1.REEP1 Preserves Motor Function in SOD1G93A Mice by Improving Mitochondrial Function via Interaction with NDUFA4.
Siyue QIN ; Pan YOU ; Hui YU ; Bo SU
Neuroscience Bulletin 2023;39(6):929-946
A decline in the activities of oxidative phosphorylation (OXPHOS) complexes has been consistently reported in amyotrophic lateral sclerosis (ALS) patients and animal models of ALS, although the underlying molecular mechanisms are still elusive. Here, we report that receptor expression enhancing protein 1 (REEP1) acts as an important regulator of complex IV assembly, which is pivotal to preserving motor neurons in SOD1G93A mice. We found the expression of REEP1 was greatly reduced in transgenic SOD1G93A mice with ALS. Moreover, forced expression of REEP1 in the spinal cord extended the lifespan, decelerated symptom progression, and improved the motor performance of SOD1G93A mice. The neuromuscular synaptic loss, gliosis, and even motor neuron loss in SOD1G93A mice were alleviated by increased REEP1 through augmentation of mitochondrial function. Mechanistically, REEP1 associates with NDUFA4, and plays an important role in preserving the integrity of mitochondrial complex IV. Our findings offer insights into the pathogenic mechanism of REEP1 deficiency in neurodegenerative diseases and suggest a new therapeutic target for ALS.
Mice
;
Animals
;
Amyotrophic Lateral Sclerosis/metabolism*
;
Superoxide Dismutase-1/metabolism*
;
Superoxide Dismutase/metabolism*
;
Mice, Transgenic
;
Spinal Cord/pathology*
;
Mitochondria/physiology*
;
Disease Models, Animal
2.Shielding of the geomagnetic field reduces hydrogen peroxide production in human neuroblastoma cell and inhibits the activity of CuZn superoxide dismutase.
Hai-Tao ZHANG ; Zi-Jian ZHANG ; Wei-Chuan MO ; Ping-Dong HU ; Hai-Min DING ; Ying LIU ; Qian HUA ; Rong-Qiao HE
Protein & Cell 2017;8(7):527-537
Accumulative evidence has shown the adverse effects of a geomagnetic field shielded condition, so called a hypomagnetic field (HMF), on the metabolic processes and oxidative stress in animals and cells. However, the underlying mechanism remains unclear. In this study, we evaluate the role of HMF on the regulation of cellular reactive oxygen species (ROS) in human neuroblastoma SH-SY5Y cells. We found that HMF exposure led to ROS decrease, and that restoring the decrease by additional HO rescued the HMF-enhanced cell proliferation. The measurements on ROS related indexes, including total anti-oxidant capacity, HO and superoxide anion levels, and superoxide dismutase (SOD) activity and expression, indicated that the HMF reduced HO production and inhibited the activity of CuZn-SOD. Moreover, the HMF accelerated the denaturation of CuZn-SOD as well as enhanced aggregation of CuZn-SOD protein, in vitro. Our findings indicate that CuZn-SOD is able to response to the HMF stress and suggest it a mediator of the HMF effect.
Cell Line, Tumor
;
Humans
;
Hydrogen Peroxide
;
metabolism
;
Magnetic Fields
;
Neoplasm Proteins
;
metabolism
;
Neuroblastoma
;
metabolism
;
Stress, Physiological
;
Superoxide Dismutase-1
;
metabolism
3.Neuroprotection via maintenance or increase of antioxidants and neurotrophic factors in ischemic gerbil hippocampus treated with tanshinone I.
Joon Ha PARK ; Ok Kyu PARK ; Bingchun YAN ; Ji Hyeon AHN ; In Hye KIM ; Jae-Chul LEE ; Seung-Hae KWON ; Ki-Yeon YOO ; Choong Hyun LEE ; In Koo HWANG ; Jung Hoon CHOI ; Moo-Ho WON ; Jong-Dai KIM
Chinese Medical Journal 2014;127(19):3396-3405
BACKGROUNDDanshen (Radix Salvia miltiorrhizae) has been used as a traditional medicine in Asia for treatment of various microcirculatory disturbance related diseases. Tanshinones are mainly hydrophobic active components, which have been isolated from Danshen and show various biological functions. In this study, we observed the neuroprotective effect of tanshinone I (TsI) against ischemic damage in the gerbil hippocampal CA1 region (CA1) after transient cerebral ischemia and examined its neuroprotective mechanism.
METHODSThe gerbils were divided into vehicle-treated-sham-group, vehicle-treated-ischemia-group, TsI-treated-sham-group, and TsI-treated-ischemia-group. TsI was administrated intraperitoneally three times (once a day for three days) before ischemia-reperfusion. The neuroprotective effect of TsI was examined using H&E staining, neuronal nuclei (NeuN) immunohistochemistry and Fluoro-Jade B staining. To investigate the neuroprotective mechanism of TsI after ischemia-reperfusion, immunohistochemical (IHC) and Western blotting analyses for Cu, Zn-superoxide dismutase (SOD1), Mn-superoxide dismutase (SOD2), brain-derived neurotrophic factor (BDNF) and insulin-like growth factor-I (IGF-I) were performed.
RESULTSTreatment with TsI protected pyramidal neurons from ischemia-induced neuronal death in the CA1 after ischemia-reperfusion. In addition, treatment with TsI maintained the levels of SOD1 and SOD2 as determined by IHC and Western blotting in the CA1 after ischemia-reperfusion compared with the vehicle-ischemia-group. In addition, treatment with TsI increased the levels of BDNF and IGF-I determined by IHC and Western blotting in the TsI-treated-sham-group compared with the vehicle-treated-sham-group, and their levels were maintained in the stratum pyramidale of the ischemic CA1 in the TsI-treated-ischemia-group.
CONCLUSIONTreatment with TsI protects pyramidal neurons of the CA1 from ischemic damage induced by transient cerebral ischemia via the maintenance of antioxidants and the increase of neurotrophic factors.
Animals ; Antioxidants ; metabolism ; Blotting, Western ; Brain Ischemia ; drug therapy ; metabolism ; Brain-Derived Neurotrophic Factor ; metabolism ; Diterpenes, Abietane ; therapeutic use ; Gerbillinae ; Hippocampus ; metabolism ; Immunohistochemistry ; Insulin-Like Growth Factor I ; metabolism ; Male ; Nerve Growth Factors ; metabolism ; Superoxide Dismutase ; metabolism ; Superoxide Dismutase-1
4.Increased orexin expression promotes sleep/wake disturbances in the SOD1-G93A mouse model of amyotrophic lateral sclerosis.
Rong LIU ; Zhao-Fu SHENG ; Bing CAI ; Yong-He ZHANG ; Dong-Sheng FAN
Chinese Medical Journal 2015;128(2):239-244
BACKGROUNDSleep/wake disturbances in patients with amyotrophic lateral sclerosis (ALS) are well-documented, however, no animal or mechanistic studies on these disturbances exist. Orexin is a crucial neurotransmitter in promoting wakefulness in sleep/wake regulation, and may play an important role in sleep disturbances in ALS. In this study, we used SOD1-G93A transgenic mice as an ALS mouse model to investigate the sleep/wake disturbances and their possible mechanisms in ALS.
METHODSElectroencephalogram/electromyogram recordings were performed in SOD1-G93A transgenic mice and their littermate control mice at the ages of 90 and 120 days, and the samples obtained from these groups were subjected to quantitative reverse transcriptase-polymerase chain reaction, western blotting, and enzyme-linked immunosorbent assay.
RESULTSFor the first time in SOD1-G93A transgenic mice, we observed significantly increased wakefulness, reduced sleep time, and up-regulated orexins (prepro-orexin, orexin A and B) at both 90 and 120 days. Correlation analysis confirmed moderate to high correlations between sleep/wake time (total sleep time, wakefulness time, rapid eye movement [REM] sleep time, non-REM sleep time, and deep sleep time) and increase in orexins (prepro-orexin, orexin A and B).
CONCLUSIONSleep/wake disturbances occur before disease onset in this ALS mouse model. Increased orexins may promote wakefulness and result in these disturbances before and after disease onset, thus making them potential therapeutic targets for amelioration of sleep disturbances in ALS. Further studies are required to elucidate the underlying mechanisms in the future.
Amyotrophic Lateral Sclerosis ; genetics ; metabolism ; Animals ; Female ; Intracellular Signaling Peptides and Proteins ; genetics ; metabolism ; Male ; Mice ; Mice, Transgenic ; Neuropeptides ; genetics ; metabolism ; Orexins ; Reverse Transcriptase Polymerase Chain Reaction ; Sleep ; physiology ; Superoxide Dismutase ; genetics ; metabolism ; Superoxide Dismutase-1 ; Wakefulness ; physiology
5.Pro-oxidation status of rotator cuff tissue and expression of Beclin-1 and mTOR in patients with rotator cuff injury.
Zhi-Jun LIU ; He-Wei WEI ; Wei-Peng ZHENG ; Lei WAN ; Zhi-Hao LIAO ; Sheng CHEN
China Journal of Orthopaedics and Traumatology 2023;36(12):1136-1141
OBJECTIVE:
To explore pro-oxidative state of rotator cuff tissue and expression levels of Beclin-1 and mam-malian target of rapamycin(mTOR) in patients with acute and chronic rotator cuff injury, and then analyzed relationship between rotator cuff injury and oxidative stress and autophagy.
METHODS:
Forty patients with rotator cuff injury were seleceted from July 2019 to December 2020, and divided into male chronic injury group, male acute injury group, female chronic injury group, and female acute injury group, 10 patients in each group. All patients were performed rotator cuff repair under arthroscopy. The sample of tendon at the rotator cuff injury site of the patient was taken during operation, and total reactive oxygen species (ROS) and superoxide dismutase(SOD) were detected by detection kit;expression of Beclin-1 and mTOR mRNA were detected by reverse transcription-polymerase chain reaction (RT-PCR), and Western-blot was applied to detect protein expression of Beclin-1 and p-mTOR/mTOR.
RESULTS:
There were no significant difference in expression of ROS, SOD, Beclin-1mRNA and mTOR mRNA between male and female chronic injury groups, and between male and female acute injury groups (P>0.05); ROS, SOD and Beclin-1mRNA in male chronic injury group were higher than those in male chronic injury group, while mTOR mRNAand protein decreased (P<0.05);ROS, SOD and Beclin-1 mRNA in female chronic injury group were up-regulated compared with female acute injury group, while mTOR mRNA was down-regulated (P<0.05).
CONCLUSION
Chronic rotator cuff injury is more likely to stimulate the pro-oxidation state of rotator cuff tissue than acute rotator cuff injury, which could up-regulating expression of autophagy factor Beclin-1 and down-regulating expression of mTOR. Therefore, patients with chronic rotator cuff injury may have higher levels of oxidative stress and autophagy.
Female
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Humans
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Male
;
Beclin-1/metabolism*
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Reactive Oxygen Species/metabolism*
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RNA, Messenger/metabolism*
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Rotator Cuff/surgery*
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Rotator Cuff Injuries/surgery*
;
Superoxide Dismutase/metabolism*
;
TOR Serine-Threonine Kinases/metabolism*
6.Protective Effects of Anthocyanins Extracted from Vaccinium Uliginosum on 661W Cells Against Microwave-Induced Retinal Damage.
Lan YIN ; Si-Jun FAN ; Mao-Nian ZHANG
Chinese journal of integrative medicine 2022;28(7):620-626
OBJECTIVE:
To study the protective effect of anthocyanins extracted from Vaccinium Uliginosum (VU) on retinal 661W cells against microwave radiation induced retinal injury.
METHODS:
661W cells were divided into 6 groups, including control, model [661W cells radiated by microwave (30 mW/cm2, 1 h)] and VU groups [661W cells pretreated with anthocyanins extracted from VU (25, 50, 100 and 200 µg/mL, respectively) for 48 h, and radiated by microwave 30 mW/cm2, 1 h]. After treatment with different interventions, the cell apoptosis index (AI) was determined using Heochst staining; contents of malonaldehyde (MDA), glutataione (GSH), and activity of superoxide dismutase (SOD) were measured. mRNA expressions of nuclear factor erythroid 2-related factor 2 (Nrf2) and heme oxygenase 1(HO-1) were detected by real time quantitative polymerase chain reaction, and the expression of HO-1 protein was examined by Western blot analysis. Nucleus and cytoplasm were separated and Nrf2 protein expression was further verified by Western blot analysis.
RESULTS:
There was significant difference in AI among the groups (F=322.83, P<;0.05). Compared with the control group, AI was significantly higher in the model group and was lower in 4 VU-pretreated groups (P<;0.05). Linear regression analysis showed the decline of AI was in a dose-dependent manner with VU treatment (r=0.8419, P<;0.05). The MDA and GSH contents of 661W cells in VU-treated groups were significantly lower than the model group (P<;0.05). Compared with the model group, the SOD activity in the VU-treated groups (50, 100 and 200 µg/mL) was significantly higher (all P<;0.05). The Nrf2 and HO-1 mRNA expressions were slightly increased after irradiation, and obviously increased in 100 µg/mL VU-treated group. After irradiation, the relative expressions of HO-1 and Nrf2 proteins in nucleus were slightly increased (P<;0.05), and the changes in cytoplasm were not obvious, whereas it was significantly increased in both nucleus and cytoplasm in the VU treatment groups.
CONCLUSIONS
Anthocyanins extracted from VU could reduce apoptosis, stabilize cell membrane, and alleviate oxidant injury of mouse retinal photoreceptor 661W cells. The mechanism might be through activating Nrf2/HO-1 signal pathway and inducing HO-1 transcription and translation.
Animals
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Anthocyanins/therapeutic use*
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Blueberry Plants/metabolism*
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Heme Oxygenase-1/metabolism*
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Mice
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Microwaves
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NF-E2-Related Factor 2/metabolism*
;
Oxidative Stress
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RNA, Messenger/metabolism*
;
Superoxide Dismutase/metabolism*
7.Expression, purification, and characterization of cell-permeable fusion antioxidant enzyme sensitive to matrix metalloproteinases-2/9.
Huocong HE ; Lixiang LIN ; Lingling LI ; Lunqiao WU ; Haiying LIN ; Jianru PAN
Chinese Journal of Biotechnology 2022;38(9):3515-3527
Antioxidant enzymes fused with cell-penetrating peptides could enter cells and protect cells from irradiation damage. However, the unselective transmembrane ability of cell-penetrating peptide may also bring antioxidant enzymes into tumor cells, thus protecting tumor cells and consequently reducing the efficacy of radiotherapy. There are active matrix metalloproteinase (MMP)-2 or MMP-9 in most tumor cellular microenvironments. Therefore, a fusion protein containing an MMP-2/9 cleavable substrate peptide X, a cell-penetrating peptide R9, a glutathione S-transferase (GST), and a human Cu, Zn superoxide dismutase (SOD1), was designed and named GST-SOD1-X-R9. In the tumor microenvironment, GST-SOD1-X-R9 would lose its cell-penetrating peptide and could not enter tumor cells due to the cleavage of substrate X by active MMP-2/9, thereby achieving selected entering normal cells. The complete nucleotide sequence of SOD1-X-R9 was synthesized and inserted into the prokaryotic expression vector pGEX-4T-1. The pGEX4T-1-SOD1-X-R9 recombinant plasmid was obtained, and soluble expression of the fusion protein was achieved. GST-SOD1-X-R9 was purified by ammonium sulfate precipitation and GST affinity chromatography. The molecular weight of the fusion protein was approximately 47 kDa, consistent with the theoretical value. The SOD and GST activities were 2 954 U/mg and 328 U/mg, respectively. Stability test suggested that almost no change in either SOD activity or GST activity of GST-SOD1-X-R9 was observed under physiological conditions. The fusion protein could be partially digested by collagenase Ⅳ in solution. Subsequently, the effect of MMP-2/9 activity on transmembrane ability of the fusion protein was tested using 2D and 3D cultured HepG2 cells. Little extracellular MMP-2 activity of HepG2 cells was observed under 2D culture condition. While under the 3D culture model, the size and the MMP-2 activity of the HepG2 tumor spheroid increased daily. GST-SOD1-R9 proteins showed the same transmembrane efficiency in 2D cultured HepG2 cells, but the transmembrane efficiency of GST-SOD1-X-R9 in 3D cultured HepG2 spheres was reduced remarkably. This study provided a basis for further investigating the selectively protective effect of GST-SOD1-X-R9 against oxidative damage in normal cells.
Ammonium Sulfate
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Antioxidants
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Cell-Penetrating Peptides/pharmacology*
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Endopeptidases
;
Glutathione Transferase/metabolism*
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Humans
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Matrix Metalloproteinase 2/genetics*
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Matrix Metalloproteinase 9/genetics*
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Recombinant Fusion Proteins
;
Recombinant Proteins
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Superoxide Dismutase/metabolism*
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Superoxide Dismutase-1
8.Ulinastatin attenuates lung injury in rats with hemorrhagic shock.
Chun-shui LIN ; Peng LIU ; Ya-juan ZHAO ; Miao-ning GU ; Feng-yong XIE
Journal of Southern Medical University 2009;29(5):876-879
OBJECTIVETo investigate the effects of ulinastatin on lung injury in hemorrhagic shock rats.
METHODSTwenty-four normal SD rats were randomly divided into 3 groups (n=8), namely the control group, hemorrhagic shock group (group H) and ulinastatin group (group U). In group H and group U, blood was drawn from the femoral artery over a period of 10 min until a mean arterial pressure of 40 mmHg was obtained. Controlled hypotension was then maintained at 40-/+5 mmHg for 60 min by blood drawing or infusion when necessary. All the blood drawn and an equivalent volume of Ringer lactate solution were subsequently infused for resuscitation. Four hours after the resuscitation, the activity of superoxidedismutase (SOD), content of malondialdehyde (MDA), expression of heme oxygenase-1 (HO-1), wet to dry weight ratio (W/D), and pathologic changes of the lung tissues were measured or observed.
RESULTSCompared with those in the control group, the content of MDA, expression of HO-1 and W/D increased significantly in both group H and group U (P<0.05); these indexes in group U were significantly lower than those in group H (P<0.05). The activity of SOD in group U was significantly lower than that in the control group (P<0.05) but higher than that in group H (P<0.05). Optical microscopy demonstrated milder inflammatory cell infiltration and interstitial edema in the lung tissues in group U than in group H.
CONCLUSIONUlinastatin can lower the content of MDA, W/D and the expression of HO-1, increase the activity of SOD, and reduce histological lung injury in rats with hemorrhagic shock.
Animals ; Glycoproteins ; pharmacology ; Heme Oxygenase-1 ; metabolism ; Lung Injury ; etiology ; prevention & control ; Male ; Malondialdehyde ; metabolism ; Random Allocation ; Rats ; Rats, Sprague-Dawley ; Shock, Hemorrhagic ; complications ; metabolism ; Superoxide Dismutase ; metabolism
9.Polydatin improves intestinal barrier injury after traumatic brain injury in rats by reducing oxidative stress and inflammatory response via activating SIRT1-mediated deacetylation of SOD2 and HMGB1.
Na QIN ; Lin HUANG ; Rui DONG ; Fen LI ; Xu Heng TANG ; Zhen Hua ZENG ; Xing Min WANG ; Hong YANG
Journal of Southern Medical University 2022;42(1):93-100
OBJECTIVE:
To investigate the protective effect against intestinal mucosal injury in rats following traumatic brain injury (TBI) and explore the underlying mechanism.
METHODS:
SD rat models of TBI were established by fluid percussion injury (FPI), and the specimens were collected at 12, 24, 48, and 72 h after TBI. Another 15 rats were randomly divided into shamoperated group (n=5), TBI with saline treatment (TBI+NS) group (n=5), and TBI with PD treatment (TBI+PD) group (treated with 30 mg/kg PD after TBI; n=5). Body weight gain and fecal water content of the rats were recorded, and after the treatments, the histopathology of the jejunum was observed, and the levels of D-lactic acid (D-LAC), diamine oxidase (DAO), ZO-1, claudin-5, and reactive oxygen species (ROS) were detected. Lipid peroxide (LPO) and superoxide dismutase (SOD) 2 content, jejunal pro-inflammatory factors (IL-6, IL-1β, and TNF- α), Sirt1 activity, SOD2 and HMGB1 acetylation level were also determined after the treatments.
RESULTS:
The rats showed significantly decreased body weight and fecal water content and progressively increased serum levels of D-LAC and DAO after TBI (P < 0.05) with obvious jejunal injury, significantly decreased expression levels of ZO-1 and claudin-5, lowered SOD2 and Sirt1 activity (P < 0.05), increased expression levels of LPO, ROS, and pro-inflammatory cytokines, and enhanced SOD2 and HMGB1 acetylation levels (P < 0.05). Compared with TBI+NS group, the rats in TBI+PD group showed obvious body weight regain, increased fecal water content, reduced jejunal pathologies, decreased D-LAC and DAO levels (P < 0.05), increased ZO-1, claudin-5, SOD2 expression levels and Sirt1 activity, and significantly decreased ROS, LPO, pro-inflammatory cytokines, and acetylation levels of SOD2 and HMGB1 (P < 0.05).
CONCLUSION
PD alleviates oxidative stress and inflammatory response by activating Sirt1-mediated deacetylation of SOD2 and HMGB1 to improve intestinal mucosal injury in TBI rats.
Animals
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Brain Injuries, Traumatic
;
Glucosides/pharmacology*
;
HMGB1 Protein/metabolism*
;
Oxidative Stress
;
Rats
;
Rats, Sprague-Dawley
;
Sirtuin 1/metabolism*
;
Stilbenes/pharmacology*
;
Superoxide Dismutase/metabolism*
10.Danshensu delays the senescence of rat aortic endothelial cells via activation of SIRT1-SOD pathway.
Shuo WANG ; ; Di WU ; Lei LIU ; Jie CUI ; Wei-Li QIAO ; Hong SUN ; Chang-Dong YAN
Acta Physiologica Sinica 2014;66(5):575-582
The present study was aimed to investigate the effect of pretreatment with Danshensu (DSS) on rat aortic endothelial cells (RAECs) senescence and the underlying mechanisms. Cultured RAECs at fourth and twelfth passages were taken as young and old groups, respectively. DSS and DSS+nicotinamide (DSS+N) groups were incubated with DSS and DSS in combination with nicotinamide, an inhibitor of silent information regulator 1 (SIRT1), from the fourth to twelfth passage, respectively. The cell status of senescence was determined by the senescence-associated β-galactosidase (SA β-gal) staining, and 4,6-diamino-2-phenyl indole (DAPI) fluorescent dye was used to detect senescence associated heterochromatin foci (SAHF) formation; Thiobarbituric acid (TBA) and colorimetric methods were used to evaluate malondialdehyde (MDA) and H₂O₂contents; Western blot was employed to analysis the expressions of xanthine oxidase (XOD), SIRT1 and superoxide dismutase 2 (SOD₂) in the RAECs. The results showed that, in comparison with young group, the old group exhibited higher SA β-gal positive and SAHF formation rates, as well as higher MDA and H₂O₂levels (P < 0.05 or P < 0.01), whereas DSS pretreatment reduced SA β-gal positive and SAHF formation rates, decreased MDA and H2O2 contents (P < 0.05 or P < 0.01). The protection of DSS was reversed by nicotinamide. Compared with the young group, the old group showed higher expression levels of XOD, but lower SIRT1 and SOD₂expression levels (P < 0.05 or P < 0.01). With the pretreatment of DSS, the expression of XOD was declined, and the expression levels of SIRT1 and SOD₂were elevated, while nicotinamide reversed the effects of DSS. These results suggest that DSS delays senescence of RAECs via up-regulation of SIRT1.
Animals
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Aorta
;
cytology
;
Cells, Cultured
;
Cellular Senescence
;
drug effects
;
Endothelial Cells
;
cytology
;
Hydrogen Peroxide
;
metabolism
;
Lactates
;
pharmacology
;
Niacinamide
;
pharmacology
;
Rats
;
Sirtuin 1
;
metabolism
;
Superoxide Dismutase
;
metabolism
;
Up-Regulation