat hepatocytes caused cytotoxicity and release of aspartate aminotransferase (AST) and alanine aminotransferase (ALT) in a time- and dose-dependent manner. Our study showed that treatment of RTS induced ROS and MDA (malondialdehyde, a lipid peroxidation marker) in the hepatocytes, and reduced antioxidant capacity (GSH content, SOD activity), suggesting RTS treatment caused oxidative stress response in the hepatocytes. Furthermore, we found that RTS induced apoptosis and autophagy in the hepatocytes, and RTS-induced apoptosis and autophagy could be alleviated by ROS scavenger N-acetylcysteine (NAC) and the MAPK pathway inhibitors suggesting ROS/MAPK signaling pathway plays a role in RTS induced apoptosis and autophagy. Collectively, this study reveals the regulatory mechanism of oxidative stress in RTS-induced apoptosis and autophagy in the hepatocytes, providing important insights of molecular mechanisms of hepatotoxicity induced by RTS and related pyrrolizidine alkaloids in liver. This mechanism provides a basis for the prevention and treatment of PA poisoning in humans and animals.
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•Retrorsine could induce the release of AST and ALT, and exhibit significant hepatotoxicity.•Retrorsine could induce oxidative stress and apoptosis and autophagy in hepatocytes.•Retrorsine could regulate the apoptosis and autophagy of hepatocytes through the ROS/MAPK signaling pathway.
This animal study aimed to explore the effects of patellar hypermobility and patellar dislocation on the developing femoral trochlea.
Seventy-two 3-week-old Wistar rats were randomly divided into ...three groups. The sham group (SG) underwent simple incision and suture of the skin and subcutaneous tissue; the patellar hypermobility group (PHG) underwent medial and lateral retinacular release and pie-crusting technique for the patellar ligament; the patellar dislocation group (PDG) underwent plication of the medial patellofemoral retinaculum. Twelve rats in each group were euthanized at 3 and 6 weeks postoperatively, respectively, and specimens were collected. The bony sulcus angle (BSA), cartilaginous sulcus angle (CSA), trochlear sulcus depth (TSD), and thickness of the cartilage on the lateral facet (CTL), medial facet (CTM), and center (CTC) of the trochlea were measured on hematoxylin and eosin-stained sections.
In the PHG and PDG, the femoral condyles became blunt, the trochlear groove became shallower, and cartilage became thicker compared with the SG. Compared with the SG, the PHG and PDG had significantly larger BSA and CSA values at 3 (p < 0.05) and 6 weeks (p < 0.005), and a significantly shallower TSD (p < 0.05). At 3 weeks, all cartilage thicknesses in the PHG and the CTC and CTM in the PDG were significantly thinner than in the SG (PHG vs. SG: p = 0.009 for CTL, p < 0.001 for CTM, p = 0.003 for CTC; PDG vs. SG: p = 0.028 for CTC, p = 0.048 for CTM). At 6 weeks, the CTC was thicker in the PHG and PDG than the SG (PHG vs. SG: p = 0.044; PDG vs. SG: p = 0.027), and the CTL was thinner in the PDG than the SG (p = 0.044).
Patellar hypermobility and patellar dislocation may result in trochlear dysplasia that worsens with age. Excessive or insufficient loading leads to trochlear dysplasia.
The relationship between breech presentation and trochlear dysplasia has been confirmed. However, the pathological process of breech-related trochlear dysplasia remains unclear. This study aimed to ...establish an animal model to simulate breech presentation and to analyze the pathological process of the femoral trochlea.
One hundred and twenty neonatal rats were randomly assigned into a control group and two experimental groups that were swaddled (using surgical tape) to keep the hip flexed and knees extended to simulate human breech presentation for the 5 days (short Swaddling) and the 10 days (prolonged Swaddling) of life. Gross and cross-sectional observation, histological staining measurement in two experimental time points (5 and 10 days after birth) were conducted to evaluate the morphological changes of the femoral trochlea.
The incidence of trochlear dysplasia increased with the Swaddling time. Rats in the prolonged Swaddling group had the high prevalence of trochlea dysplasia (52 of 60), followed by short Swaddling group (42 of 60). Gross and cross-sectional observation showed a shallower trochlea groove in two experimental groups. Histologicalstaining measurement indicated that the trochlear sulcus angle and trochlear sulcus depth were significantly different between the experimental group and the control group since day 5 and day 10.
In this model, breech presentation had an adverse effect on neonatal knees and could induce trochlear dysplasia. In addition, this study also showed that the more time in breech presentation, the more incidence of trochlear dysplasia.
Artemisia is important medicinal plants in China and are widely used in medicine, agriculture, and food. Pharmacologically active components of the plants remain to be investigated.
This study sought ...to identify and compare the chemical constituents of three species of Artemisia in Tibet using a widely-targeted metabolomics approach and their antibacterial and antioxidant capacities were determined.
A total of 1109 metabolites within 10 categories were detected from the three species of Artemisia, including lipids, amino acids, nucleotides, flavonoids, terpenes, coumarins, organic acids, and phenolic acids. 732 different metabolites have been identified between Artemisia sieversiana and Artemisia annua, 751 different metabolites were identified between Artemisia wellbyi and A. sieversiana, and 768 differential metabolites were differentially detected from A. wellbyi and A. annua. Differentially identified compounds included flavonoids, phenolic acids, artemisinins and coumarin. A. annua contained the highest relative content of artemisinin among three Artemisia. The antimicrobial experiments showed that the three Artemisia species had strong antibiotic activities against Bacillus subtilis, Escherichia coli, Staphylococcus aureus, Proteus mirabilis and Pseudomonas aeruginosa. The biochemical analysis showed that the three species of Artemisia have strong antioxidant capacity.
This is the first reported attempt to comparatively determine the types of the metabolites of the three widely distributed Artemisia species in Tibet. The information should help medicinal research and facilitate comprehensive development and utilization of Artemisia species in Tibet.
We aimed to investigate the role and working mechanism of Homo sapiens circular RNA_0003602 (hsa_circ_0003602) in colorectal cancer (CRC) development.
The expression of circ_0003602, miR-149-5p, and ...solute carrier family 38 member 1 (SLC38A1) was detected by quantitative real-time polymerase chain reaction. RNase R assays were conducted to determine the characteristics of circ_0003602. CCK-8 assays, flow cytometry analysis, transwell invasion assays, wound healing assays and tube formation assays were employed to evaluate cell viability, apoptosis, invasion, migration, and angiogenesis. All protein levels were examined by Western blot or immunohistochemistry assay. The glutamine metabolism was monitored by corresponding glutamine, α-ketoglutarate and glutamate assay kits. Dual-luciferase reporter assay was utilized to confirm the targeted combination between miR-149-5p and circ_0003602 or SLC38A1. A xenograft tumor model was established to analyze the role of circ_0003602 in CRC tumor growth in vivo.
Circ_0003602 was upregulated in CRC tissues and cell lines. Circ_0003602 silencing suppressed CRC cell viability, migration, invasion, angiogenesis, and glutaminolysis; induced cell apoptosis in vitro; and blocked tumor growth in vivo. Moreover, circ_0003602 directly interacted with miR-149-5p to negatively regulate its expression, and circ_0003602 knockdown suppressed the malignant behaviors of CRC cells largely by upregulating miR-149-5p. MiR-149-5p directly bound to the 3' untranslated region of SLC38A1 to induce its degradation, and miR-149-5p overexpression reduced the malignant potential of CRC cells largely by downregulating SLC38A1. Circ_0003602 positively regulated SLC38A1 expression by sponging miR-149-5p in CRC cells.
Circ_0003602 knockdown impedes CRC development by targeting the miR-149-5p/SLC38A1 axis, which provides a novel theoretical basis and new insights for CRC treatment.
Slow transit constipation (STC), the most common type of constipation, seriously affects the life of patients. Zhizhu decoction (ZZD), a traditional Chinese medicine compound, has is effective ...against functional constipation, but the mechanism is still unclear.
This research explores the mechanism of ZZD on STC from the perspective of metabolomics and gut microbiota.
Fifty-four C57BL/6 mice were randomly divided into six groups (n = 9): control (control); STC (model); positive control (positive); low-dose (5 g/kg; L-ZZD), medium-dose (10 g/kg; M-ZZD), and high-dose (20 g/kg; H-ZZD) ZZD treatment. Following treatment of mice with ZZD for two weeks, the changes in intestinal motility, colon histology, intestinal neurotransmitters, and aryl hydrocarbon receptor (AHR) pathway determined the effects of ZZD on the pathophysiology of STC. LC-MS targeting serum metabolomics was used to analyze the regulation of ZZD on neurotransmitters, and 16S rRNA high-throughput sequencing was used to detect the regulation of the gut microbiome.
ZZD had the highest content of naringin (6348.1 mg/L), and could significantly increase the 24 h defecations (1.10- to 1.42-fold), fecal moisture (1.14-fold) and intestinal transport rate (1.28-fold) of STC mice, increased the thickness of the mucosal and muscular tissue (1.18- to 2.16-fold) and regulated the neurotransmitters in the colon of STC mice. Moreover, ZZD significantly activated the AHR signaling pathway, and also affected the composition of gut microbiota in STC mice.
The beneficial effect and the possible mechanism of ZZD on STC could provide a theoretical basis for the broader clinical application of ZZD.
Colorectal cancer is one of the most common cancers, and its incidence rate keeps increasing worldwide. Epidermal growth factor receptor (EGFR) is highly expressed in colorectal cancer cells, and its ...activation leads to the activation of downstream kinase pathways to promote cell growth, resulting in tumour growth and progression. EGFR is expressed in only small amounts in normal cells, however, it is overexpressed in tumour cells and is a potential tumour marker, thus the detection of which is of great importance. This work proposes a novel and highly sensitive electrochemical immunosensor based on nucleic acid aptamer recognition and silver deposition for EGFR detection. The key novelty of this work is the use of SiO2 nanospheres to enhance the sensitivity. First, SiO2 nanospheres were synthesized and modified on the surface of a gold electrode using a self-assembly process. This provided a large surface area to allow high loading of capture antibodies. The antibody was then immobilized on the SiO2 nanosphere modified electrode surface to form an immunosandwich complex with the antigen and biotin-labelled nucleic acid aptamer. Afterwards, the affinity-labelled alkaline phosphatase was immobilized onto the electrode with the specific reaction of biotin and affinity. The obtained product was subjected to silver deposition in the substrate solution. The large surface area provided by the SiO2 nanospheres allowed significantly improved sensitivity. Experiments were performed to optimize the concentration of immobilized antibodies and biotin-labelled nucleic acid aptamer. The experimental results show a good linearity of EGFR concentration in the range of 1 to 1000 ng/mL, with the lowest detection limit up to 0.06 ng/mL. This method has high sensitivity and stability, which allows it to have a broad application prospects in the clinical field.
Background/Aims: We aimed to investigate the role and working mechanism of Homo sapiens circular RNA_0003602 (hsa_circ_0003602) in colorectal cancer (CRC) development.
Methods: The expression of ...circ_0003602, miR-149-5p, and solute carrier family 38 member 1 (SLC38A1) was detected by quantitative real-time polymerase chain reaction. RNase R assays were conducted to determine the characteristics of circ_0003602. CCK-8 assays, flow cytometry analysis, transwell invasion assays, wound healing assays and tube formation assays were employed to evaluate cell viability, apoptosis, invasion, migration, and angiogenesis. All protein levels were examined by Western blot or immunohistochemistry assay. The glutamine metabolism was monitored by corresponding glutamine, α-ketoglutarate and glutamate assay kits. Dualluciferase reporter assay was utilized to confirm the targeted combination between miR-149-5p and circ_0003602 or SLC38A1. A xenograft tumor model was established to analyze the role of circ_0003602 in CRC tumor growth in vivo.
Results: Circ_0003602 was upregulated in CRC tissues and cell lines. Circ_0003602 silencing suppressed CRC cell viability, migration, invasion, angiogenesis, and glutaminolysis; induced cell apoptosis in vitro; and blocked tumor growth in vivo. Moreover, circ_0003602 directly interacted with miR-149-5p to negatively regulate its expression, and circ_0003602 knockdown suppressed the malignant behaviors of CRC cells largely by upregulating miR-149-5p. MiR-149-5p directly bound to the 3’ untranslated region of SLC38A1 to induce its degradation, and miR-149-5p overexpression reduced the malignant potential of CRC cells largely by downregulating SLC38A1. Circ_0003602 positively regulated SLC38A1 expression by sponging miR-149-5p in CRC cells.
Conclusions: Circ_0003602 knockdown impedes CRC development by targeting the miR-149- 5p/SLC38A1 axis, which provides a novel theoretical basis and new insights for CRC treatment. (Gut Liver 2023;17:267-279)
Patellar dislocation can cause a series of changes in the trochlear groove and patella. However, the influence of patellar dislocation on the medialis (VM) and vastus lateralis (VL) muscles and ...whether nerve growth factor (NGF) is beneficial to proprioceptive rehabilitation for patellar dislocation are unknown. The purpose of this study was to investigate the effects on VM and VL after the injection of NGF and early reduction in rabbits for patellar dislocation with electrophysiological and pathological analysis.
Sixty 2-month-old rabbits were randomly divided into four groups (15 rabbits in each group). Rabbits in Group 1, Group 2, and Group 3 underwent patellar dislocation surgery, and rabbits in Group 4 underwent sham surgery. One month later, patellar reduction was performed in Groups 1 and 2. NGF was injected into the rabbits of Group 1. The electrophysiological and pathological changes in VM and VL were analyzed at 1 month and 3 months after patellar reduction.
The electrophysiological and pathological indices in Groups 1 and 2 were significantly different from those in Group 3 at 1 and 3 months after patellar reduction. There were significant differences between NGF injection Group 1 and Group 2 without NGF injection. There was no significant difference between Group 1 and Group 4 at 3 months after patellar reduction.
Patellar dislocation can cause abnormal electrophysiological and pathological effects on VM and VL. Patellar reduction should be performed as early as possible, and NGF injection may be beneficial to the rehabilitation of proprioception.
Fetal alcohol spectrum disorder (FASD) is the leading known cause of intellectual disability, and may manifest as deficits in cognitive function, including working memory. Working memory capacity and ...accuracy increases during adolescence when neurons in the prefrontal cortex undergo refinement. Rats exposed to low doses of ethanol prenatally show deficits in working memory during adolescence, and in cognitive flexibility in young adulthood. The cholinergic system plays a crucial role in learning and memory processes. Here we report that the combination of choline and training on a working memory task during adolescence significantly improved cognitive flexibility (performance on an attentional set shifting task) in young adulthood: 92% of all females and 81% of control males formed an attentional set, but only 36% of ethanol-exposed males did. Resting state functional magnetic resonance imaging showed that functional connectivity among brain regions was different between the sexes, and was altered by prenatal ethanol exposure and by choline + training. Connectivity, particularly between prefrontal cortex and striatum, was also different in males that formed a set compared with those that did not. Together, these findings indicate that prenatal exposure to low doses of ethanol has persistent effects on brain functional connectivity and behavior, that these effects are sex-dependent, and that an adolescent intervention could mitigate some of the effects of prenatal ethanol exposure.
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