MnSOD mRNA Expression Research Articles

Reproductive toxicity of hexabromocyclododecane in rotifer Brachionus plicatilis: Involvement of reactive oxygen species and calcium signaling pathways

To assess the toxicity of Hexabromocyclododecane (HBCD), the population, individual, and cellular biochemical parameters of the rotifer Brachionus plicatilis exposed to different concentrations of HBCD were investigated. The results showed that the population growth rate, reproductive period, and offspring number in B. plicatilis significantly decreased under 324 μg/L and 648 μg/L HBCD. Antioxidant enzyme activity and mRNA expression of CAT and Mn-SOD were promoted at low concentrations (32 μg/L and 64 μg/L) and inhibited at high concentrations (324 μg/L and 648 μg/L), while MDA content accumulated continuously with increasing HBCD concentrations, indicating that HBCD induced oxidation imbalance in rotifers. Further evidence was provided by the correlation between DNA fragmentation and physiological changes. The increased intercellular concentration of Ca2+ and the expression of CaM mRNA suggested that HBCD activated pathways related to calcium signaling. In summary, the excessive production of ROS induced by HBCD was considered to be the main cause of reproductive toxicity.

Berberine Attenuates Acetamiprid Exposure-Induced Mitochondrial Dysfunction and Apoptosis in Rats via Regulating the Antioxidant Defense System.

Acetamiprid (ACMP) is a neonicotinoid insecticide that poses a significant threat to the environment and mankind. Oxidative stress and mitochondrial dysfunction are considered prime contributors to ACMP-induced toxic effects. Meanwhile, berberine (BBR) a natural plant alkaloid, is a topic of interest because of its therapeutic and prophylactic actions. Therefore, this study evaluated the effects of BBR on ACMP-mediated alterations in mitochondrial functions and apoptosis in rat liver tissue. Male Wistar rats were divided into four groups: (I) control, (II) BBR-treated, (III) ACMP-exposed, and (IV) BBR+ACMP co-treated groups. The doses of BBR (150 mg/kg b.wt) and ACMP (1/10 of LD50, i.e., 21.7 mg/kg b.wt) were given intragastrically for 21 consecutive days. The results showed that the administration of ACMP diminished mitochondrial complex activity, downregulated complex I (ND1 and ND2) and complex IV (COX1 and COX4) subunit mRNA expression, depleted the antioxidant defense system, and induced apoptosis in rat liver. BBR pre-treatment significantly attenuated ACMP-induced mitochondrial dysfunction by maintaining mitochondrial complex activity and upregulating ND1, ND2, COX1, and COX4 mRNA expression. BBR reversed ACMP-mediated apoptosis by diminishing Bax and caspase-3 and increasing the Bcl-2 protein level. BBR also improved the mitochondrial antioxidant defense system by upregulating mRNA expression of PGC-1α, MnSOD, and UCP-2 in rat liver tissue. This study is the first to evaluate the protective potential of BBR against pesticide-induced mitochondrial dysfunction in liver tissue. In conclusion, BBR offers protection against ACMP-induced impairment in mitochondrial functions by maintaining the antioxidant level and modulating the apoptotic cascade.

Lead induced genotoxicity and hepatotoxicity in zebrafish (Danio rerio) at environmentally relevant concentration: Nrf2-Keap1 regulated stress response and expression of biomarker genes

Genotoxic and hepatotoxic potentials of Pb at an environmentally relevant concentration (5 ppm) in zebrafish were investigated in the present study. Erythrocytic nuclear abnormality tests revealed the increased frequencies of abnormal erythrocytes after Pb exposure, indicating a strong genotoxic potential of Pb. Multiple stress-related parameters were further evaluated in liver, the major detoxifying organ. Pb caused increased production of ROS, which in turn caused severe oxidative stress. As a result, lipid peroxidation was increased, whereas reduced glutathione level and catalase activity was decreased. Alterations in liver histoarchitecture also served as evidence of Pb-induced hepatotoxicity. Pb-induced ROS stress triggered upregulation of Nrf2, Nqo1, Ho1; downregulation of Keap1, and altered mRNA expressions of Mn-sod, Cu/Zn-sod, gpx1, cyp1a, ucp2 suggesting involvement of Nrf2-Keap1-ARE signaling in cellular defence. Nrf2-keap1 is a sensitive biomarker of Pb-induced ROS stress. Overexpression of Hsp70 and other genes in hepatocytes might help cell survival under oxidative stress generation.

Andrographolide Reverses Doxorubicin Resistance in Human Breast Cancer Stem Cells by Regulating Apoptotic Gene Expressions

BACKGROUND: Breast cancer stem cells (BCSCs) have been identified as playing a crucial role in therapeutic resistance. This resistance can be attributed to the anti-apoptotic protein survivin and the antioxidant MnSOD high expression. To overcome the resistance to doxorubicin (DOX), this study proposed the utilization of andrographolide (ANDRO), the primary bioactive compound in Andrographis paniculata leaves. The objective was to examine the role of andrographolide in regulating survivin, caspase-9, and caspase-3 gene expressions to reverse doxorubicin resistance in human BCSCs.METHODS: BCSCs were exposed to 0.1 µM DOX every two days or 50 µM rotenone (ROT) for 6 hours, subsequently supplemented with 0.3 mM ANDRO. Superoxide and peroxide levels were measured using DHE and DCFH-DA assay. The MnSOD, survivin, caspase-9, and caspase-3 mRNA expression levels were analyzed using qRT-PCR. Protein expressions were evaluated using Western blotting assay. MnSOD activity was determined using xanthine oxidase inhibition assay. The apoptotic cells were determined using Annexin-V/PI staining.RESULTS: This study indicated that the cytotoxic mechanisms of DOX, similar to ROT, in BCSCs were attributed to oxidative stress, as evidenced by an elevation in superoxide rather than peroxide levels, accompanied by a decrease in MnSOD activity. This study also highlighted that ANDRO reversed DOX resistance in BCSCs subjected to repeated DOX treatment by downregulating survivin and upregulating caspase-9 and caspase-3 mRNA expressions, thereby activating the intrinsic apoptotic pathway.CONCLUSION: This study provides insights into the role of ANDRO in modulating the expression of apoptotic genes, such as survivin, caspase-9, and caspase-3, to overcome DOX resistance in BCSCs.KEYWORDS: breast cancer, breast cancer stem cell, andrographolide, doxorubicin, oxidative stress, apoptosis

Growth performance, physiological and antioxidant capacity responses to dietary fish meal replacement with insect meals for aquaculture: A case study in red claw crayfish ( Cherax quadricarinatus )

A 60-day feeding trial was conducted to evaluate the effects of yellow mealworm (YM) (Tenebrio molitor) (9%, 18%, 27% and 36%) and black soldier fly larvae (BSFL) (Hermetia illucens) (7%, 14%, 21% and 28%) meal replacing fish meal (FM) on growth performance and antioxidant capacity of red claw crayfish (Cherax quadricarinatus). Nine diets, including control, were fed with quadruplicate of 20 crayfish (0.20 ± 0.05 g). The highest weight gain (WG) and lowest feed conversion ratio were observed in crayfish fed YM18 and BSFL14. The highest superoxide dismutase activity and the lowest malondialdehyde content were detected in the hepatopancreas of the crayfish fed YM18 and BSFL14. Compared with the control group, crayfish fed BSFL14 had higher levels of glutathione peroxidase, catalase acid and alkaline phosphatase and the lowest malondialdehyde (MDA) content in hemocytes. The mRNA expression of CqBAF, HSP70 and MnSOD in the hepatopancreas of crayfish showed an increasing trend. Based on the second-order polynomial regression analysis of WG, the optimal dietary replacement level of FM with YM meal and BSFL was determined to be 21.9% and 17.1% for juvenile C. quadricarinatus.

The Impact of Background-Level Carboxylated Single-Walled Carbon Nanotubes (SWCNTs−COOH) on Induced Toxicity in Caenorhabditis elegans and Human Cells

Single-walled carbon nanotubes (SWCNTs) are widely utilized for industrial, biomedical, and environmental purposes. The toxicity of Carboxylated SWCNTs (SWCNTs−COOH) in in vivo models, particularly Caenorhabditis elegans (C. elegans), and in vitro human cells is still unclear. In this study, C. elegans was used to study the effects of SWCNTs−COOH on lethality, lifespan, growth, reproduction, locomotion, reactive oxygen species (ROS) generation, and the antioxidant system. Our data show that exposure to ≥1 μg·L−1 SWCNTs−COOH could induce toxicity in nematodes that affects lifespan, growth, reproduction, and locomotion behavior. Moreover, the exposure of nematodes to SWCNTs−COOH induced ROS generation and the alteration of antioxidant gene expression. SWCNTs−COOH induced nanotoxic effects at low dose of 0.100 or 1.00 μg·L−1, particularly for the expression of antioxidants (SOD-3, CTL-2 and CYP-35A2). Similar nanotoxic effects were found in human cells. A low dose of SWCNTs−COOH induced ROS generation and increased the expression of catalase, MnSOD, CuZnSOD, and SOD-2 mRNA but decreased the expression of GPX-2 and GPX-3 mRNA in human monocytes. These findings reveal that background-level SWCNTs−COOH exerts obvious adverse effects, and C. elegans is a sensitive in vivo model that can be used for the biological evaluation of the toxicity of nanomaterials.

Effect of medical herbs in Tian-Dong-Tang-Gan powder on the oxidative stress induced by ammonia and nitrite in Litopenaeus vannamei

In order to evaluate the effect of different doses of Tian-Dong-Tang-Gan powder (TDTGP) on the immune function of Litopenaeus vannamei, and the molecular mechanism of anti-ammonia stress and anti-nitrite stress, and screening for the best dose, a 35-day feeding experiment and a 72-h ammonia stress or nitrite stress experiment were carried out. The results showed that when compared with the blank control group, the different doses of TDTGP could increase the body weight (BW), percent weight gain (PWG), body length (BL) and percent body length growth (PBLG), and the activities of acid phosphatase (ACP), phenol oxidase (PO), inducible nitric oxide synthase (i-NOS) and the total antioxidant capacity (T-AOC) in the hemolymph of L. vannamei during the feeding experiment, and the effect of TDTGP-4 treatment group was the best. When exposed ammonia or nitrite stress for 72 h, the PO activity was decreased in the hemolymphs, and the mRNA expression of PPO, MT, GSH-Px, Mn-SOD, HSP70 and HIF-1α in the hepatopancreas of L. vannamei. However, different TDTGP doses were fed to the shrimp followed by the application of ammonia and nitrite stresses. The activities of ACP, PO, i-NOS and T-AOC in the hemolymphs remained stable, and the mRNA expression of PPO, ACP, metallothionein (MT), glutathione peroxidase (GSH-Px), superoxide dismutase (Mn-SOD), heat shock protein 70 (HSP70), Hypoxia inducible factor 1α (HIF-1α) and other factors in the hepatopancreas could be maintained or improved to varying degrees in comparison with the blank control group and positive group. These results indicated that the TDTGP can improve growth performance and non-specific immune response of L. vannamei, and by maintaining the mRNA expression of the antioxidative stress factor, such as MT, GSH-PX, Mn-SOD, HSP70, HIF-1α and PPO and ACP in the hepatopancreas, acting against the damage of hepatopancreas caused by ammonia and nitrite stress in L. vannamei. The changes in BW, WG, activities of PO and ACP, and mRNA expression levels of PO, Mn-SOD and ACP indicated that the optimum supplement of dietary TDTGP should be 4 g per·kg.

Rosemary extract reverses oxidative stress through activation of Nrf2 signaling pathway in hamsters fed on high fat diet and HepG2 cells

• Diet rosemary extract (RE) alleviated high-fat induced oxidative damage in vivo and in vitro. • Diet RE enhanced the antioxidant enzyme activity, gene and protein expression level. • The protective effect of diet RE was achieved by regulating Nrf2 pathway. Rosemary extract (RE), a useful component isolated from Rosmarinus officinalis L. , was recently reported to protect cells against the harmful effects of ROS. However, the underlying mechanisms of this action remained unknown. We investigated the defensive effect of RE on the oxidative stress injury resulted from high fat diet in hamsters and oleic acid (OA) in HepG2 cells respectively, and its underlying mechanisms. Treatment with RE alleviated liver steatosis and reduced plasma and liver content of TG and TC, as well as plasma content of AST and ALT. RE apparently increased the activity of SOD, GSH-Px, CAT and decreased MDA content. RE notably increased CuZn-SOD, Mn-SOD, CAT, GSH-Px, GCLC, GCLM and Nrf2 mRNA and protein expression level. Nrf2 shRNA knockdown in HepG2 cells increased OA-induced ROS generation, and effectively altered OA-induced cytotoxicity, suggesting that Nrf2 is a crucial mediator against high-fat induced oxidative damage.

Lactobacillus plantarum KFY02 enhances the prevention of CCl4-induced liver injury by transforming geniposide into genipin to increase the antioxidant capacity of mice

This study investigated Lactobacillus plantarum KFY02 (KFY02) and its ability to enhance the antioxidant capacity of the body by converting geniposide to genipin, thereby decreasing the liver damage induced by CCl4 in mice. The experimental results showed that geniposide is partially converted into genipin by KFY02. In liver-injured mice, KFY02 and geniposide could decrease the liver index, the AST, ALT, TG, and MDA serum levels, and the levels of cytokines IL-6, IL-12, TNF-α, and IFN-γ while up-regulating SOD and GSH-Px levels. KFY02 and geniposide increased the mRNA and protein expression of Mn-SOD, Cu/Zn-SOD, CAT, GSH-Px, and IκB-α and decreased the mRNA and protein expression of COX-2, iNOS, and NF-κB in the liver tissue of liver-injured mice. In summary, the conversion of geniposide to genipin by L. plantarum KFY02 can effectively enhance the body's antioxidant capacity to prevent CCl4-induced liver damage in mice.

Antioxidative Potential of Red Deer Embryos Depends on Reproductive Stage of Hind as a Oocyte Donor

Simple SummaryDeer breeding tends to select animals for obtain the high meat quality and in case of males preferred shape and weight of antlers. Fertilization in vitro (IVF) using high-indexing parents results favorable features. Moreover, evaluation of effective method of IVF on Cervus elaphus as a model, will be useful for application on Cervids in danger of extinction. The effectivity of IVF depends on quality of gametes and proper development of embryo. The aim was to compare the blastocyst stages of red deer embryos in respect of IVF efficiency, morphology, apoptotic and proliferative abilities, and antioxidative potential according to the reproductive status of hinds. We used three experimental groups, including the ovaries collected post mortem on the 4th and 13th days of the estrous cycle (farmed animals) and during pregnancy (wild animals). Frozen-thawed epididymal semen was used for IVF. Blastocyst quality, apoptotic, and antioxidative potential of blastocysts were evaluated. Results indicate that red deer embryos on blastocyst stage received in vitro collected from hinds on 4th day of the estrous cycle as an oocyte donor are characterized by better antioxidative potential and qualities to those developed from oocytes collected from hinds on 13th day of the estrous cycle and pregnancy.The aim was to compare the blastocyst stages of red deer embryos in respect of in vitro fertilization (IVF) efficiency, morphology, apoptotic and proliferative abilities, and antioxidative potential according to the reproductive status of hinds. We used three experimental groups, including the ovaries collected post mortem on the 4th and 13th days of the estrous cycle and during pregnancy (n = 18). After oocyte maturation, frozen-thawed epididymal semen was used for IVF. Blastocyst quality, apoptotic potential by determining the mRNA expression of BAX, BCL-2, OCT4, SOX2, and placenta-specific 8 gene (PLAC8), and antioxidative potential of blastocysts were evaluated by determining the mRNA expression of CuSOD, MnSOD, and GPX as well as the enzymatic activity of superoxide dismutase and reduced glutathione. The highest development rate of expanded blastocyst, mRNA expression of BCL-2, OCT4, SOX2, and PLAC8 and mRNA expression and enzymatic activity of the antioxidative factors increased (p < 0.05) in blastocysts developed from the oocytes collected on the 4th day, compared to those developed from the oocytes collected on the 13th day of the cycle and during pregnancy. Our study indicates that the 4th day of the estrous cycle is the most effective period for oocyte collection for IVF and embryo development in hinds, considering quality parameters and antioxidative potential of the blastocysts.

Ganoderma lucidum spore oil (GLSO), a novel antioxidant, extends the average life span in Drosophila melanogaster

In ancient China, Ganoderma lucidum was believed to be a medical fungus that could increase lifespan. Recently, pharmacologic studies have found that polysaccharide peptides and triterpenoids extracted from Ganoderma lucidum have various physiological effects as active compounds. However, the effects of spore oil isolated from Ganoderma lucidum remains unknown. In this study, the biological effects of Ganoderma lucidum spore oil (GLSO) were evaluated using a Drosophila melanogaster model. Compared with untreated groups, groups treated with GLSO had significantly longer average and maximum lifespan in both normal conditions and under oxidative stress. The activities of various antioxidant enzymes were measured to determine the antioxidant effect of GLSO. GLSO treatment markedly enhanced total superoxide dismutase (SOD) and catalase (CAT) activity and decreased levels of malondialdehyde (MDA). Further, we found dose-dependent increases in the mRNA expression of Cu, Zn-SOD, Mn-SOD, and CAT in GLSO-treated groups. These results suggest that GLSO may effectively eliminate free radicals and extend lifespan in Drosophila. Future work should investigate the value of GLSO as a functional food for the prevention of aging in larger animal models.

Modulation of glutathione S-transferase and superoxide dismutase in response to heavy metals in brackish water flea Diaphanosoma celebensis

Superoxide dismutase (SOD) and glutathione S-transferase (GST) serve as biomarkers of oxidative stress in aquatic organisms. However, the modulatory effects of heavy metals on the expression of small crustaceans are yet unknown. The acute toxicity of inorganic arsenic (As), cadmium (Cd), and lead (Pb) to the brackish water flea Diaphanosoma celebensis (4 days old) was evaluated to determine their sublethal concentrations. Modulations in the activities and transcription of SODs (Cu/Zn-SOD and Mn-SOD) and GSTs (mu, kappa, theta, and sigma) in response to sublethal concentrations of heavy metals were investigated for 48 h. The 48-h median lethal concentration (LC50) was determined to be 2.62 mg/L, 2.60 mg/L, and 0.37 mg/L for Cd, As, and Pb, respectively, and the order of toxicity in this species was Pb > Cd = As. SOD and GST activities were induced after exposure to all metals; in particular, GST activity was sensitively enhanced upon As exposure. Real-time reverse-transcriptase polymerase chain reaction showed that GST mu and GST sigma were specific for As, and GST kappa and GST theta for Cd. Mn-SOD mRNA expression was more sensitive to As. These findings suggest that D. celebensis SOD and GST may be involved in the antioxidant defense mechanisms to heavy metals. These genes may serve as useful biomarkers for risk assessment of heavy metal pollution in aquatic ecosystems. This study may be helpful for the better understanding of the molecular mode of action of heavy metals in small crustaceans and provide preliminary data for future studies evaluating the combined effects of heavy metal mixture.

Effects of light-emitting diodes on thermally-induced oxidative stress in the bay scallop Argopecten irradians

ABSTRACTWater temperature is an important stressor that affects the physiological and biochemical responses of scallops. In this study, we investigated the effect of different light-emitting diodes (LEDs; red, green and blue) on oxidative stress in Argopecten irradians. PCR revealed MnSOD mRNA expression in the digestive diverticula, gill, adductor muscle and eye. CAT and HSP70 mRNA were expressed in the digestive diverticula, gill and adductor muscle. Additionally, we measured the changes in the expression of HSP70, MnSOD and CAT as well as H2O2 levels during thermal/laboratory stress. In the digestive diverticula, gill and adductor muscle, the mRNA expressions and activities and H2O2 levels significantly increased in response to thermal changes. The gene expressions and activities and H2O2 levels were significantly lower in scallops that received green LED light than in those that received no mitigating treatment. A comet assay revealed that thermal change groups had increased rates of nuclear DNA damage; however, treatment with green LED reduced the frequency of damage. The results indicated that low or high water temperature conditions induced oxidative stress in A. irradians but that green LED significantly reduced this stress.

Preventive effect of lemon seed flavonoids on carbon tetrachloride-induced liver injury in mice.

The aim of this study was to determine the preventive effect of lemon seed flavonoids (LSF) on carbon tetrachloride-induced liver injury in mice. Liver injury was induced by injection with 2 mL kg−1 of carbon tetrachloride after administration of LSF by gavage. Liver index, serological parameters, and expression intensities of related mRNA and protein in the liver tissue were observed. The results indicated that LSF reduced liver weight and liver index, downregulated serum levels of AST, ALT, ALP, TG, TC, BUN, NO, and MDA, and upregulated levels of ALB, SOD, CAT, and GSH-Px in the mice with liver injury. It also downregulated serum cytokines, such as IL-6, IL-12, TNF-α, and IFN-γ in these mice. qPCR and western blot confirmed that LSF upregulated mRNA and protein expression of Mn-SOD, Cu/Zn-SOD, CAT, GSH-Px, and IκB-α, and downregulated expression of NF-κB-p65, iNOS, COX-2, TNF-α, IL-1β, and IL-6 in the liver tissue of mice with liver injury. The preventive effect on carbon tetrachloride-induced liver injury was attributed to (−)-epigallocatechin, caffeic acid, (−)-epicatechin, vitexin, quercetin, and hesperidin, which were active substances that were detected in LSF by HPLC. Moreover, the effect of LSF is similar to that of silymarin, but the synergistic effect of the five active substances working in concert acted to produce a more robust liver-protecting effect.

Profiling of Gene Expression Associated with Stemness and Aggressiveness of ALDH1A1-Expressing Human Breast Cancer Cells.

BackgroundIt has been widely reported that breast cancer aggressiveness may be driven by breast cancer stem cells (BCSCs). BCSCs display stemness properties that include self-renewal, tumourigenicity and pluripotency. The regulation of gene expression may have important roles in BCSC stemness and aggressiveness. Thus, the aim of this study was to examine the stemness and aggressiveness gene expression profile of BCSCs compared to MCF-7 and MDA-MB-231 breast cancer cells.MethodsHuman ALDH1+ BCSCs were grown in serum-free Dulbecco’s Modified Eagle Medium (DMEM)/F12, while MCF-7 and MDA-MB-231 were cultured in DMEM supplemented with 10% foetal bovine serum under standard conditions. Total RNA was extracted using the Tripure Isolation Reagent. The relative mRNA expressions of OCT4, ALDH1A1 and CD44 associated with stemness as well as TGF-β1, TβR1, ERα1 and MnSOD associated with aggressiveness in BCSCs and MCF-7 cells were determined using the quantitative real-time PCR (qRT-PCR).ResultsThe mRNA expressions of OCT4 (5.19-fold ± 0.338; P = 0.001), ALDH1A1 (3.67-fold ± 0.523; P = 0.006), CD44 (2.65-fold ± 0.307; P = 0.006), TGF-β1 (22.89-fold ± 6.840; P = 0.015), TβR1 (3.74-fold ± 1.446; P = 0.045) and MnSOD (4.6-fold ± 1.096; P = 0.014) were higher in BCSCs than in MCF-7 but were almost similar to MDA-MB-231 cells. In contrast, the ERα1 expression of BCSCs (0.97-fold ± 0.080; P = 0.392) was similar to MCF-7 cells, indicating that BSCSs are oestrogen-dependent breast cancer cells.ConclusionThe oestrogen-dependent BCSCs express stemness and aggressiveness genes at a higher level compared to oestrogen-dependent MCF-7 but are almost similar to oestrogen-independent MDA-MB-231 cells.

Xiaochaihu Decoction reduces hepatic steatosis and improves D-GalN/LPS-induced liver injury in hybrid grouper (Epinephelus lanceolatus♂ × Epinephelus fuscoguttatus♀)

Excessive lipid accumulation and chemical abuse can induce fatty liver diseases in fish, but the underlying mechanism and therapies are unknown. The present study aims to evaluate the effects of Xiaochaihu Decoction (XCHD) on the growth performance, lipid metabolism and antioxidant function of hybrid grouper in vitro and in vivo, and provide evidence as to whether it can be potentially used as a medicine for liver diseases in aquaculture. In vitro, steatosis model of hybrid grouper primary hepatocytes were incubated for 48 h in control or lipid emulsion (LE)-containing medium with or without 24 h post-treatment with XCHD. XCHD treatment reversed the LE-induced intracellular lipid accumulation, cell viability and hepatocytes morphological structure. In vivo, a total of 300 hybrid grouper with an average initial weight of 25.43 ± 0.18 g were fed diets containing five graded levels of XCHD at 150–1200 mg/kg diet for 8 weeks. After that, a challenge trial was conducted by injection of D-GalN/LPS to induce liver injury. As a result, dietary supplementation with 150–300 mg/kg XCHD diets can significant improve growth performance and feed utilization (P < 0.05). Dietary XCHD down-regulated the expression of lipogenic-related genes (G6PD, DGAT2 and ME1) and up-regulated lipolysis-related genes (ATGL, PPARα and LPL) expression in the liver of hybrid grouper. Livers challenged with D-GalN/LPS exhibited extensive areas of vacuolization with the disappearance of nuclei and the loss of hepatic architecture. These pathological alterations were ameliorated by XCHD treatment. XCHD significantly down-regulated the D-GalN/LPS induced apoptosis-related genes caspase-3, caspase-9 and p53 mRNA expression and up-regulated the antioxidant-related genes CAT and MnSOD mRNA expression in dose dependent manner, respectively. XCHD potently reduced hepatic lipid accumulation and enhanced antioxidant capability in hybrid grouper and may be a potential fish-feed additive to prevent fatty liver diseases onset and progression.

Wu Shan Shen Cha (Malus asiatica Nakai. Leaves)-Derived Flavonoids Alleviate Alcohol-Induced Gastric Injury in Mice via an Anti-Oxidative Mechanism.

Wu Shan Shen Cha is the leaf of Malus asiatica Nakai., a special type of tea that is consumed in the same way as green tea. To study the effect of Wu Shan Shen Cha-derived flavonoids (WSSCF) on lesions in the stomach, a 15% hydrochloric acid–95% ethanol (volume ratio 4:6) solution was used to induce gastric injury in mice. The degree of gastric injury was assessed using tissue specimens, and the effects of WSSCF on the serum levels of antioxidant enzymes were investigated. The results showed that WSSCF could alleviate the damage of the gastric mucosa and gastric wall caused by the hydrochloric acid–ethanol solution, decrease the tissue and serum levels of malondialdehyde (MDA) in mice with gastric injury, and increase the serum levels of superoxide dismutase (SOD) and glutathione (GSH). The results of quantitative polymerase chain reaction (qPCR) showed that WSSCF could increase the mRNA expression of Mn-SOD, Cu/Zn-SOD, catalase (CAT), endothelial nitric oxide synthase (eNOS), and neuronal nitric oxide synthase (nNOS) in tissue specimens from mice with gastric injury and decrease the expression of cyclooxygenase-2 (COX-2) and inducible nitric oxide synthase (iNOS). At the same time, the results of the high concentration of WSSCF (WSSCFH) group were closer to those of the drug (ranitidine) treatment group. Wu Shan Shen Cha-derived flavonoids had a good antioxidant effect, so as to play a preventive role in alcoholic gastric injury.

Efek Hipoxia Mimetic Cobalt Chloride (CoC12 )terhadap Ekspresi mRNA dan Aktivitas Spesifik Manganese Superoksida Dismutase (Mn SOD ) Ginjal Tikus

&lt;p&gt;MnSOD is a primary antioxidant that protects cells from oxidative stress due to H2O2 in&lt;br /&gt;mitochondrial membranes. This study aims to determine the effect of CoCl2 as hypoxic mimetic agent on mRNA expression of MnSOD and MnSOD-specific activity in rat kidney. Male Sprague dawley rat are induced with 30mg/Kg body weight CoCl2 intraperitoneally. After that, the experiment was divided into 3 groups: 2, 8, and 24 hours incubation after injection. All groups are compared with control group without injection. All of rat were terminated and the kidneys were removed. mRNA expression and specific activity of the kidney MnSOD are measured. After 2 hours, mRNA expression increased up to 42 times, after 8 and 24 hours return to normal. Specific activity of MnSOD in 2 hours after injection has not changed yet, and after 8 and 24 hours increased 1.5 times. This study shows that induction of Hipoxic mimetic CoCl2 enhances mRNA expression and MnSOD-specific activity.&lt;/p&gt;

DNA-Conjugated Amphiphilic Aggregation-Induced Emission Probe for Cancer Tissue Imaging and Prognosis Analysis.

Detection of an ultralow concentration of mRNA is important in the prognosis of gene-related diseases. In this study, a DNA-conjugated amphiphilic aggregation-induced emission probe (TPE-R-DNA) was synthesized for cancer tissue imaging and prognosis analysis based on an exonuclease III-aided target recycling technique. TPE-R-DNA comprise two components: a hydrophobic component that serves as the "turn-on" long wavelength fluorescence imaging agent (TPE-R-N3); and a hydrophilic single DNA strand (Alk-DNA) which acts as specific recognition part for target mRNA. In the absence of target mRNA, TPE-R-DNA had almost no fluorescence because of its high water solubility. Conversely, the TPE-R-DNA was digested by exonuclease III (Exo III) in the presence of MnSOD mRNA to release the hydrophobic fluorogens (TPE-R-AT). Subsequently, TPE-R-AT formed aggregates, and therefore, fluorescence signal was distinctly observed. For the first time, the structure of the hydrolysis product (TPE-R-AT), containing two bases A and T, was proved by the mass spectrum (MS) and high-performance liquid chromatography (HPLC). Moreover, the detection limit toward mRNA could be achieved in as low as 0.6 pM. Furthermore, the fluorescent signal can be used to confirm the MnSOD mRNA expression level in cancer tissue. The MnSOD mRNA expression in renal cancer was lower than in renal cancer adjacent tissue. In particular, the expression level was analyzed to predict prognosis of cancer patients. Our results demonstrate that a shorter survival time was evident among patients in lower MnSOD mRNA expression. Thereby, it indicates great potential for the development of an ultrasensitive biosensing platform for the application in disease prognosis.

The role of CuZn- and Mn-superoxide dismutases in earthworm Eisenia andrei kept in two distinct field-contaminated soils

Polychlorinated dibenzo-p-dioxins/dibenzofurans (PCDD/Fs), together with polycyclic aromatic hydrocarbons (PAHs), represent highly toxic and persistent organic environmental pollutants, especially due to their capability for bioaccumulation in fatty tissues. To observe the environmentally relevant effect of these compounds on earthworms, two soils naturally contaminated with PCDD/Fs and PAHs were used in our experiments. We focused on the role of CuZn- and Mn-superoxide dismutases. We assembled a full-length sequences of these molecules from Eisenia andrei earthworm and confirmed their activity. We demonstrated the significant reduction of CuZn-SOD on both mRNA and enzyme activity levels and increased levels of reactive oxygen species in earthworms kept in PCDD/F-polluted soil, which corresponds to the observed histopathologies of the earthworm intestinal wall and adjacent chloragogenous tissue. The results show an important role of CuZn-SOD in earthworm tissue damage caused by PCDD/Fs present in soil. We did not detect any significant changes in the mRNA expression or activity of Mn-SOD in these earthworms. In earthworms maintained in PAH-polluted soil the activity of both CuZn-SOD and Mn-SOD significantly increased. No histopathological changes were detected in these worms, however significant decrease of coelomocyte viability was observed. This reduced viability was most likely independent of oxidative stress.