ABSTRACT
Engineered probiotics can serve as therapeutics based on their ability of produce recombinant immune-stimulating properties. In this study, we built the recombinant Bacillus subtilis WB800 expressing antimicrobial peptide KR32 (WB800-KR32) using genetic engineering methods and investigated its protective effects of nuclear factor-E2-related factor 2 (Nrf2)-Kelch-like ECH-associated protein 1 (Keap1) pathway activation in intestinal oxidative disturbance induced by enterotoxigenic Escherichia coli (ETEC) K88 in weaned piglets. Twenty-eight weaned piglets were randomly distributed into four treatment groups with seven replicates fed with a basal diet. The feed of the control group (CON) was infused with normal sterilized saline; meanwhile, the ETEC, ETEC+WB800, and ETEC+WB800-KR32 groups were orally administered normal sterilized saline, 5×1010 CFU (CFU: colony forming units) WB800, and 5×1010 CFU WB800-KR32, respectively, on Days 1‒14 and all infused with ETEC K88 1×1010 CFU on Days 15‒17. The results showed that pretreatment with WB800-KR32 attenuated ETEC-induced intestinal disturbance, improved the mucosal activity of antioxidant enzyme (catalase (CAT), superoxide dismutase (SOD), and glutathione peroxidase (GPx)) and decreased the content of malondialdehyde (MDA). More importantly, WB800-KR32 downregulated genes involved in antioxidant defense (GPx and SOD1). Interestingly, WB800-KR32 upregulated the protein expression of Nrf2 and downregulated the protein expression of Keap1 in the ileum. WB800-KR32 markedly changed the richness estimators (Ace and Chao) of gut microbiota and increased the abundance of Eubacterium_rectale_ATCC_33656 in the feces. The results suggested that WB800-KR32 may alleviate ETEC-induced intestinal oxidative injury through the Nrf2-Keap1 pathway, providing a new perspective for WB800-KR32 as potential therapeutics to regulate intestinal oxidative disturbance in ETEC K88 infection.
Subject(s)
Animals , Swine , Enterotoxigenic Escherichia coli , Kelch-Like ECH-Associated Protein 1 , Bacillus subtilis , NF-E2-Related Factor 2 , Antioxidants , Oxidative StressABSTRACT
Objective: To investigate the antioxidative and antidiabetic effects of Harpephyllum caffrum bark infusion as well as its effects on glucogenic and nucleotide hydrolyzing enzyme activities in FeSO 4 - induced oxidative stress in rat hepatic tissue. Methods: Harpephyllum caffrum infusion was prepared from dried plant materials (40 g) infused in boiling water (400 mL) for 20 min at room temperature. The antioxidative and inhibitory activities against carbohydrate digestive enzymes of the infusion were determined using established protocols. The liver tissues of rats were used for glucose uptake assay and to evaluate the infusion's effect on endogenous antioxidant, glucogenic, and nucleotide hydrolyzing enzyme activities in FeSO 4 -induced hepatic injury. Results: The Harpephyllum caffrum infusion significantly reduced ferric iron (FRAP) and free radicals (OH • and DPPH) in a dose- dependent manner. It inhibited -amylase and -glucosidase activities and increased glucose uptake in hepatic tissues. FeSO 4 significantly decreased glutathione concentration, catalase, and superoxide dismutase activities while increasing malondialdehyde level, glycogen phosphorylase, fructose-1,6-bisphosphatase, and adenosine triphosphatase activities. However, treatment with Harpephyllum caffrum infusion reversed FeSO 4 -induced changes. Characterization of the infusion revealed the presence of catechol, O-pyrocatechuic acid, mequinol, maltol, and glycoside derivatives. Conclusions: The Harpephyllum caffrum infusion demonstrates antidiabetic and antioxidative potentials in in vitro models of type 2 diabetes as depicted by its ability to inhibit carbohydrate digestive enzymes, mitigate oxidative imbalance, and regulate glucogenic and nucleotide hydrolyzing enzyme activities in oxidative hepatic injury.
ABSTRACT
ObjectiveTo investigate the effect of calycosin-mediated glucoprotein130/Janus kinase/signal transducer and activator of transcription factor （GP130/JAK/STAT） signaling pathway on oxidative injury of astrocytes in spinal cord. MethodAstrocytes in rat spinal cord were isolated and identified by immunofluorescence detection of glial fibrillary acidic protein （GFAP）. The cells were respectively pre-treated with 5， 10， 20 μmol·L-1 calycosin for 12 h， and then 100 μmol·L-1 H2O2 （24 h） was added to induce oxidative injury. Cell counting kit-8 （CCK-8） assay was employed to detect cell proliferation and select the optimal concentration of calycosin. The following experimental groups were designed： control group， model group （100 μmol·L-1 H2O2）， calycosin group （20 μmol·L-1 calycosin）， calycosin + LY294002 group （20 μmol·L-1 calycosin + 10 μmol·L-1 LY294002）， and calycosin + Stattic group （20 μmol·L-1 calycosin + 3 μmol·L-1 Stattic）. CCK-8 assay and immunofluorescence method were used to detect the proliferation of cells and flow cytometry was applied to detect cell apoptosis and cycle. The protein expression of phosphorylated （p）-JAK2， p-STAT3， p-protein kinase B （Akt）， GP130， and interleukin-6 （IL-6） was detected by Western blotting. ResultCompared with the control group， the model group showed low proliferation activity and high apoptosis rate of cells （P<0.05）. Compared with the model group， calycosin （20 μmol·L-1） group displayed high proliferation activity and low apoptosis rate of cells （P<0.05）. Compared with calycosin （20 μmol·L-1） group， both phosphatidylinosirtol-3-kinases （PI3K） inhibitor LY294002 and STAT3 inhibitor Stattic significantly reduced the proliferation activity and increased the apoptosis rate of cells （P<0.05）. The protein expression of p-JAK2， p-STAT3， p-Akt， GP130， and IL-6 in the model group was higher than that in the control group （P<0.05）， and the expression of the above indicators was lower in each treatment group than in the model group （P<0.05）. ConclusionCalycosin can promote the proliferation and inhibit the apoptosis of astrocytes with oxidative injury by inhibiting the phosphorylation of PI3K/Akt pathway and JAK2/STAT3 pathway.
ABSTRACT
ObjectiveTo investigate the effect of calycosin-mediated glucoprotein130/Janus kinase/signal transducer and activator of transcription factor （GP130/JAK/STAT） signaling pathway on oxidative injury of astrocytes in spinal cord. MethodAstrocytes in rat spinal cord were isolated and identified by immunofluorescence detection of glial fibrillary acidic protein （GFAP）. The cells were respectively pre-treated with 5， 10， 20 μmol·L-1 calycosin for 12 h， and then 100 μmol·L-1 H2O2 （24 h） was added to induce oxidative injury. Cell counting kit-8 （CCK-8） assay was employed to detect cell proliferation and select the optimal concentration of calycosin. The following experimental groups were designed： control group， model group （100 μmol·L-1 H2O2）， calycosin group （20 μmol·L-1 calycosin）， calycosin + LY294002 group （20 μmol·L-1 calycosin + 10 μmol·L-1 LY294002）， and calycosin + Stattic group （20 μmol·L-1 calycosin + 3 μmol·L-1 Stattic）. CCK-8 assay and immunofluorescence method were used to detect the proliferation of cells and flow cytometry was applied to detect cell apoptosis and cycle. The protein expression of phosphorylated （p）-JAK2， p-STAT3， p-protein kinase B （Akt）， GP130， and interleukin-6 （IL-6） was detected by Western blotting. ResultCompared with the control group， the model group showed low proliferation activity and high apoptosis rate of cells （P<0.05）. Compared with the model group， calycosin （20 μmol·L-1） group displayed high proliferation activity and low apoptosis rate of cells （P<0.05）. Compared with calycosin （20 μmol·L-1） group， both phosphatidylinosirtol-3-kinases （PI3K） inhibitor LY294002 and STAT3 inhibitor Stattic significantly reduced the proliferation activity and increased the apoptosis rate of cells （P<0.05）. The protein expression of p-JAK2， p-STAT3， p-Akt， GP130， and IL-6 in the model group was higher than that in the control group （P<0.05）， and the expression of the above indicators was lower in each treatment group than in the model group （P<0.05）. ConclusionCalycosin can promote the proliferation and inhibit the apoptosis of astrocytes with oxidative injury by inhibiting the phosphorylation of PI3K/Akt pathway and JAK2/STAT3 pathway.
ABSTRACT
Objective To investigate the protective effect of nuclear factor E2-related factor 2(Nrf2)on hydrogen peroxide (H
ABSTRACT
Aim: To investigate the effects of Scutellarin (Scu) on microvascular endothelial cells (MEC) with ischemia/reperfusion injury (I/R) and its mechanisms involved. Methods Microvascular endothelial cells were used to build model of I/R, and the protective effects of Scu were observed. Cell viability was determined by MTT assay; SOD. MDA and LDH levels were determined by biochemical method; ICAM-1, VCAM-1, caspase-3 and LC3 mRNA expressions were detected by RT-PCR; LC3, CREB and TLR4 expressions were detected by Western blot. Results MTT assay showed that the cell viability was rescued by 50 μmol • L
ABSTRACT
@#AIM:To investigate the protective effects of naringin(Nar)phospholipid complex(NPC)on oxidative injury in retinal pigment epithelium cells(ARPE-19 cells)induced by tert-butyl hydroperoxide(t-BHP)and elucidate the underlying mechanism.<p>METHODS:The NPC was prepared by solvent method. Experimental cells are divided into seven groups: control group \〖cultured with dimethylsulfoxide(DMSO)\〗, model group(intervention with 200μmol/L t-BHP), nuclear factor erythroid 2-related factor 2(Nrf2)-siRNA group(cell transfection for Nrf2 gene), naringin group(add 200μmol/L t-BHP after pretreatment with 200μmol/L naringin medium), NPC group(add 200μmol/L t-BHP after pretreatment with 200μmol/L NPC medium), Nrf2-siRNA+ naringin group(after 200μmol/L naringin pretreatment, Nrf2 gene interference, then add 200μmol/L t-BHP)and Nrf2-siRNA+ NPC group(after 200μmol/L NPC pretreatment, Nrf2 gene interference, then add 200μmol/L t-BHP). The intracellular levels of superoxide dismutase(SOD), malondialdehyde(MDA)and total antioxidant capacity(T-AOC)were detected, intracellular level of reactive oxygen species(ROS)was detected by DCFH-DA staining method. The mRNA and protein expressions of HO-1, NQO-1, GCL and Nrf2 were detected by real-time PCR and western blot, respectively. <p>RESULTS:NPC more significantly increased the levels of SOD and T-AOC, reduced the contents of ROS and MDA than naringin in t-BHP-treated ARPE-19 cells. After naringin and NPC pre-protected ARPE-19 cells, the relative expression and protein expression of Nrf2, HO-1, NQO-1 and GCL mRNA were higher than those of the model group and Nrf2-siRNA group. There were statistically significant differences in the relative expression of 4 genes and the expression levels of 4 proteins in the naringin group and the NPC group, the Nrf2-siRNA+naringin group and the Nrf2-siRNA+NPC group. The expression of Nrf2, HO-1 and NQO-1 protein in the Nrf2-siRNA+naringin group was not significantly different than that in the Nrf2-siRNA group. Compared with the Nrf2-siRNA group, the expression of 4 proteins in the Nrf2-siRNA+NPC group was statistically significant, and the effect of NPC was significantly stronger than that of naringin.<p>CONCLUSION: After naringin forms a phospholipid complex, it can significantly increase the antioxidant capacity in cells and reduce the oxidation level. It up-regulates the expression of Nrf2 and its downstream antioxidant enzymes and phase Ⅱ detoxification enzymes by activating the Nrf2/ARE antioxidative stress pathway to better protect ARPE-19 cells from oxidative damage.
ABSTRACT
BACKGROUND: Exhaustive exercise is a vigorous physical activity performed by an organism beyond its physiological limits, and it causes a series of histological changes in the body. Myocardial exercise-induced oxidative stress injury means that the organism generates free radicals through oxidative stress signal pathway to damage myocardial cells under exhaustive exercise. OBJECTIVE: To explore the mechanism of oxidative stress injury in rat myocardium caused by one-time exhaustive exercise based on the protein kinase C (PKC)/ NOX pathway. METHODS: Thirty adult male Sprague-Dawley rats were randomly divided into a control group, an exhaustive exercise group, and an exhaustive exercise+drug group, with 10 rats in each group. The exhaustive exercise+drug group was injected with PKC inhibitor chelerythrine (5 mg/kg body weight) for 3 consecutive days. Rats in the two exercise groups exercised at a speed of 25 m/min on a 0° incline treadmill until exhaustion. Immediately after exercise, blood sample was collected from each rat, and then the rat’s left ventricle was removed for hematoxylin-eosin staining to observe the morphological changes of myocardial cells. Serum and myocardial malondialdehyde and myocardial reactive oxygen species levels were detected. The protein expressions of PKC, NOX2, NOX4 and 3-NT in rat myocardial tissue were determined by western blot. RESULTS AND CONCLUSION: The myocardial tissues in the exhaustive exercise and exhaustive exercise+drug groups were damaged, but the damage was significantly eased in the exhaustive exercise+drug group compared with the exhaustive exercise group. Compared with the control group, the reactive oxygen species level in the myocardial tissue of the exhaustive exercise group increased significantly (P < 0.05). Compared with the control group, the content of malondialdehyde in the myocardial tissues of the exhaustive exercise and exhaustive exercise+drug groups was significantly increased (P < 0.01), and the concentration of serum malondialdehyde in the exhaustive exercise group was significantly increased (P < 0.05). Compared with the control group, the expression levels of PKC, NOX2, NOX4 and 3-NT proteins in the myocardium of rats after exhaustive exercise were significantly increased (P < 0.01). Compared with the exhaustive exercise group, the expression levels of NOX2 and NOX4 in the myocardial tissue of rats significantly decreased in the exhaustive exercise+drug group (P < 0.01), and the expression of 3-NT significantly decreased (P < 0.05). Therefore, one-time exhaustive exercise can activate PKC and increase its protein expression in rat myocardial cells, which in turn induces an increase in the expression of NOX2 and NOX4 proteins in the myocardium, catalyzes the generation of large amounts of reactive oxygen species, and leads to the excessive production of peroxynitrite anions, thereby causing oxidative damage to the myocardium.
ABSTRACT
Despite the development of modern medicine, alternative medicine, which has not lost its timeliness, remains attractive for the treatment of various diseases. Glabridin, a major flavonoid of Glycyrrhiza glabra, is known for its antioxidant and anti-inflammatory activity. The aim of this study was: 1) to determine the possible protective role of glabridin against ischemia/reperfusion (I/R) injury of the intestine; 2) to evaluate the in vitrocontractile responses of ileum smooth muscles to acetylcholine after an intestinal I/R; and 3) to explain the underlying molecular mechanism of its effect. Rats were assigned to groups of six rats each; 1) I/R, 2) gla10, 3) gla20, 4) gla40, 5) N5-[imino(nitroamino)methyl]-L-ornithine, methyl ester monohydrochloride (L-NAME)+gla40, and 6) Sham group. The healing effect of glabridin was abolished by L-NAME. Glabridin did not cause contractility of the smooth muscles to acetylcholine-induced contractile responses in intestinal I/R. Yet, it increased to spontaneous basal activity.
A pesar del desarrollo de la medicina moderna, la medicina alternativa, sin perder su vigencia, sigue siendo atractiva para el tratamiento de varias enfermedades. Glabradina, el flavonoide mayoritario de Glycyrrhiza glabra, es conocido por su actividad antioxidante y antiinflamatoria. Los propósitos de este estudio fueron: 1) Determinar el posible rol protector de glabradina ante daños intestinales por isquemia/reperfusion (I/R) 2) Evaluar in vitrolas respuestas de contracción de los músculos lisos del ileum ante acetilcolina después de I/R intestinal; y 3) Explicar el mecanismo molecular subyacente de este efecto. Se asignaron grupos de seis ratas: 1) I/R, 2) gla10, 3) gla20, 4) gla40, 5) N5-[imino(nitroamino)metil]-L-ornithina, metil ester monohidrochloruro (L-NAME)+gla40, y 6) Grupo testigo. El efecto curativo de glabridina fue abolido por L-NAME. Glabridina no causó contracción en el músculo liso como respuesta acetilcolina-inducida I/R. Además, incrementa la actividad basal expontánea.
Subject(s)
Animals , Rats , Phenols/administration & dosage , Reperfusion Injury/drug therapy , Cyclic AMP/metabolism , Glycyrrhiza , Isoflavones/administration & dosage , Phenols/pharmacology , Rats, Wistar , Cyclic AMP/analysis , Cyclic GMP/metabolism , Oxidative Stress/drug effects , NG-Nitroarginine Methyl Ester , Ileum/drug effects , Ileum/chemistry , Isoflavones/pharmacology , Malondialdehyde/analysis , Muscle, Smooth/drug effectsABSTRACT
OBJECTIVE： To investigate the effect mechanism of diazoxide on the proliferation and apoptosis of chondrocytes with oxidative injury based on endoplasmic reticulum stress （ERS） pathway. METHODS： SD mice were selected for primary culture of articular chondrocytes. The 3rd generation chondrocytes were randomly divided into control group， injury model group and diazoxide group. Control group didn’t receive any treatment. The injury model group was incubated with 300 μmol/L hydrogen peroxide （H2O2） at 37 ℃ for 8 h. Diazoxide group was pretreated with 300 μmol/L diazoxide at 37 ℃ for 0.5 h，and then incubated with 300 μmol/L H2O2 for 8 h. The proliferation of chondrocytes was detected by CCK-8 assay. The apoptosis rate of chondrocytes was detected by flow cytometry. The expression of ERS-related proteins [Caspase-3， Bcl-2-associated X（Bax），C/EBP homologous protein （CHOP）] were detected by Western blotting assay. RESULTS： Compared with control group， the proliferation activity of chondrocytes in injury model group was significantly decreased， while apoptosis rate was increased significantly（P＜0.05）；the protein expression of Caspase-3， Bax and CHOP were increased significantly （P＜0.05）. Compared with injury model group， the proliferation activity of chondrocytes in diazoxide group was increased significantly， while the apoptosis rate was decreased significantly （P＜0.05）； the expression of above related proteins were decreased significantly （P＜0.05）. CONCLUSIONS： Diazoxide can improve the proliferation activity of chondrocytes with oxidative injury and inhibit their apoptosis by inhibiting ERS pathway.
ABSTRACT
Objective: To investigate the protective effect of Perillae Folium with aqueous extract (PFAE) on some key factors of Adriamycin (ADR)-induced oxidative injury in human renal tubular epithelial cells(HK-2), including the survival rate, oxidative injury indexes and cell apoptosis,in order to define the underlying mechanism. Method: A model of ADR-induced HK-2 cells oxidative injury was established in vitro, then cell viability was detected by cell counting kit-8 (CCK-8) after intervention with positive reference N-acetylcysteine (NAC) or PFAE (5,15,45 g·L-1) at different concentrations. According to the morphological changes under microscopy, the optimum concentration of PFAE was screened out for the follow-up experiments. Then, the experiments were divided into six groups:blank group, ADR (0.05 g·L-1) group, PFAE (15 g·L-1) group, ADR+PFAE (0.05+15) g·L-1 group, NAC (0.81 g·L-1) group, and ADR+NAC (0.05+81) g·L-1 group. After that, malondialdehyde (MDA), superoxide dismutase (SOD), total antioxidant capacity(TAC) were measured in the cell homogenate after 24 h administration. The level of reactive oxygen species (ROS) was detected by 2',7'-dichloroflurescin diacetate (DCFH-DA) fluorescence probe. Flow cytometry and TdT-mediated dUTP Nick-End Labeling (TUNEL) were used to monitor the cell apoptosis. Western blot was used to observed the expressions of mitochondrial apoptosis-associated proteins, like B lymphocyte tumor-2 gene (Bcl-2), Bcl-2 related X protein (Bax), cysteine aspartate protease-9 (Caspase-9), cysteine aspartate protease-3 (Caspase-3) and poly ADP-ribose polymerase (PARP), as well as their shear bodies. In addition, the phosphorylation protein expressions of p38 mitogen-activated protein kinase (p38 MAPK), extracellular signal-regulated kinase (ERK), c-Jun amino-terminal kinase (JNK) in mitogen-activated protein kinase (MAPK) signaling transduction pathway were detected by Western blot. Result: Compared with blank group, ADR group showed a decreased cell viability (PPPPPPPP-1. The ATC and SOD levels were increased in ADR+PFAE group and ADR+NAC group (PPConclusion: PFAE could alleviate the oxidative injury of HK-2 cells induced by ADR, and have an antioxidant effect, which inhibited cell apoptosis through mitochondrial apoptotic pathway and ERK/p38 MAPK signaling pathway.
ABSTRACT
AIM To explore the action mechanism of puerarin's protective effects against oxidative stress of HUVEC-12 cells induced by high glucose.METHODS HUVEC-12 cells cultured with 100 mmol/L glucose medium and 10,25,50 μmoL/L puerarin for 36 h had the cell proliferation,the levels of lactate dehydrogenase (LDH),intracellular reactive oxygen species (ROS),the activities of caspase-3,superoxide dismutase (SOD)and catalase (CAT),and the contents of malondialdehyde (MDA) and glutathione (GSH) measured.The mRNA expressions of SIRT1 and PGC-1α were detected by real-time flu orescence quantitative PCR,and the protein contents of SIRT1 and PGC-1 α were determined by enzyme-linked immunosorbent assay.RESULTS The puerarin treatment to HUVEC-12 cells resulted in markedly lowered LDH level,caspase-3 activity,intracellular levels of MDA and ROS,and notable improvement of the cell viability,the activities of SOD and CAT,GSH content,the mRNA expressions and the protein contents of SIRT1 and PGC-1α as well.CONCLUSION The protective effect of puerarin on high glucose-induced oxidative damage of HUVEC-12 cells may be attributed to the SIRT1/PGC-1α pathway activation.
ABSTRACT
To screen the toxic polar fractions of Daphne genkwa, compare the toxicity of D. genkwa on crypts epithelial cells IEC-6 before and after vinegar processing, and preliminarily investigate the mechanism of D. genkwa vinegar processing on toxicity reducing. The proliferation of IEC-6 cells was observed by MTT. The levels of superoxide dismutase (SOD), malondialdehyde (MDA), glutathione (GSH), lactate dehydrogenase (LDH), as well as the enzyme activity of Na⁺-K⁺-ATPase and Ca²⁺-Mg²⁺-ATPase were determined in IEC-6 cells to evaluate the oxidative damages degree of IEC-6 cells. The apoptosis and cell cycle were analyzed by Flow Cytometry. The results showed that the dichloromethane extraction was the toxic polar fraction of D. genkwa, and after vinegar processing, the toxicity of dichloromethane fraction was significantly reduced (<0.01). As compared with the blank control group, the dichloromethane fraction of D. genkwa can obviously decrease the levels of SOD, Na⁺-K⁺-ATPase, Ca²⁺-Mg²⁺-ATPase (<0.01) and content of GSH, but increase the level of LDH and MDA in cell supernatant (<0.01). Besides, it obviously increased the early and late apoptotic rate of IEC-6 cells, obviously decreased the proportion of G₁stage cells, increased the ratio of S stage cells and M stage cells (<0.01). After vinegar processing, as compared with D. genkwa groups of various doses, it can significantly increase the levels of SOD, Na⁺-K⁺-ATPase, Ca²⁺-Mg²⁺-ATPase (<0.01) and content of GSH, decrease the level of LDH, MDA(<0.01), significantly decrease the early and late apoptosis rate of IEC-6 cells (<0.01), increase the proportion of G₁stage cells, and decrease the ratio of S stage cells and M stage cells (<0.01). Vinegar processing can reduce the toxicity of dichloromethane fraction of D. genkwa, and its mechanism may be associated with improving the activity of antioxidant enzymes and permeability in IEC-6 cells, and decreasing the oxidative damage.
ABSTRACT
OBJECTIVE: To investigate the effects of Silibinin capsules combined with routine therapy on serum oxidative injury and liver function of patients with alcoholic liver disease (ALD) complicated with early liver fibrosis.METHODS: The patients with ALD complicated with early liver fibrosis selected from our hospital during Apr. 2013-Jan. 2015 were divided into control group and study group according to random number table, with 38 cases in each group. Control group received routine symptomatic treatment [temperance, oral administration of Inosine tablets (0. 4 g/times, bid), vitamin and microelement supplementation, etc. ]. Study group was additionally given Silibinin capsule (70 mg/time, bid) orally, for consecutive 48 weeks, on the basis of control group. The clinical efficacies and the incidence of ADR were compared between 2 groups. The changes of serum oxidative injury indexes (SOD, MDA), liver fibrosis indexes [laminin (LN), hyaluronic acid (HA), type IV collagen (IV-C), type Ⅲ procollagen (PC Ⅲ)] and liver function indexes (ALT, AST) were recorded before and after treatment. RESULTS: Before treatment, there was no statistical significance in baseline information between 2 groups (P>0. 05). After treatment, total response rate of study was significantly higher than that of control group (P<0. 05), but there was no statistical significance in the incidence of ADR between 2 groups (P>0. 05). Compared with before treatment, serum levels of MDA, liver fibrosis indexes and liver function indexes in 2 groups were decreased significantly (P<0. 05), while SOD levels were increased significantly (P<0. 05); the improvement of study group was more significant than control group (P<0. 05). CONCLUSIONS: Silibinin capsules combined with routine treatment can enhance clinical efficacy of ALD patients with early liver fibrosis, mainly manifesting as improving oxidative stress, regulating liver function and inhibiting the development of liver fibrosis.
ABSTRACT
OBJECTIVE: To observe the effects of maternal low-level decabromodiphenyl( BDE209) exposure on nervous system and secretion of thyroid hormones in offspring mice. METHODS: Sixty-four specific pathogen free female,aged 4 weeks Kunming mice were used. These mice were randomly divided into control group and low,medium and high exposure groups after successful mating was confirmed. The rats of control group were fed with 0. 01 L/kg body mass of peanut oil.The maternal mice in the experimental groups were given BDE209 at doses of 50,100 and 300 μg/kg body mass by oral gavage once per day. Continuous exposure was given until 21 days after birth of offspring,the exposure model from gestation to lactation was established. At the end of the exposure,10 mice of each group including half female and half male were randomly selected and the body mass and growth development status were observed. Morris water maze was used to examine the learning and memory ability in offspring mice. The serum levels of total triiodothyronine( TT3),free triiodothyronine( FT3), total tetraiodothyronine( TT4) and free tetraiodothyronine( FT4) were measured by chemiluminescence immunoassay. The levels of glutathione transferase( GST), superoxide dismutase( SOD) and malondialdehyde( MDA) level in hippocampus were measured by colorimetry. RESULTS: The escape latency of the medium exposure group was longer than that of the control group( P < 0. 05). The escape latency of high exposure group was longer than that of control group,low exposure group and medium exposure group( P < 0. 05). The time of quadrant movement and number of crossing the platform in offspring rats in high exposure group were less than that of the control group and the low and medium exposure groups( P < 0. 05). The serum levels of TT3,FT3,TT4 and FT4 increased,the activities of GST and SOD in hippocampus tissue decreased,the MDA level increased with the increasing exposure dose( P < 0. 05).CONCLUSION: Maternal low-level BDE209 exposure can result in decrease the learning and memory ability of offspring mice. It also can increase the serum thyroid hormone level and induce oxidative stress injury in hippocampus in a dose dependent manner in offspring mice.
ABSTRACT
In the present study, three new triterpenoids, 23-hydroxyurs-12, 18-dien-28-oic acid 3β-O-α-L-arabinopyranoside (1), 23-hydroxyurs-12, 18-dien-28-oic acid 3β-O-β-D-glucuronopyranoside-6-O-methyl ester (2), and urs-12, 18-dien-28-oic acid 3β-O-β-D-glucuronopyranoside-6-O-methyl ester (3), and a known triterpenoid, 3β-hydroxy-urs-2, 18-dien-28-oic acid (4, randialic acid B), were isolated from the aerial parts of Ilex cornuta. Their structures were identified by the spectroscopic analyses (IR, ESI-MS, HR-ESI-MS, and 1D and 2D NMR) and chemical reactions. Compound 4 showed significant cell-protective effects against HO-induced H9c2 cardiomyocyte injury. Compounds 1-4 did not show any significant DPPH radical scavenging activity.
Subject(s)
Animals , Rats , Biphenyl Compounds , Metabolism , Cardiovascular Agents , Chemistry , Pharmacology , Hydrogen Peroxide , Metabolism , Ilex , Chemistry , Molecular Structure , Myocardium , Cell Biology , Pathology , Myocytes, Cardiac , Picrates , Metabolism , Plant Components, Aerial , Chemistry , Plant Extracts , Chemistry , Pharmacology , Triterpenes , Chemistry , PharmacologyABSTRACT
Objective To investigate the role of leukemia inhibitory factor (LIF) in cell survival of cone photoreceptors and the underlying mechanism following oxidant injury.Methods 661W cells were cultured and randomly divided into normal control group,LIF intervention group,H2O2 intervention group,S3 I201 intervention group,H2O2 + LIF intervention group,H2O2 + S3I201 intervention group and H2O2 + LIF + S3I201 intervention group,according to the different intervention drugs.MTT assay and Western blot were used to detect the effects of LIF pretreatment on 661W activity and the expression of STAT3 protein and its phosphorylation level after oxidative damage.Using STAT3-specific inhibitor S3I201 to block the STAT3 signal transduction pathway,MTT assay and real-time PCR were used to detect the effects of STAT3 signaling pathway on 661 W activity and the mRNA expression of survivin bcl-2 and bcl-xi after oxidative damage.Results Compared to H2O2 intervention group,the relative protein expression of p-Tyr705-STAT3 was increased in H2O2 + LIF intervention group,the difference was statistically significant (P ＜ O.05);The relative protein expression of p-Tyr705-STAT3 was decreased in H2O2 + S3I201 intervention group,the difference was statistically significant (P ＜ 0.05).Compared to H2O2 intervention group,the cell activity of 661W cells was increased in H2O2 + LIF intervention group,the difference was statistically significant(P ＜0.05).Compared to H2O2 + LIF intervention group,the cell activity of 661W cells was decreased in H2O2 + LIF + S3I201 intervention group,the difference was statistically significant (P ＜ 0.05).Compared to normal control group,the mRNA expression of bcl-2 and bcl-xl were increased in LIF intervention group,H2O2 intervention group and H2O2 + LIF intervention group,the differences were statistically significant (all P ＜ 0.05).Compared to H2O2 + LIF intervention group,the mRNA expression of bcl-2 and bcl-xl were decreased in H2O2 + LIF + S3I201 intervention group,the differences were statistically significant (all P ＜ 0.05).Conclusion LIF has protective effect on oxidative injury of cone photoreceptors,and it may contribute to cell survival through activation of the STAT3 signaling transduction pathway and its related survivin.
ABSTRACT
Objective To explore the effect of D-galactose(D-gal) on murine pancreatic injury and its pathogenesis.Methods C57BL/6J mice were randomly divided into control group and D-gal model group [D-gal 120 mg/(kg · d) for 42 days].On the 2nd day after drug injection completed,the peripheral blood was taken for measuring the level of fasting blood glucose(FBG) and fasting insulin(FINS);and then the organ index of pancreas was calculated by the ratio of pancreatic wet weight(mg) and mouse body weight(g);HE stain was routinely prepared to observe the histologic structure of pancreatic tissue;the TEM was used to analyze ultrastructural changes of pancreatic cells;the pancreatic frozen sections were prepared to test senescence-associated β-galactosidase (SA-β-gal) and its relative absorbance(RA) of positively stained cells in the pancreatic islets;immunohistochemistry assays to study advanced glycation end products (AGEs) and its RA;pancreas tissue homogenate was made to detect the content of superoxide dismutase(SOD),malonaldehyde(MDA) and total antioxidant capacity(T-AOC).Results In D-gal group mice,the FBG increased(P＜0.05) and FINS reduced;pancreas wet weight and organ increased obviously (P＜0.01);light microscopic structure of the pancreas presented without typical pathologic change,however the single nucleated cell's area within the islet was increased significantly(P＜0.05);the pancreas endocrine and exocrine cells were showed the ultrastructure damaged and lipofuscin formation increased;the RA of positive pancreas cells in SA-β-gal staining increased(P＜0.05);the RA of AGEs positive regional expression markedly increased (P＜0.01);the content of SOD and T-AOC decreased (P ＜ 0.05),the content of MDA increased (P ＜ 0.01).Conclusions Aging mice model replicated by D-gal can cause the pancreatic injury,its mechanisms may be closely related to oxidative injury of pancreatic cells caused by D-gal.
ABSTRACT
Objective To evaluate effects of ascorbic acid on proliferative activity of cultured melanocytes in vitro, as well as on H2O2?induced oxidative injury in melanocytes. Methods The optimal concentration of ascorbic acid solution and median lethal dose of H2O2 solution were determined by CCK?8 assay for the following experiment. Cultured melanocytes were classified into the control group, ascorbic acid group, H2O2 group and combination group. During the first 24 hours, the control group and H2O2 group were treated with M254 medium, while the ascorbic acid group and combination group with ascorbic acid solution. During an additional 24?hour period, the control group and ascorbic acid group were treated with M254 medium, while the H2O2 group and combination group with H2O2 solution at the median lethal dose. After 48?hour treatment, CCK?8 assay and flow cytometry were performed to determine the survival rate and apoptosis rate of melanocytes, respectively, in the 4 groups. Biochemical methods were used to evaluate the superoxide dismutase(SOD)activity and determine the malondialdehyde(MDA)concentration, and fluores?cent staining was conducted to detect the level of reactive oxygen species(ROS)in the control group, H2O2 group and combination group. Results The optimal concentration of ascorbic acid solution was 1 000μmol/L, and the median lethal dose of H2O2 solution was 300 μmol/L. The cell survival rate, apoptosis rate, SOD activity, MDA concentration and ROS fluorescence intensity in the control group were 100% ± 4.99%, 6.90% ± 0.87%, 54.71 ± 4.75 U/mgprot, 263.39 ± 20.17 nmol/mgprot and 342.16 ± 27.36 respectively. Compared with the control group, H2O2 solution could significantly increase the cell apoptosis rate(16.47%± 1.07%), SOD activity(103.62 ± 10.44 U/mgprot), MDA concentration(493.70 ± 31.36 nmol/mgprot)and intracellular ROS fluorescence intensity (782.48 ± 36.25), but decrease the survival rate of melanocytes (39.07% ± 2.94%), while ascorbic acid solution markedly down?regulated the H2O2?induced apoptosis (11.83%± 0.95%), SOD activity(76.73 ± 5.20 U/mgprot), MDA concentration(371.82 ± 23.05 nmol/mgprot) and ROS level (475.64 ± 52.18), but increased the cell survival rate (74.31% ± 5.53%). Conclusion Ascorbic acid solution at the concentration of 1 000 μmol/L can not only promote proliferative activity of melanocytes, but also protect melanocytes from H2O2?induced oxidative injury.
ABSTRACT
Objective To investigate the alteration of energy metabolism and oxidative injury in the myocardia suffering from lethal ventricular tachyarrhythmia (LVTA). Methods Two LVTA-SCD SD rat models, induced by aconitine injection or coronary artery ligation (CAL), respectively, were developed. Rats that died from over-anaesthesia or CAL-induced heart failure were served as their controls, respectively. Mitochondrial membrane potential (MMP), reactive oxygen species (ROS), malonaldehyde (MDA), phosphocreatine (PCr) in the ventricular myocardia, and serum troponin I (cTnI) were detected, and compared between LVTA rats and their controls. Results Fourteen ACO-LVTA and six CAL-LVTA rats were successfully developed. As compared to their controls, ACO-LVTA and CAL-LVTA rats had higher ROS and MDA content, and lower concentration of PCr in the ventricular myocardia. MDA content in ACO-LVTA group is signiifcantly higher than that of its control (P<0.05). MMP in myocardia of ACO-LVTA is lower than that of its control, but is higher than those of two CAL groups. Serum cTnI in rats of both LVTA models is higher than those of their controls and pre-treated control. Specially, serum cTnI in CAL-LVTA was signiifcantly higher than that of ACO-LVTA and its control (P<0.01). The myocardial ROS content is correlated with the duration of VT and VF (P<0.05), with correlation coefifcients being 0.44 and 0.46, respectively. Conclusions After LVTA, the ventricular myocardia had lower MMP and PCr content, higher concentration of ROS, MDA, as well as higher serum cTnI than their controls, indicative of oxidative injury and alteration of energy metabolism under LVTA-SCD.