scholarly journals Compound Dihuang Granule Protects against 6-OHDA Induced Toxicity in Parkinson’s Disease Rats by Suppressing the Phosphorylation of MAPK/ERK1/2

2021 ◽  
Author(s):  
Li Wang ◽  
Jian-ying Zhang ◽  
Long Chen ◽  
Lei Zhang ◽  
Zhu-qing He ◽  
...  
Keyword(s):  
Oxidative Stress ◽  
Substantia Nigra ◽  
Western Blotting ◽  
Neurotrophic Factors ◽  
Neurotrophic Factor ◽  
Cell Apoptosis ◽  
Colorimetric Method ◽  
Substantia Nigra Pars Compacta ◽  
Therapeutic Effects ◽  
Expression Levels

Abstract BackgroundParkinson’s disease (PD) is a multifactorial neurodegenerative disorder characterized by progressive loss of dopaminergic (DA) neurons in the substantia nigra pars compacta (SNpc) and the presence of Lewy bodies (LBs) consisting of misfolded α-synuclein protein in the substantia nigra pars compacta (SNpc). Compound Dihuang Granule (CDG), a famous traditional Chinese medicine (TCM) has been clinically used in PD therapy with curative effects. However, the specific functions and the mechanism of action remained unclear. This study explored the therapeutic effects and potential mechanisms of CDG in the PD rats induced by 6-OHDA toxicity.MethodsThe PD rat model was induced by unilaterally stereotactic injection of 6-OHDA into the SNpc of midbrain. The behavioral performances of rats were evaluated by rotation test, muscle strength assessment and balance beam walking test. The striatal contents of neurotransmitters were detected by HPLC.The numbers of dopaminergic (DA) neurons were determined with immunohistochemistry (IHC) staining and Western blotting assay. Indicators of oxidative stress were determined with colorimetric method. Apoptotic cells were detected by TUNEL assay. The expression levels of neurotrophic factors were examined with IHC staining and real-time quantitative PCR. The related protein expression levels were determined with Western blotting assay.ResultsCDG significantly attenuated the 6-OHDA induced abnormal rotational behaviors and alleviated the loss of DA neurons in the nigrostriatal axis of PD rats with a 6-week treatment. Consistently, the striatal contents of DA and its metabolites including DOPAC and HVA of PD rats were all significantly increased with CDG treatment. The 6-OHDA induced oxidative stress indicated with decreased superoxide dismutase (SOD), glutathione (GSH), glutathione peroxidase (GSH-Px) and increased malondialdehyde (MDA) was also suppressed by CDG treatment. Moreover, CDG treatment increased the expression levels of neurotrophic factors including nerve growth factor (NGF), brain-derived neurotrophic factor (BDNF) and glial-derived neurotrophic factor (GDNF) in the nigrostriatal axis of PD rats. Consistently, the 6-OHDA induced cell apoptosis was inhibited by the 6-week CDG treatment. Further, the phosphorylation of MAPK/ERK1/2 and CREB proteins in the striatum of PD rats was suppressed by CDG treatment and CDG showed synergistic effects with the MAPK/ERK1/2 phosphorylation inhibitor SL327.ConclusionCDG could ameliorate the 6-OHDA induced brain injuries and motor symptoms, and also inhibit the oxidative stress and cell apoptosis in the nigrostriatal axis, which was mainly mediated by enhancing the expression levels of the neurotrophic factors and suppressing the phosphorylation of MAPK/ERK1/2 pathway in the midbrain of rats.

scholarly journals Compound Dihuang Granule Attenuates the Da Neuron Loss and Motor Deficits in 6-OHDA  Induced Parkinson’s Disease Rats

2020 ◽  
Author(s):  
Li Wang ◽  
Dian-yong Bi ◽  
Zhu-qing He ◽  
Lei Zhang ◽  
Yu-fang Yang ◽  
...  
Keyword(s):  
Oxidative Stress ◽  
Substantia Nigra ◽  
Protein Expression ◽  
Neurotrophic Factors ◽  
Neurotrophic Factor ◽  
Cell Apoptosis ◽  
Substantia Nigra Pars Compacta ◽  
Motor Deficits ◽  
Apoptotic Cells ◽  
Expression Levels

Abstract Background: Parkinson’s disease (PD) is a multifactorial neurodegenerative disorder characterized by progressive loss of dopaminergic (DA) neurons in the substantia nigra pars compacta (SNpc) and the presence of Lewy bodies (LBs) consisting of misfolded α-synuclein protein in the substantia nigra pars compacta (SNpc). Compound Dihuang Granule (CDG), a famous traditional Chinese medicine (TCM) has been clinically used in PD therapy with curative effects. However, the specific functions and the mechanism of action remained unclear. This paper study assesses the preventive and therapeutic effect of CDG on motor deficits and DA neuron loss of PD induced by 6-OHDA and the underlying mechanisms.Methods: PD rat model was induced by unilaterally stereotactic injection of 6-OHDA into the SNpc of midbrain then the motor deficits were evaluated with apomorphine (APO) induced abnormal rotational behaviors. The striatal contents of neurotransmitters were detected by high performance liquid chromatography with electrochemical detection (HPLC-ECD). The number of DA neurons were determined with immunohistochemistry (IHC) staining. Protein expression levels were determined with Western blotting assay. Indicators of oxidative stress were determined with colorimetric method. Apoptotic cells were detected by Transferase-mediated deoxyuridine triphosphate-biotin nick end labeling (TUNEL) assay. The expression of neurotrophic factors was examined with IHC staining.Results: With a 6-week treatment, CDG significantly attenuated the 6-OHDA induced abnormal rotational behaviors and alleviated the loss of DA neurons in the nigrostriatal axis. Consistently, the striatal contents of DA and its metabolites including DOPAC and HVA of PD rats were all significantly increased with CDG treatment. The 6-OHDA induced oxidative stress indicated with decreased superoxide dismutase (SOD), glutathione (GSH), glutathione peroxidase (GSH-Px) and increased malondialdehyde (MDA)was also suppressed by CDG. Moreover, CDG treatment inhibited the 6-OHDA induced cell apoptosis indicated with decreased apoptotic cells in the SNpc and increased protein expression ratio of Bcl-2/Bax in the striatum. The expression levels of neurotrophic factors including nerve growth factor (NGF), brain-derived neurotrophic factor (BDNF) and glial-derived neurotrophic factor (GDNF)in the SNpc of PD rats were also increased by CDG.Conclusion: CDG could ameliorate the 6-OHDA induced brain injuries and motor symptoms mainly by inhibition of the oxidative stress and cell apoptosis in the nigrostriatal axis, and enhancing the expression of neurotrofic factors in the midbrain of rats.

scholarly journals Artemisinin attenuated oxidative stress and apoptosis by inhibiting autophagy in MPP+-treated SH-SY5Y cells

2021 ◽  
Vol 28 (1) ◽  
Author(s):  
Junqiang Yan ◽  
Hongxia Ma ◽  
Xiaoyi Lai ◽  
Jiannan Wu ◽  
Anran Liu ◽  
...  
Keyword(s):  
Oxidative Stress ◽  
Membrane Potential ◽  
Protein Expression ◽  
Mitochondrial Membrane Potential ◽  
Western Blotting ◽  
Cell Apoptosis ◽  
Mitochondrial Membrane ◽  
Caspase 3 ◽  
Neuroprotective Effect ◽  
Neuroprotective Effects

Abstract Background Parkinson’s disease (PD) is the second most common neurodegenerative disease after Alzheimer's disease. The oxidative stress is an important component of the pathogenesis of PD. Artemisinin (ART) has antioxidant and neuroprotective effects. The purpose of this study is to explore the neuroprotective effect of ART on 1-methyl-4-phenyliodine iodide (MPP +)-treated SH-SY5Y cells and underlying mechanism. Methods We used MPP+-treated SH-SY5Y cells to study the neuroprotective effect of ART. Cell viability was measured by MTT assay after incubating the cells with MPP+ and/or ART for 24 h. DCFH-DA was used to detect the level of intracellular reactive oxygen species (ROS), and WST-8 was used to detect the level of superoxide dismutase (SOD). The level of intracellular reduced glutathione (GSH) was detected with 5,5΄-dithiobis-(2-nitrobenzoic acid), and the level of malondialdehyde (MDA) was assessed based on the reaction of MDA and thiobarbituric acid. A mitochondrial membrane potential detection kit (JC-1) was used to detect changes in the mitochondrial membrane potential (MMP), and an Annexin V-FITC cell apoptosis kit was used to detect cell apoptosis. The expression levels of caspase-3, cleaved caspase-3 and the autophagy-related proteins LC3, beclin-1, and p62 were detected by Western blotting. In addition, to verify the change in autophagy, we used immunofluorescence to detect the expression of LC3 and p62. Results No significant cytotoxicity was observed at ART concentrations up to 40 μM. ART could significantly increase the viability of SH-SY5Y cells treated with MPP+ and reduce oxidative stress damage and apoptosis. In addition, the Western blotting and immunofluorescence results showed that MPP+ treatment could increase the protein expression of beclin1 and LC3II/LC3I and decrease the protein expression of p62, indicating that MPP+ treatment could induce autophagy. Simultaneous treatment with ART and MPP+ could decrease the protein expression of beclin1 and LC3II/LC3I and increase the protein expression of p62, indicating that ART could decrease the level of autophagy induced by MPP+. Conclusion Our results indicate that ART has a protective effect on MPP+-treated SH-SY5Y cells by the antioxidant, antiapoptotic activities and inhibition of autophagy. Our findings may provide new hope for the prevention and treatment of PD.

scholarly journals Oxymatrine Ameliorates Memory Impairment in Diabetic Rats by Regulating Oxidative Stress and Apoptosis: Involvement of NOX2/NOX4

2020 ◽  
Vol 2020 ◽  
pp. 1-15
Author(s):  
Yongpan Huang ◽  
Xinliang Li ◽  
Xi Zhang ◽  
Jiayu Tang
Keyword(s):  
Oxidative Stress ◽  
Reactive Oxygen Species ◽  
Brain Injury ◽  
Western Blotting ◽  
Caspase 3 ◽  
Memory Function ◽  
Diabetic Rats ◽  
Oxygen Species ◽  
Expression Levels

Oxymatrine (OMT) is the major quinolizidine alkaloid extracted from the root of Sophora flavescens Ait and has been shown to exhibit a diverse range of pharmacological properties. The aim of the present study was to investigate the role of OMT in diabetic brain injury in vivo and in vitro. Diabetic rats were induced by intraperitoneal injection of a single dose of 65 mg/kg streptozotocin (STZ) and fed a high-fat and high-cholesterol diet. Memory function was assessed using a Morris water maze test. A SH-SY5Y cell injury model was induced by incubation with glucose (30 mM/l) to simulate damage in vitro. The serum fasting blood glucose, insulin, serum S100B, malondialdehyde (MDA), and superoxide dismutase (SOD) levels were analyzed using commercial kits. Morphological changes were observed using Nissl staining and electron microscopy. Cell apoptosis was assessed using Hoechst staining and TUNEL staining. NADPH oxidase (NOX) and caspase-3 activities were determined. The effects of NOX2 and NOX4 knockdown were assessed using small interfering RNA. The expression levels of NOX1, NOX2, and NOX4 were detected using reverse transcription-quantitative PCR and western blotting, and the levels of caspase-3 were detected using western blotting. The diabetic rats exhibited significantly increased plasma glucose, insulin, reactive oxygen species (ROS), S-100B, and MDA levels and decreased SOD levels. Memory function was determined by assessing the percentage of time spent in the target quadrant, the number of times the platform was crossed, escape latency, and mean path length and was found to be significantly reduced in the diabetic rats. Hyperglycemia resulted in notable brain injury, including histological changes and apoptosis in the cortex and hippocampus. The expression levels of NOX2 and NOX4 were significantly upregulated at the protein and mRNA levels, and NOX1 expression was not altered in the diabetic rats. NOX and caspase-3 activities were increased, and caspase-3 expression was upregulated in the brain tissue of diabetic rats. OMT treatment dose-dependently reversed behavioral, biochemical, and molecular changes in the diabetic rats. In vitro, high glucose resulted in increases in reactive oxygen species (ROS), MDA levels, apoptosis, and the expressions of NOX2, NOX4, and caspase-3. siRNA-mediated knockdown of NOX2 and NOX4 decreased NOX2 and NOX4 expression levels, respectively, and reduced ROS levels and apoptosis. The results of the present study suggest that OMT alleviates diabetes-associated cognitive decline, oxidative stress, and apoptosis via NOX2 and NOX4 inhibition.

scholarly journals Protective Effects of Tianxiangdan Capsule Drug-Containing Plasma Against H2O2-Induced Oxidative Stress in Primary Cardiomyocytes

2021 ◽  
Author(s):  
Mei Tang ◽  
Lin Jiang ◽  
Gaerma Dugujia ◽  
Yuche Wu ◽  
Xiao Liu ◽  
...  
Keyword(s):  
Oxidative Stress ◽  
Myocardial Ischemia ◽  
Cell Apoptosis ◽  
Cell Damage ◽  
Network Pharmacology ◽  
Protective Effects ◽  
Active Components ◽  
Stress Injury ◽  
Expression Levels ◽  
Intracellular Ca

Abstract Background: Tianxiangdan capsule (TXD), developed in our hospital, has been clinically used in the treatment of coronary heart disease angina pectoris. This study aimed at evaluating the mechanisms of TXD against myocardial ischemia and to provide evidence for its subsequent clinical application. METHODS: Active components and mechanisms of action of TXD against myocardial ischemia were predicted and analyzed by network pharmacology and molecular docking. The oxidative damage model was established using H2O2, which caused myocardial cell damage. The MTT assay was used to evaluate cell viability, Hoechst33342 staining, while cleaved caspase-3 immunofluorescence staining was used to determine cell apoptosis. Fluorescent probe method detected ROS and intracellular Ca2+, while spectrophotometry was used to measure SOD, MDA, and NO levels in myocardial cells. Western blotting was used to detect the expression levels of ESR1, PI3K, AKT, and eNOS in cells. RESULTS: It was found that TXD plays a protective role in myocardial ischemia through the estrogen pathway, and its main active components were isoflavones. The TXD drug-containing plasma exhibited increased cell survival rates and suppressed MDA levels, elevated SOD and NO levels, and significantly suppressed ROS levels as well as intracellular Ca2+ levels. Moreover, the TXD drug-containing plasma pretreated cells had significantly suppressed PI3K and AKT expression levels, as well as elevated ESR1 and eNOS expression levels. Conclusion: TXD may exhibit estrogen-like effects, through the estrogen pathway enhances cardiomyocytes' antioxidant capacities, and improves oxidative stress injury as well as cell apoptosis.

scholarly journals IMP3 accelerates the progression of prostate cancer through inhibiting PTEN expression in a SMURF1-dependent way

2020 ◽  
Vol 39 (1) ◽  
Author(s):  
Xiang Zhang ◽  
Dawei Wang ◽  
Boke Liu ◽  
Xingwei Jin ◽  
Xianjin Wang ◽  
...  
Keyword(s):  
Prostate Cancer ◽  
Cell Viability ◽  
Signaling Pathway ◽  
Cell Lines ◽  
Western Blotting ◽  
Cell Apoptosis ◽  
Expression Level ◽  
Mtor Signaling Pathway ◽  
Expression Levels ◽  
Cancer Tissues

Abstract Background Insulin-like growth factor 2 (IGF2) messenger RNA binding protein 3 (IMP3) has been testified to be overexpressed in prostate cancer and strongly related to patients’ poor prognosis. However, the functions of IMP3 and the underlying mechanisms in prostate cancer still remain unknown. Therefore, the current study was carried out to reveal the role and molecular mechanism of IMP3 in prostate cancer progression. Methods The expression levels of IMP3 in prostate cancer tissues and cells were detected by immunohistochemistry (IHC), western blotting and RT-PCR. CCK-8, clone formation, flow cytometry and in vivo tumor formation assays were used to determine cell growth, clone formation apoptosis and tumorigenesis, respectively. The effect of IMP3 on the expression levels of the key proteins in PI3K/AKT/mTOR signaling pathway, including PIP2, PIP3, p-AKT, AKT, p-mTOR, mTOR, PTEN and BAD activation of was determined by western blotting. IP (Immunoprecipitation) assay was used to evaluate the effects of IMP3 and SMURF1 (SMAD specific E3 ubiquitin protein ligase 1) on the ubiquitination of PTEN protein. Results IMP3 expression level was significantly increased in prostate cancer tissues and cell lines (LNCap, PC3 and DU145) as compared with the paracancerous normal tissues and cells (RWPE-1), respectively. High expression of IMP3 apparently promoted cell viability, tumorigenesis and inhibited cell apoptosis in prostate cancer LNCap, DU145 and PC3 cell lines. In mechanism, IMP3 upregulation significantly increased the phosphorylation levels of AKT and mTOR, and elevated PIP3 expression level, while induced significant reductions in the expression levels of BAD, PTEN and PIP2. And, IMP3 overexpression increased SMURF1 expression, which facilitated PTEN ubiquitination. In addition, SMURF1 overexpression enhanced prostate cancer cell viability and inhibited cell apoptosis. Silence of SMURF1 rescued the enhancements in cell proliferation and tumorigenesis and the inhibition in cell apoptosis rates induced by IMP3 in prostate cancer DU145 and LNCap cells. Conclusion This study reveals that IMP3 is overdressed in prostate cancer, which accelerates the progression of prostate cancer through activating PI3K/AKT/mTOR signaling pathway via increasing SMURF1-mediated PTEN ubiquitination.

scholarly journals Cyclosporin A protects JEG-3 cells against oxidative stress-induced apoptosis by inhibiting the p53 and JNK/p38 signaling pathways

2020 ◽  
Vol 18 (1) ◽  
Author(s):  
Bin He ◽  
Qi Yue Li ◽  
Yuan Yuan Wu ◽  
Jing Ling Ruan ◽  
Xiao Ming Teng ◽  
...  
Keyword(s):  
Oxidative Stress ◽  
Cyclosporin A ◽  
Signaling Pathways ◽  
Western Blotting ◽  
Cell Apoptosis ◽  
Caspase 3 ◽  
B Cell Lymphoma ◽  
Trophoblast Invasion ◽  
Trophoblast Cells ◽  
Induced Apoptosis

Abstract Background Trophoblast cells are required for the establishment of pregnancy and fetal development. Apoptosis is an essential feature for trophoblast invasion. Uncontrolled trophoblast apoptosis is related to some complicate pregnancies. Oxidative stress (OS) is an important inducer of trophoblast apoptosis. Cyclosporin A (CsA) has been shown to promote the activity of trophoblast cells and reduce OS-induced oxidative injury. We investigated the role and mechanism of CsA in oxidative stress-induced trophoblast cell apoptosis. Methods JEG-3 cells were cocultured with H2O2 and CsA. Cell viability and morphology were measured by MTT assay and DAPI staining. Cell apoptosis was tested with annexin V/PI staining. The expression of Bcl-2-associated X protein (Bax), B-cell lymphoma/leukemia-2 (Bcl-2), cleaved poly (ADP-ribose) polymerase (PARP) and pro-caspase-3 was assayed by western blotting. The protein expression and phosphorylation of p53 and mitogen-activated protein kinase (MAPK) kinases (JNK, ERK1/2 and p38) were examined by western blotting. Results CsA increased the viability, alleviated morphological injury and reduced cell apoptosis of the H2O2-treated JEG-3 cells. CsA also attenuated the activation of p53, decreased the expression of Bax and cleavage of PARP, and increased the expression of Bcl-2 and pro-caspase-3 in the JEG-3 treated with H2O2. Furthermore, CsA reduced the activation of JNK and P38 but had no significant effect on the activation of extracellular signal-regulated kinase 1/2 (ERK1/2) in the H2O2-treated JEG-3 cells. Promoting the activation of JNK and p38 impaired the protective effect of CsA on OS-induced trophoblast apoptosis. Conclusions These results suggested that CsA protected trophoblast cells from OS-induced apoptosis via the inhibition of the p53 and JNK/p38 signaling pathways.

Downregulation of General Control Nonderepressible 2/Eukaryotic Initiation Factor 2α Attenuates Inflammation, Oxidative Stress and Cell Apoptosis in Retinal Cells Induced by High Glucose

2021 ◽  
Vol 11 (8) ◽  
pp. 1497-1505
Author(s):  
Shuyu Zhao ◽  
Yuqian Yin ◽  
Hong Qin
Keyword(s):  
Oxidative Stress ◽  
Endothelial Cell ◽  
Western Blot ◽  
High Glucose ◽  
Cell Apoptosis ◽  
Colorimetric Method ◽  
Underlying Mechanism ◽  
Initiation Factor ◽  
Western Blot Assay ◽  
And Oxidative Stress

Background: Diabetic retinopathy (DR), the frequent complication of diabetes mellitus, has been the main factor of clinical blindness. It is of great clinical significance to seek a novel therapeutic target of DR. The present study aims to investigate the important role of GCN2/eIF2α in DR and the underlying mechanism. Materials and Methods: The expression levels of GCN2 and p-eIF2α were measured by western blot assay and q-PCR. The inflammation levels were assessed by ELISA assay and oxidative stress was measured by colorimetric method. Then, the key proteins related to the function of endothelial cell were measured by western blot assay. Cell apoptotic rate was detected by flow cytometry and proteins related to cell apoptosis were detected by western blot assay. Results: High glucose activated GCN2/eIF2α signaling pathway in HRCECs. Downregulation of GCN2 attenuated HG-induced cell apoptosis, inflammatory and oxidative stress in HRCECs. Meanwhile, downregulation of GCN2 ameliorated HG-induced endothelial cell dysfunction. Inhibition of GCN2 inhibited p-eIF2α, ATF4, CHOP and activated UCP2. Conclusion: The results in this study proved that knockdown of GCN2 could significantly mitigate HG-induced injury, suggesting GCN2/eIF2α as a potential target for DR therapy.

scholarly journals Copper and Copper Proteins in Parkinson’s Disease

2014 ◽  
Vol 2014 ◽  
pp. 1-15 ◽  
Author(s):  
Sergio Montes ◽  
Susana Rivera-Mancia ◽  
Araceli Diaz-Ruiz ◽  
Luis Tristan-Lopez ◽  
Camilo Rios
Keyword(s):  
Oxidative Stress ◽  
Parkinson’S Disease ◽  
Parkinson's Disease ◽  
Substantia Nigra ◽  
Alpha Synuclein ◽  
Substantia Nigra Pars Compacta ◽  
Copper Binding ◽  
Copper Proteins ◽  
Limited Information ◽  
Copper Chelation

Copper is a transition metal that has been linked to pathological and beneficial effects in neurodegenerative diseases. In Parkinson’s disease, free copper is related to increased oxidative stress, alpha-synuclein oligomerization, and Lewy body formation. Decreased copper along with increased iron has been found insubstantia nigraand caudate nucleus of Parkinson’s disease patients. Copper influences iron content in the brain through ferroxidase ceruloplasmin activity; therefore decreased protein-bound copper in brain may enhance iron accumulation and the associated oxidative stress. The function of other copper-binding proteins such as Cu/Zn-SOD and metallothioneins is also beneficial to prevent neurodegeneration. Copper may regulate neurotransmission since it is released after neuronal stimulus and the metal is able to modulate the function of NMDA and GABA A receptors. Some of the proteins involved in copper transport are the transporters CTR1, ATP7A, and ATP7B and the chaperone ATOX1. There is limited information about the role of those biomolecules in the pathophysiology of Parkinson’s disease; for instance, it is known that CTR1 is decreased insubstantia nigra pars compactain Parkinson’s disease and that a mutation in ATP7B could be associated with Parkinson’s disease. Regarding copper-related therapies, copper supplementation can represent a plausible alternative, while copper chelation may even aggravate the pathology.

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