ABSTRACT
AIM: To investigate the effect and mechanism of matrix metalloproteinase(MMPs)inhibitor AG3340 on the migration and invasion ability of retinal pigment epithelial cells-19(ARPE-19)cultured in high glucose(CHG). METHODS: ARPE-19 cells cultured in vitro were divided into four groups: Control group, the glucose at the concentration of 5.6mmol/L in DMEM/F12 medium; HG group, the glucose at the concentration of 30mmol/L was cultured with DMEM/F12 medium; HG+AG3340 group, the cells were pretreated with AG3340 for 12h, and then cultured in DMEM/F12 medium containing 30mmol/L glucose; The mannitol(MA)group, cultured with DMEM/F12 medium of 5.6mmol/L glucose and 24.4mmol/L mannitol, which used as hypertonic control group. The migration ability of cells was detected by wound healing assay, the invasion ability of cells was detected by Transwell assay, and the relative expression levels of MMP-9, MMP-2, fibronectin and collagen Ⅳ were detected by Western blot.RESULTS: The results of wound healing assay showed that compared with the Control group, the cell migration rate of scratching after 24h and 48h in the HG group was significantly increased(all P<0.001).After pretreated by AG3340, the cell migration rate was significantly lower than that in the HG group(all P<0.01).Transwell assay showed that compared with the Control group, the number of cell invasion in the HG group was significantly higher than that in the Control group(all P<0.001). After pretreated by AG3340, the number of cell invasion was decreased than the HG group(all P<0.01). Western blot results showed that compared with the Control group, the relative expression levels of MMP-9 and MMP-2 of the cells in the HG group were increased, and the relative expression levels of Fibronectin and Collagen Ⅳ were decreased(all P<0.001). Compared with the HG group, the relative expression levels of MMP-9 and MMP-2 protein in AG3340 pretreatment group were decreased, and the relative expression levels of Fibronectin and Collagen Ⅳ were increased(all P<0.05). CONCLUSION: High glucose induced ARPE-19 cells with enhanced migration and invasion ability, and AG3340 partially reversed this effect, which was related to the inhibition of MMP-9 and MMP-2 expression and the stability of extra-cellular matrix components.
ABSTRACT
?AIM:To analyze the morphological changes of macular soft drusen and drusenoid pigmental epithulium detachment ( DPED ) after subthreshold micropulse laser treatment ( SMLT) .?METHODS: Fourteen patients ( 20 affected eyes ) with soft drusen and DPED clinically confirmed from August 2016 to October 2018, were included in this study. 577 nm yellow laser of SMLT was applied for soft drusen and DPED. The changes of soft drusen and DPED in best corrected visual acuity ( BCVA ) ( LogMAR ) and height, diameter and cross-sessional area according to fundus autofluorescence and SD - OCT examinations were observed after SMLT.?RESULTS: BCVA was not significant difference after treatment of soft drusen (P=0.260), and the DPED (P=0. 736 ) than that of the baseline. Compared with the baseline values, the height and cross-sessional area of soft drusen were reduced at the 6mo after treatment ( P=0. 008, P=0.034) . Compared with the baseline values, the differences were not statistically significant in height, diameter and cross - sectional area of DPED after treatment.?CONCLUSION: BCVA was not reduced for drusen and DPED after SMLT, however, the height and cross -sessional area of soft drusen was reduced compared with those before treatment, and the differences were not statistically significant in height, diameter and cross -sectional area of DPED before and after treatment. The results indicated that SMLT was effective for soft drusen, but was not effective for short-term treatment of DPED. SMLT caused no damage to the visual acuity in treatment of soft drusen and DPED, but prospective, controlled, large sample and long-term follow-up studies should be required.
ABSTRACT
AIM:To investigate the effect of ischemic postconditioning ( IPC) on autophagy induced by focal cerebral ischemia reperfusion ( I/R) in rats.METHODS:Healthy male SD rats were assigned randomly into sham-opera-tion (sham) group, I/R group and IPC group with 10 rats in each group.The rats in sham group were only exposed the right common , internal and external carotid artery surgically .The rats in I/R group were subjected to right middle cerebral artery occlusion (MCAO) by the modified Longa suture method for 2 h followed by 24 h of reperfusion.The rats in IPC group were subjected to MCAO for 2 h followed by reperfusion of the ipsilateral common carotid artery occlusion for 10 s for 5 episodes, and then reperfusion for 24 h.Autophagy was obeserved by transmission electron microscopy (TEM).The pro-tein levels of mammalian target of rapamycin (mTOR), p-mTOR and microtubule associated protein light chain 3 (LC3)-II in brain tissue of the rats were determined by Western blot .Pathological changes of brain tissue were observed by HE staining.RESULTS:The protein levels of mTOR and p-mTOR in IPC group were significantly higher than those in I/R group (P<0.05).The expression of LC3-II in IPC group was significantly lower than that in I/R group (P<0.01).The cerebral infarction area and brain water content in IPC group were significantly lower than those in I /R group (P<0.01). HE staining showed that neurons degeneration and necrosis in IPC group were significantly alleviated compared with I /R group.TEM observation showed that IPC revealed fewer autophagosomes , with much less severe cell damage than that in I/R group.CONCLUSION:IPC reduces brain ischemia reperfusion damage by decreasing autophagy of brain cells , which might be related to the activation of mTOR .
ABSTRACT
AIM:To investigate the effect of ischemic postconditioning ( IPC) on autophagy induced by focal cerebral ischemia reperfusion ( I/R) in rats.METHODS:Healthy male SD rats were assigned randomly into sham-opera-tion (sham) group, I/R group and IPC group with 10 rats in each group.The rats in sham group were only exposed the right common , internal and external carotid artery surgically .The rats in I/R group were subjected to right middle cerebral artery occlusion (MCAO) by the modified Longa suture method for 2 h followed by 24 h of reperfusion.The rats in IPC group were subjected to MCAO for 2 h followed by reperfusion of the ipsilateral common carotid artery occlusion for 10 s for 5 episodes, and then reperfusion for 24 h.Autophagy was obeserved by transmission electron microscopy (TEM).The pro-tein levels of mammalian target of rapamycin (mTOR), p-mTOR and microtubule associated protein light chain 3 (LC3)-II in brain tissue of the rats were determined by Western blot .Pathological changes of brain tissue were observed by HE staining.RESULTS:The protein levels of mTOR and p-mTOR in IPC group were significantly higher than those in I/R group (P<0.05).The expression of LC3-II in IPC group was significantly lower than that in I/R group (P<0.01).The cerebral infarction area and brain water content in IPC group were significantly lower than those in I /R group (P<0.01). HE staining showed that neurons degeneration and necrosis in IPC group were significantly alleviated compared with I /R group.TEM observation showed that IPC revealed fewer autophagosomes , with much less severe cell damage than that in I/R group.CONCLUSION:IPC reduces brain ischemia reperfusion damage by decreasing autophagy of brain cells , which might be related to the activation of mTOR .
ABSTRACT
Ammonium perchlorate (AP), mainly used as solid propellants, was reported to interfere with homeostasis via competitive inhibition of iodide uptake. However, detailed mechanisms remain to be elucidated. In this study, AP was administered at 0, 130, 260 and 520 mg/kg every day to 24 male SD rats for 13 weeks. The concentrations of iodine in urine, serum thyroid hormones levels, total iodine, relative iodine and total protein, and malondialdehyde (MDA), superoxide dismutase (SOD) and catalase (CAT) activity in thyroid tissues were measured, respectively. Our results showed that high-dose perchlorate induced a significant increase in urinary iodine and serum thyroid stimulating hormone (TSH), with a decrease of total iodine and relative iodine content. Meanwhile, free thyroxine (FT4) was decreased and CAT activity was remarkably increased. Particularly, the CAT activity was increased in a dose-dependent manner. These results suggested that CAT might be enhanced to promote the synthesis of iodine, resulting in elevated urinary iodine level. Furthermore, these findings suggested that iodine in the urine and CAT activity in the thyroid might be used as biomarkers for exposure to AP, associated with thyroid hormone indicators such as TSH, FT4.
Subject(s)
Animals , Male , Analysis of Variance , Catalase , Metabolism , Dose-Response Relationship, Drug , Homeostasis , Iodine , Metabolism , Urine , Malondialdehyde , Metabolism , Perchlorates , Pharmacology , Quaternary Ammonium Compounds , Pharmacology , Radioimmunoassay , Rats, Sprague-Dawley , Superoxide Dismutase , Metabolism , Thyroid Gland , Metabolism , Thyrotropin , Blood , Thyroxine , Blood , Triiodothyronine , BloodABSTRACT
Ammonium perchlorate (AP), mainly used as solid propellants, was reported to interfere with homeostasis via competitive inhibition of iodide uptake. However, detailed mechanisms remain to be elucidated. In this study, AP was administered at 0, 130, 260 and 520 mg/kg every day to 24 male SD rats for 13 weeks. The concentrations of iodine in urine, serum thyroid hormones levels, total iodine, relative iodine and total protein, and malondialdehyde (MDA), superoxide dismutase (SOD) and catalase (CAT) activity in thyroid tissues were measured, respectively. Our results showed that high-dose perchlorate induced a significant increase in urinary iodine and serum thyroid stimulating hormone (TSH), with a decrease of total iodine and relative iodine content. Meanwhile, free thyroxine (FT4) was decreased and CAT activity was remarkably increased. Particularly, the CAT activity was increased in a dose-dependent manner. These results suggested that CAT might be enhanced to promote the synthesis of iodine, resulting in elevated urinary iodine level. Furthermore, these findings suggested that iodine in the urine and CAT activity in the thyroid might be used as biomarkers for exposure to AP, associated with thyroid hormone indicators such as TSH, FT4.
ABSTRACT
<p><b>OBJECTIVE</b>To investigate the mechanism of thyroid cytotoxicity mechanism of ammonium perchlorate (AP).</p><p><b>METHODS</b>Thyroid cells were cultured in vitro to a certain stage and then exposed to AP (0, 5, 10, 20, 40, and 60 mmol/L) in culture solution; the cultured cells and supernatant were collected. Cell viability was measured by MTT assay; cell apoptosis was determined by flow cytometry; the concentration of thyroglobulin was measured by enzyme-linked immunosorbent assay; the lactate dehydrogenase (LDH) activity, superoxide dismutase (SOD) activity, malondialdehyde (MDA) level, and so on were measured by colorimetry.</p><p><b>RESULTS</b>The cells exposed to 60 mmol/L AP for 12, 24, 48, and 72 h had cell viabilities of 74.93%, 42.26%, 2.66%, and 0.99%, respectively, and the cells exposed to 40 mmol/L AP for 24, 48, and 72 h had cell viabilities of 73.15%, 30.91%, and 3.03%, respectively, all significantly lower than that of the control group (100%)(P < 0.05 or P < 0.01). The overall apoptosis rate of all AP-exposed cells was significantly higher than that of the control group; the cells exposed to 20, 40, and 60 mmol/L AP had early apoptosis rates of 15.70%, 15.84%, and 16.96%, respectively, significantly higher than that of the control group (9.54%)(P < 0.05 or P < 0.01); the cells exposed to 60 mmol/L AP had a late apoptosis rate of 16.54%, significantly higher than that of the control group (6.11%)(P < 0.05 or P < 0.01). The cells exposed to 40 mmol/L AP had a significantly higher LDH activity than the control group (0.70 U/ml vs 0.55 U/ml, P < 0.01). The cells exposed to 5 mmol/L AP had a significantly higher MDA level than the control group (1.08 mmol/L vs 2.36 mmol/L, P < 0.05).</p><p><b>CONCLUSION</b>AP can markedly change the cell morphology and decrease the cell viability of thyroid cells, which may be because AP inhibits cell proliferation, induces cell apoptosis, and destroys cell membranes. However, AP does not result in significant oxidative damage to thyroid cells.</p>
Subject(s)
Humans , Apoptosis , Cell Proliferation , Cell Survival , Cells, Cultured , Oxidative Stress , Perchlorates , Toxicity , Quaternary Ammonium Compounds , Toxicity , Thyroglobulin , Metabolism , Thyroid Gland , Metabolism , PathologyABSTRACT
BACKGROUND & OBJECTIVE: Cerebral hypoxia is known to be involved in many neurodegenerative diseases such as Alzheimer's and cerebrovascular dementia. The present study was designed to investigate the effects of flavonoids from aerial part of Scutellaria baicalensis Georgi (SSF) on potassium cyanide (KCN) -induced hypoxic cytotoxicity in rat pheochromocytoma cell line PC12, and to understand the probable mechanism. METHODS: The rat pheochromocytoma cell line PC12 was subjected to hypoxia by 200 microM KCN for 30 min. The cytotoxicity of KCN was assessed by cell viability assay, morphological observation, lactate dehydrogenase (LDH) release, malondialdehyde (MDA) production, and the activities of superoxide dismutase (SOD) and Na+-K+-ATPase measurements. The effects of SSF on the changes induced by KCN in PC12 cells were detected. RESULTS: Treatment of PC12 cells with 200 micriM KCN for 30 min increased cell death when compared with control, as assayed by MTT reduction, morphological observation and lactate dehydrogenase release measurement. These cell lesions were accompanied by disorders in SOD and Na+-K+-ATPase activities as well as MDA production. In contrast, the PC12 cells pre-treated with SSF for 24 h prior to 200 microM KCN exposure have shown protection against hypoxic toxicity. The KCN - induced decreased cell viability and activities of SOD and Na+-K+-ATPase, as well as increased MDA production were reversed by SSF pre-treatment. INTERPRETATION & CONCLUSION: SSF exerted neuroprotections against KCN - induced hypoxic cytotoxicity in PC12 cells and the probable mechanisms involved free radicals and energy metabolism. Our findings may have implications in future in the treatment of neurodegenerative diseases.
Subject(s)
Animals , Antioxidants/metabolism , Cell Survival/drug effects , Flavonoids/isolation & purification , Humans , Hypoxia, Brain/complications , Neurodegenerative Diseases/drug therapy , Neurons/drug effects , Oxidative Stress/drug effects , PC12 Cells , Potassium Cyanide/toxicity , Rats , Scutellaria baicalensis/chemistryABSTRACT
Objective: To explore the relationship between asymptomatic intracranial arterial stenosis and plasma lipid and uric acid levels in elderly patients with hypertension. Methods: A total of 164 elderly patients with hypertension were selected during the physical examination. Transcranial Doppler (TCD) sonography found that 56 patients had intracranial arterial stenosis (stenotic group) and 108 patients without stenosis (nonstenotic group). In addition, 36 age-and sex-matched healthy controls were selected as control group. Plasma lipid and uric acid levels were detected by automatic biochemistry analyzer in the 3 groups. Results: The levels of plasma total cholesterol (TC), triglyceride (TG), low density lipoprotein cholesterol (LDL-C), and uric acid in the stenotic group were 6.0 ± 1.1 mmol/L, 1.7 ± 0.9 mmol/L, 3.8 ± 0.9 mmol/L, and 437 ± 115 μmol/L, respectively; they were 5.4 ± 1.1 mmol/L, 1.7 ± 1.0 mmol/L, 3.3 ± 0.9 mmol/, and 372 ± 78 μmol/L, respectively in the nonstenstic group; and they were 4.9 ± 0.5 mmol/L, 1.1 ± 0.5 mmol/L, 1.42 ± 0.26 mmol/L, and 324 ± 56 μmol/L, respectively in the control group. There were significant differences between the stenotic and nonstenotic groups with the control group (P<0.001); and there were significant differences between the stenotic group and the nonstenotic group either in the levels of plasma TC, LDL-C, and uric acid (P=0.002, P=0.002, P= 0.000). Conclusion: The increase of the levels of plasma TG, LDL-C and uric acid are the risk factors of intracranial arterial stenosis in elderly patients with hypertension.