Hydroxychloroquine Blocks Autophagy and Promotes Apoptosis of the Prostate after Castration in Rats

2020 ◽  
Vol 104 (11-12) ◽  
pp. 968-974
Author(s):  
Sheng-Tao Ling ◽  
Chun-Lei Deng ◽  
Li Huang ◽  
Qi-Sheng Yao ◽  
Cui Liu ◽  
...  
Keyword(s):  
Caspase 3 ◽  
Morphological Changes ◽  
Control Group ◽  
Sham Operation ◽  
Related Gene ◽  
Rt Pcr ◽  
Bax Protein ◽  
Sd Rats ◽  
Immunohistochemical Analyses ◽  
Scanning Electron

Autophagy is an important pro-survival mechanism and closely related to apoptosis. The aim of this study was to investigate whether hydroxychloroquine (HCQ) blocks autophagy and promotes apoptosis of the prostate after castration. <b><i>Methods:</i></b> Thirty-six male SD rats were randomly divided into 3 groups (<i>n</i> = 12): control group (sham operation), castration group, and HCQ group (castrated and treated with HCQ). On day 7, all mice were executed and prostates were isolated. The morphological changes of prostates were observed by light microscope, and the ultrastructure changes were observed under scanning electron microscope (SEM). The protein expression of Beclin-l, P62, caspase-3, Bcl-2, and Bax was assessed by immunohistochemical analyses. The mRNA expression of microtubule-associated protein light chain 3 (LC3) and autophagy-related gene 5 (Atg5) was detected by RT-PCR. <b><i>Results:</i></b> Prostates of castration group shrank remarkably and prostates of HCQ group shrank more remarkably than castration group. Cytolysosomes were visible in the prostates of the castration group under SEM. Immunohistochemistry showed that the protein of Beclin-1 increased in the castration group compared to the control group, while decreased in the HCQ group compared to the castration group. While P62 protein moderately dyed in the control group and weakly dyed in the castration group, it strongly dyed in the HCQ group. Caspase-3 and Bax protein were weakly dyed in the control group but moderately dyed in the castration group and strongly dyed in the HCQ group. The expressions of apoptosis suppressor Bcl-2 were reduced in the castration group and further reduced in the HCQ group compared to the castration group. RT-PCR revealed that the mRNA of LC3 and Atg5 in the castration group increased compared to the control group, while decreased after treated with HCQ. <b><i>Conclusion:</i></b> Autophagy increased after castrated in prostates, while decreased after treated with HCQ; all these indicated that HCQ blocked autophagy and then promoted prostate apoptosis of castrated mice.

scholarly journals Preconditioning with L-alanyl-glutamine reduces hepatic ischemia-reperfusion injury in rats

2011 ◽  
Vol 26 (suppl 1) ◽  
pp. 8-13 ◽  
Author(s):  
Raimundo José Cunha Araújo Júnior ◽  
Raimundo Gerônimo da Silva Júnior ◽  
Marcelo Pinho Pessoa de Vasconcelos ◽  
Sérgio Botelho Guimarães ◽  
Paulo Roberto Leitão de Vasconcelos ◽  
...  
Keyword(s):  
Caspase 3 ◽  
Ischemia Reperfusion Injury ◽  
Ischemia Reperfusion ◽  
Control Group ◽  
Average Weight ◽  
Sham Operation ◽  
Hepatic Ischemia ◽  
Portal Triad ◽  
Hepatic Ischemia Reperfusion ◽  
Control Sham

PURPOSE: To evaluate the effects of pre-conditioning with L-alanyl- glutamine (L-Ala-Gln) in rats subjected to total hepatic ischemia. METHODS: Thirty Wistar rats, average weight 300g, were randomly assigned to 3 groups (n=10): G-1 - Saline, G-2- L-Ala-Gln, G-3-control (Sham). G-1 and G-3 groups were treated with saline 2.0 ml or L-Ala-Gln (0.75mg/Kg) intraperitoneally (ip) respectively, 2 hours before laparotomy. Anesthetized rats were subjected to laparotomy and total hepatic ischemia (30 minutes) induced by by clamping of portal triad. Control group underwent peritoneal puncture, two hours before the sham operation (laparotomy only). At the end of ischemia (G1 and G2), the liver was reperfused for 60 minutes. Following reperfusion blood samples were collected for evaluation of alanine aminotransferase (ALT) and lactate dehydrogenase (LDH) levels. Liver (medium lobe) was removed for immunohistochemistry study with antibody for Caspase-3. RESULTS: It was found a significant decrease (p<0.05) of ALT levels (270.6 +40.8 vs 83.3 +5.5 - p <0.05), LDH (2079.0 +262.4 vs. 206.6 +16.2 - p <0.05) and Caspase-3 expression (6.72 +1.35 vs. 2.19 +1.14, p <0.05) in rats subjected to I / R, comparing the group treated with L-Ala -Gln with G-2. Also, the ALT level was significantly lower (P<0.05) in G-1 and G-2 groups than in G-3 (control group). CONCLUSION: L-Ala-Gln preconditioning in rats submitted to hepatic I/R significantly reduces ALT, LDH and Caspase-3 expression, suggesting hepatic protection.

P0538HYPERIN PREVENTS ISCHEMIC/REPERFUSION AKI VIA STK40

2020 ◽  
Vol 35 (Supplement_3) ◽  
Author(s):  
Yan Xu ◽  
Yue Zhang
Keyword(s):  
Caspase 3 ◽  
Ischemia Reperfusion ◽  
Normal Group ◽  
Sham Operation ◽  
In Vitro Experiments ◽  
Rt Pcr ◽  
Sham Operation Group ◽  
Tnf Α ◽  
Operation Group

Abstract Background and Aims Ischemia-reperfusion injury (IRI) is the outcome of an inflammatory process and tubular cell death that is triggered by undergoing a transient reduction or cessation of blood flow and following by reperfusion. Unresolved IRI can contribute to chronic kidney disease even death. Our aims is to investigate the protective effect of hyperin on ischemia-reperfusion renal injury (IRI) and its possible mechanism. Method ① The transcriptome chip data of multiple IRI models were selected from the NCBI GEO DateSets database and a number of key proteins that could participate in IRI were screened out (the fold increase was greater than 2 fold and was statistically significant). Network and transcript binding motif analysis was performed to determine the best binding protein. ② C57BL / 6J mice were selected and randomly divided into normal group, sham operation group, IRI group (bilateral renal pedicle clamping for 45min), hyperin + IRI group (50mg / kg.d per day, 7 days before surgery ), DMSO + IRI group (7 days before the operation, the same amount of DMSO was administered to the stomach every day, and the operation was the same as AKI), with 6 rats in each group. Renal tissue and blood were collected 24 hours after operation for testing. ③ In vitro experiments, human proximal tubule epithelial cells (HK-2) were selected and divided into hypoxia 3, 6, 9, 12, 24, 36, and 48h for reoxygenation of 1, 3, and 6h respectively. Relevant indicators for RT-PCR detection were determined Optimal hypoxia time. The drug safe concentration was selected according to 0, 5, 10, 25, 50, 100, 200, 400 μg / ml hyperin pre-treatment for 12 hours, and the CCK8 reagent was added for 2 hours to measure the absorbance at 450 nm. The cells were randomly divided into normal group, hypoxia group, hypoxia + DMSO group, hypoxia + hyperin group, and related indexes were detected by RT-PCR and Western Blot. ④ Obtain the tertiary structure of the protein and the three-dimensional structure of the hyperin molecule from the RCSB Protein Data Bank website and the PubChem compound database, and use molecular docking technology to determine the proteins that can bind to hyperin using autodock software and analyze their binding ability. Results Bioinformatics analysis suggested that STK40 protein is one of the key factors of IRI and may be a target for preventing and treating diseases. In vivo experiments showed that compared with the normal group and the sham operation group, the levels of serum creatinine, blood urea nitrogen, and kim-1 in rats were significantly increased after AKI, and HE staining of pathological sections showed an increase in renal tubular injury scores. Significantly decreased (P&lt;0.05); RT-PCR results showed that kim-1, caspase-3, NF-κB, IL-6, TNF-α increased significantly after AKI, STK40, Bcl2 / BAX decreased, and the above after hyperin The indicators changed in opposite directions (P &lt;0.05). In vitro experiments: The best time for hypoxia is 24h hypoxia + 1h reoxygenation; compared with the control group, the drug concentration is &lt;100 μg / mL and the cell proliferation activity rate is&gt; 90%, so the hyperin concentration was selected as 50 μg / mL (P &lt; 0.05); RT-PCR results showed that Hif1-α, caspase-3, NF-κB, IL-6, TNF-α significantly increased, and STK40, Bcl2 / BAX decreased compared with the normal group. After administration of hyperin, the above indexes changed in opposite directions (P &lt;0.05). Conclusion In this study, using molecular docking technology and constructing IRI mice model, it was confirmed that hyperin can reduce IRI and exert a protective effect on IRI by inhibiting STK40 expression.

Study on facial nerve activation pathway based on nanometric magnetic beads

2020 ◽  
Vol 10 (11) ◽  
pp. 1808-1815
Author(s):  
Yang Yuan ◽  
Liheng Li ◽  
Wenwen Liu ◽  
Hong Liu ◽  
Qijun Fan ◽  
...  
Keyword(s):  
Facial Nerve ◽  
Western Blotting ◽  
Magnetic Beads ◽  
Morphological Changes ◽  
Control Group ◽  
Rt Pcr ◽  
Gene Target ◽  
Facial Nerve Surgery ◽  
Operation Group ◽  
Nerve Surgery

This study was conducted to investigate the mechanism of facial nerve recovery through Toll-like receptor 2 (TLR2)/nuclear factor kappa B (NF-κB) after facial nerve trunk damage. Facial nerve surgery was performed on rats to establish a paralyzed animal model with or without intraperitoneal (i.p.) injection of Pam3CSK4 (TLR2 agonist). All nucleic acids were extracted using nanometer magnetic beads. At 1, 4, 7, 10, 13, and 16 days after the surgery, the score of the blink reflex, the motion of vibrissae, and the motion of nose were analyzed to assess the function of facial nerves. Evaluated using transmission electron microscopy (TEM) and immunofluorescence staining (IF), Morphological changes in facial nerve and brainstem were analyzed. The expressions of TLR2 and NF-κB p65 in the brainstem were detected by quantitative Reverse Transcription-Polymerase Chain Reaction (RT-PCR), western blotting, and IF. TEM revealed that the facial nerve fiber diameter was shortened, and the organelle was swollen and demyelinated in the operation group. The IF results revealed that the expressions of NF-κB p65 and TLR2 in the brainstem were increased significantly in the operation group together with the control group. RT-PCR demonstrated that the mRNA expressions of TLR2 and NF-κB p65 in the brainstem were low in the control group, as well as in the sham group, whereas there were significant increases in the mRNA expressions of TLR2 and NF-κB p65 in the operation group. Western blotting revealed a significant increase in the expression of TLR2 protein after the surgery compared to that in the control group. After subjecting the rats to facial nerve surgery and i.p. injection of Pam3CSK4, the protein expressions of TLR2 and NF-κB p65 were upregulated dramatically in the Pam3CSK4-operation group compared to that in the control group and were also significantly increased than those in the operation group. Both facial nerve dysfunction caused due to facial nerve surgery (exacerbated by co-treatment with Pam3CSK4) and the expressions of NF-κB p65 and TLR2 were increased significantly. TLR2 might be an innovative gene target and a novel option for the clinical applications of facial paralysis immunotherapy.

scholarly journals Effect of Curcumol on the Fenestrae of Liver Sinusoidal Endothelial Cells Based on NF-κB Signaling Pathway

2020 ◽  
Vol 2020 ◽  
pp. 1-9
Author(s):  
Yang Zheng ◽  
Jiahui Wang ◽  
Jiaru Wang ◽  
Haiyuan Xie ◽  
Tiejian Zhao
Keyword(s):  
Electron Microscopy ◽  
Scanning Electron Microscopy ◽  
Signaling Pathway ◽  
Intervention Group ◽  
Positive Control ◽  
Positive Control Group ◽  
Control Group ◽  
Rt Pcr ◽  
Liver Sinusoidal Endothelial Cells ◽  
Scanning Electron

Objective. To study the effect of curcumol on liver sinusoidal endothelial cells (LSECs) and to analyze the mechanism of antihepatic fibrosis. Methods. The effects of drug intervention on cell proliferation rates were detected by MTT assay. The expression of NF-κB was detected by RT-PCR and WB. The NF-κB expression and entry into the nucleus were detected by immunofluorescence; scanning electron microscopy was used to observe the changes of LSECs fenestrae. Results. MTT results showed that the interference of cell proliferation in each group was small. RT-PCR showed that the expression of NF-κB in the curcumol intervention group was significantly lower than that in the positive control group (P<0.05). The WB detection found that, in the curcumol intervention group, the expression of pNF-κB in the NF-κB signaling pathway was significantly lower than that in the positive control group (P<0.05). Scanning electron microscopy showed that the LSEC fenestrae were significantly improved compared with the positive control group. Conclusion. Curcumol may be one of the mechanisms of antihepatic fibrosis by inhibiting the activity of the NF-κB signaling pathway and increasing the fenestrae of LSECs.

Study on the Effects of Rapamycin Reverse AMN107 resistance on k562/AO2 Cells

Blood ◽  
2011 ◽  
Vol 118 (21) ◽  
pp. 5017-5017
Author(s):  
Jishi Wang ◽  
Chang Yang ◽  
Cheng Chen ◽  
Qin Fang
Keyword(s):  
Western Blot ◽  
Cell Line ◽  
Chronic Myelogenous Leukemia ◽  
Copy Number ◽  
Caspase 3 ◽  
Annexin V ◽  
Myelogenous Leukemia ◽  
Control Group ◽  
Rt Pcr ◽  
Regulation Of Expression

Abstract Abstract 5017 Aim Rapamycin(RAPA) is the inhibitor of m-TOR. The activation of m-TOR results in changes in multiple cellular processes, eg, catabolism, anabolism, proliferation, growth and apoptosis. Rapamycin, which directly inhibits the Akt downstream molecule mammalian target of rapamycin (mTOR), effectively inhibits survival and proliferation in the development of BCR- ABL–induced chronic myelogenous leukemia(CML) from PTENfl/fl. Nilotinib (AMN107), a BCR-ABL tyrosine kinase inhibitor, was developed to surmount resistance or intolerance to imatinib in patients with Philadelphia positive chronic myelogenous leukemia. We aim to introduce the CML cell line (k562/A02) resistant to AMN107 and investigate the effect of RAPA reverse AMN107 resistance on k562/A02 cell line. Method K562/A02 be added in the culture medium final concentration 1μmol/ L of AMN107 drugs a week before the withdrawal. For all experiments, cells were on the logarithmic phase.K562/A02 cells were treated with 400nmol/L rapamycin(400nmol/L RAPA group),100nmol/L rapamycin(100nmol/L RAPA group), 20μmol/L nilotinib(20μmol/L AMN107 group), 400nmol/L rapamycin +20μmol/L nilotinib (400nmol/L RAPA+20μmol/L AMN107 group), 100nmol/L rapamycin +20μmol/L nilotinib (100nmol/L RAPA+20μmol/L AMN107 group). Tetrazolium—based colorimetric assay (MTT) was used to detect the inhibiton effect of cell growth. The expression of Survivin, m-TOR, Bcl-2, and Caspase-3 were detected by western blot; Cell apoptosis was inspected by Annexin V/PI double staining method; The expression of Survivin, m-TOR, Bcl-2 and Caspase-3 in mRNA level was determined using reverse transcriptase—polymerase chain reaction (RT-PCR). BCR-ABL gene was detected by Real Time-PCR when K562/AO2 cells in different groups were treated by drugs for 48h. Result K562/A02 cells were successfully introduced to resistant toAMN107. Combination of 400 and 100 nmol/L RAPA with AMN107 could significantly enhance the sensitivity of k562/A02 to AMN107. The reverse factor was 1.97 and 2.73 fold respectively. RT-PCR and Western blot method indicated that the expression of Survivin and Bcl-2 was increased by RAPA+AMN107 group, but the MDR1 and Caspase-3 was decreased. The expression of BCR-ABL fusion gene in100 nmol/L RAPA group was higher than the blank groups, the copy number is 5.6×10−2. But the copy number in 400nmol/L RAPA +20μmol/L AMN107 group is1.64×10−2 that is lowest. Annexin V/PI showed that the apoptosis rates were (8.21±0.05)%, (32.07±0.03)%, (28.88±0.11)%, (16.57±0.05)%, (38.49±0.04)%, and(42.31±0.07)% respectively in control group, AMN107 group, 400nmol/L RAPA group, 100nmol/L RAPA group, 100nmol/L RAPA +20μmol/L AMN107 group, and 400nmol/L RAPA +20μmol/L AMN107 group. Conclusion We successfully introduced K562/A02 resistant toAMN107. Moreover, RAPA can enhance the sensitivity of AMN107 resistant 562/A02 cells. Combined of RAPA and AMN107 has significant synergistic growth inhibiting effect and apoptosis inducing effect on AMN107-resistant K562/A02 cells. The mechanism may be related at down-regulation of expression MDR1 and up-regulation of expression Survivin. Disclosures: No relevant conflicts of interest to declare.

Scanning Electron Microscopy and Roughness Study of Dental Composite Degradation

2012 ◽  
Vol 18 (2) ◽  
pp. 289-294 ◽  
Author(s):  
Luís Eduardo Silva Soares ◽  
Louise Ribeiro Cortez ◽  
Raquel de Oliveira Zarur ◽  
Airton Abrahão Martin
Keyword(s):  
Electron Microscopy ◽  
Scanning Electron Microscopy ◽  
Surface Roughness ◽  
Composite Resin ◽  
Morphological Changes ◽  
Soft Drink ◽  
Dental Composite ◽  
Control Group ◽  
Significant Difference ◽  
Scanning Electron

AbstractOur aim was to test the hypothesis that the use of mouthwashes, consumption of soft drinks, as well as the type of light curing unit (LCU), would change the surface roughness (Ra) and morphology of a nanofilled composite resin (Z350® 3M ESPE). Samples (80) were divided into eight groups: Halogen LCU, group 1, saliva (control); group 2, Pepsi Twist®; group 3, Listerine®; group 4, Colgate Plax®; LED LCU, group 5, saliva; group 6, Pepsi Twist®; group 7, Listerine®; group 8, Colgate Plax®. Ra values were measured at baseline, and after 7 and 14 days. One specimen of each group was prepared for scanning electron microscopy analysis after 14 days. The data were subjected to multifactor analysis of variance at a 95% confidence followed by Tukey's honestly significant difference post-hoc test. All the treatments resulted in morphological changes in composite resin surface, and the most significant change was in Pepsi Twist® groups. The samples of G6 had the greatest increase in Ra. The immersion of nanofilled resin in mouthwashes with alcohol and soft drink increases the surface roughness. Polymerization by halogen LCU (reduced light intensity) associated with alcohol contained mouthwash resulted in significant roughness on the composite.

Cisplatin Nano-Liposomes Promoting Apoptosis of Retinoblastoma Cells Both In Vivo and In Vitro

2020 ◽  
Vol 12 (4) ◽  
pp. 536-542
Author(s):  
Lijuan Zhao ◽  
Fei Wang ◽  
Wei Fan
Keyword(s):  
Protein Expression ◽  
Nude Mice ◽  
Caspase 3 ◽  
Mrna Level ◽  
Control Group ◽  
Mrna Levels ◽  
Bax Protein ◽  
Experimental Group

This study was established to investigate the effects of cisplatin nano-liposomes on the apoptosis of the human retinoblastoma (RB) cell line Y79 in vitro and in vivo. Y79 cells were cultured and then exposed to Annexin V/PI to test their apoptosis, tested with the Caspase-3 activity detection kit to examine the change in activity of Caspase-3, and subjected to western blotting to test Bcl-2 and Bax protein expression. Y79-cell-transplanted tumor model in nude mice was also established and divided into three groups, with five nude mice in each. Cisplatin nano-liposomes were applied to the experimental group, cisplatin was injected into the control group, while saline was administered to the blank group, after which the nude mice were killed and the tumor was removed. Tumor volumes and weights in the three groups were compared. Nucleic acid extraction from magnetic beads was adopted to extract DNA, RT-PCR was employed to test Bcl-2 and Bax mRNA levels in tumor tissues, and in situ cell death assay kit was applied to test apoptotic cells. In comparison to the cisplatin solution and DMSO groups, the cisplatin liposome group showed higher Y79 apoptotic rate, Caspase-3 activity, and Bax protein expression, and lower Bcl-2 protein expression (all P < 0 05). In comparison with the control and blank groups, the experimental group showed lower tumor volume, weight, and Bcl-2 mRNA level of nude mice. In addition, in comparison with the control group, the experimental group showed higher cellular apoptotic rate and Bax mRNA level. In terms of the clinical effects of cisplatin nano-liposomes on a tumor transplant in nude mice with cervical cancer, they were shown to promote tumor apoptosis.

scholarly journals Anticancer Activity of Polysaccharides Produced from Glycerol and Crude Glycerol by an Endophytic Fungus Chaetomium globosum CGMCC 6882 on Human Lung Cancer A549 Cells

Biomolecules ◽  
2018 ◽  
Vol 8 (4) ◽  
pp. 171 ◽  
Author(s):  
Zichao Wang ◽  
Peizhang Chen ◽  
Ning Tao ◽  
Huiru Zhang ◽  
Ruifang Li ◽  
...  
Keyword(s):  
Crude Glycerol ◽  
Caspase 3 ◽  
A549 Cells ◽  
Biodiesel Production ◽  
Chaetomium Globosum ◽  
Human Lung Cancer ◽  
Control Group ◽  
Rt Pcr ◽  
Caspase 9 ◽  
Molecular Weights

Two polysaccharides were produced by Chaetomium globosum CGMCC 6882 from glycerol (GCP-1) and crude glycerol (GCP-2). Chemical characteristics results showed GCP-1 and GCP-2 were similar polysaccharides, but the molecular weights of GCP-1 and GCP-2 were 5.340 × 104 Da and 3.105 × 104 Da, respectively. Viabilities of A549 cells after treatment with GCP-1 and GCP-2 were 49% and 39% compared to the control group. Meanwhile, flow cytometry results indicated that GCP-1 and GCP-2 could induce 17.79% and 24.28% of A549 cells to apoptosis with 200 μg/mL concentration treated for 24 h. RT-PCR results suggested that GCP-1 and GCP-2 could be used as potential and effective apoptosis inducers on A549 cells by increasing BAX, CASPASE-3, CASPASE-9, TIMP-1, TIMP-2 expression and decreasing BCL-2 expression. This research provided an innovative approach to using a byproduct of biodiesel production (crude glycerol) to produce polysaccharides of potential medicinal benefit.

scholarly journals Intervention and Mechanism of Dexmedetomidine on A549 Cells Injured by Hypoxia/Reoxygenation

2019 ◽  
Vol 9 (3) ◽  
pp. 99-106
Author(s):  
Hui Gao ◽  
Ziyin Luo ◽  
Zhuolin Zhou ◽  
Yiming Wu ◽  
Wantie Wang
Keyword(s):  
Western Blot ◽  
Caspase 3 ◽  
Morphological Changes ◽  
Intervention Group ◽  
A549 Cells ◽  
Rt Pcr ◽  
Caspase 12 ◽  
Injury Group ◽  
Apoptosis Index ◽  
Hypoxia Reoxygenation

Objective: To investigate the effect and mechanism of dexmedetomidine (DEX) on hypoxia /reoxygenation (H/R) injury of A549 cells. Methods: A549 cells were cultured and randomly divided into four groups (n=10): Normoxic group;DEX group; H/R injury group;H/R injury+DEX intervention group. Observe the morphological changes of cells; Cell viability was detected by cck-8 assay. TUNEL assay was used to detect apoptosis index (AI).Expressions of GRP78, CHOP, JNK, caspase-12, caspase-3 proteins and mRNA were detected by Western Blot and RT-PCR;Detect the activity of caspase-3. Results: Compared with the H group, the OD value and AI value in the HD group were significantly up-regulated, apoptotic cells were significantly decreased, the expressions of CHOP, caspase-12, p-JNK and caspase-3 proteins and mRNA were significantly decreased, the GRP78 protein and mRNA increased, and the caspase-3 activity was significantly decreased, the differences were statistically significant (P<0.01). Conclusion: Dexmedetomidine has a protective effect on A549 cells after H/R injury, which may be related to its inhibition of apoptosis induced by excessive endoplasmic reticulm stress. Keywords: Dexmedetomidine; Hypoxia/Reoxygenationinjury;Apoptosis;Endoplasmicreticulm stress.

scholarly journals Chinese Medicine FTZ Recipe Protects against High-Glucose-Induced Beta Cell Injury through Alleviating Oxidative Stress

2019 ◽  
Vol 2019 ◽  
pp. 1-14 ◽  
Author(s):  
Weijian Bei ◽  
Yujiao Wang ◽  
Jianmei Chen ◽  
Jingjing Zhang ◽  
Lexun Wang ◽  
...  
Keyword(s):  
Oxidative Stress ◽  
Protein Expression ◽  
High Glucose ◽  
Caspase 3 ◽  
Fasting Blood Glucose ◽  
Lipid Peroxide ◽  
Control Group ◽  
Cell Dysfunction ◽  
Bax Protein ◽  
Diabetic Model

Objective. To investigate the effect of FTZ on high-glucose-induced oxidative stress and underlying mechanisms. Methods. We used a β cell dysfunction and diabetes model that was induced in rats fed a high-fat high-sugar diet (HFHSD) for 6 weeks and injected once with 35 mg/kg streptozocin (STZ). Then, 3 and 6 g/kg of FTZ were administered by gavage for 8 weeks. In addition, an ex vivo model of oxidative stress was induced by stimulating INS-1 cells with 25 mmol/L glucose for 48 h. Result. The levels of fasting blood glucose (FBG) in diabetic model rats were obviously higher than those in the normal group; furthermore with reduced levels of β cells, catalase (CAT), superoxide dismutase (SOD), and Bcl-2 increased lipid peroxide malondialdehyde (MDA) and caspase-3 in the pancreatic tissue of the diabetic model rats. Afterward, the cells were incubated with FTZ-containing serum and edaravone. The 25 mmol/L glucose-induced SOD reduction increased MDA and intracellular ROS. The protein expression level of Mn-SOD and CAT in the model group decreased significantly compared with that in the control group. Conclusion. FTZ treatment significantly improved the alteration in the level of SOD, CAT, Bcl-2, caspase-3, and MDA coupled with β cell dysfunction in diabetic rats. Oxidative stress in INS-1 cells was closely associated with a higher rate of apoptosis, increased production of ROS and MDA, enhanced Bax expression, and caspase-3, -9 activities and markedly decreased protein expression of Mn-SOD and CAT. FTZ-containing serum incubation notably reversed the high-glucose-evoked increase in cell apoptosis, production of ROS and MDA, and Bax protein levels. Furthermore, FTZ stimulation upregulated the expression levels of several genes, including Mn-SOD, CAT, and Bcl-2/Bcl-xl. In addition, FTZ decreased the intracellular activity of caspase-3, -9 in INS-1 cells. FTZ protected β-cells from oxidative stress induced by high glucose in vivo and in vitro. The beneficial effect of FTZ was closely associated with a decrease in the activity of caspase-3, -9 and intracellular production of ROS, MDA, and Bax coupled with an increase in the expression of Mn-SOD, CAT, and Bcl-2/Bcl-xl.

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