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Objective:To evaluate the effect of chicoric acid on oxidative stress during myocardial injury in sepsis rats and the relationship with nuclear factor E2-related factor 2 (Nrf2) signaling pathway.Methods:Forty healthy male Sprague-Dawley rats, aged 8-12 weeks, weighing 220-250 g, were divided into 5 groups ( n=8 each) using a random number table method: control group (group C), lipopolysaccharide (LPS) group (group LPS), LPS+ chicoric acid group (group LPS+ CA), LPS+ Nrf2 inhibitor ML385 group (group LPS+ ML) and LPS+ chicoric acid+ ML385 group (group LPS+ CA+ ML). LPS 15 mg/kg was intraperitoneally injected to induce sepsis.Immediately after intraperitoneal injection of LPS, chicoric acid 10 mg/kg or ML385 15 mg/kg (in dimethyl sulfoxide) was intraperitoneally injected in group LPS+ CA and group LPS+ ML, respectively, and ML385 15 mg/kg and chicoric acid 10 mg/kg were intraperitoneally injected in LPS+ CA+ ML group.The equal volume of dimethyl sulfoxide was given instead in group C. At 48 h after establishment of the model, blood samples were collected from the aorta for measurement of concentration of serum interleukin-6 (IL-6) and the activities of lactate dehydrogenase (LDH) and creatine kinase isoenzyme (CK-MB) (by enzyme-linked immunosorbent assay). The animals were then sacrificed, and myocardial tissues were obtained for microscopic examination of pathological changes (by HE staining), for determination of activities of glutathione peroxidase (GSH-Px) and superoxide dismutase (SOD) and contents of reactive oxygen species(ROS) and iron (by colorimetry), for calculation of the ratio of oxidized nicotinamide adenine 2 nucleotides to reduced nicotinamide adenine 2 nucleotides (NAD + /NADH), and for detection of the expression of Nrf2, NADPH quinone oxidoreductase 1 (NQO1), glutathione peroxidase 4 (GPX4) and nicotinamide adenine dinucleotide phosphate oxidase 1 (NOX1) (by Western blot). Results:Compared with C group, the activities of serum LDH and CK-MB and concentration of IL-6 were significantly increased, the contents of ROS and iron and the ratio of NAD + /NADH were increased, activities of GSH-Px and SOD were decreased, expression of Nrf2, NQO1 and GPX4 was down-regulated, and NOX1 expression was up-regulated in the other four groups ( P<0.05). Compared with group LPS, the activities of serum LDH and CK-MB and concentration of IL-6 were significantly decreased, the contents of ROS and iron and the ratio of NAD + /NADH were decreased, activities of GSH-Px and SOD were increased, expression of Nrf2, NQO1 and GPX4 was up-regulated, NOX1 expression was down-regulated ( P<0.05), and the pathological changes of cardiomyocytes were significantly reduced in group LPS+ CA, and the activities of serum LDH and CK-MB and concentration of IL-6 were significantly increased, the ratio of NAD + /NADH were increased, activities of GSH-Px and SOD were decreased, expression of Nrf2, NQO1 and GPX4 was down-regulated, NOX1 expression was up-regulated ( P<0.05), and the pathological changes of cardiomyocytes were accentuated in group LPS+ ML.Compared with group LPS+ CA, the activities of serum LDH and CK-MB and concentration of IL-6 were significantly increased, the contents of ROS and iron and the ratio of NAD + /NADH were increased, activities of GSH-Px and SOD were decreased, expression of Nrf2, NQO1 and GPX4 was down-regulated, NOX1 expression was up-regulated ( P<0.05), and the pathological changes of cardiomyocytes were accentuated in group LPS+ CA+ ML. Conclusion:The mechanism by which chicoric acid reduces myocardial injury in sepsis rats may be related to activating Nrf2 signaling pathway and inhibiting oxidative stress.
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Objective To investigate the protective effect of salvianolic acid A (SAA) on permanent focal cerebral ischemia in rats and its possible mechanisms. Methods Fifty-four adult male Sprague-Daw ley rats w ere randomly divided into a sham operation group, a cerebral ischemia group, and a SAA group ( n =18 in each group). A model of permanent middle cerebral artery occlusion w as induced by the intraluminal suture method.At 0 h and 6 h after modeling, the rats of the SAA groups w ere intraperitonealy injected SAA (3 mg/kg). The other groups w ere injected equal volume of saline. At 24 h after modeling, the neurological deficit scores w ere performed. 2,3,5-Triphenyl tetrazolium chloride (TTC) staining w as used to detect cerebral infarction volume. TUNEL staining w as used to detect cel apoptosis. Both immunohistochemical staining and Western blotting w ere used to detect the expressions of Wnt3a, β-catenin, and phosphor-glycogen synthase-kinase-3β(p-GSK-3β) in the ischemic cortex. Results The neurological deficit scores show ed that no neurological deficits w ere observed in the sham operation group (score 0). The neurological deficit score in the SAA group (median and interquartile range) w as significantly low er than that in the cerebral ischemia group (3 [2-3] vs.4 [3-5]; Z = -2.679, P =0.007). No infarcts w ere observed in the sham operation group. The infarct volume in the SAA group w as reduced significantly compared w ith the cerebral ischemia group (79.038 ±10.665 mm 3 vs.212.702 ±8.029 mm 3; t = 24.525, P < 0.001). Very few positive cels w ere observed in the sham operation group. The numbers of TUNEL -positive cels in the SAA group and the cerebral ischemia group w ere 29.667 ±1.366/HP and 63.333 ±0.894/HP, respectively. The former w as significantly less than the latter ( t = 14.115, P < 0.001). Immunohistochemical staining show ed that the number of Wnt3a positive cels in the sham operation group, the cerebral ischemia group, and the SAA group w ere 35.500 ±2.572/HP, 18.056 ±3.765/HP, and 29.000 ±2.376/HP, respectively. There w ere significant differences among the 3 groups ( F = 115.972, P < 0.001), and those in the SAA group w ere significantly more than the cerebral ischemia group ( P < 0.01). The numbers of p-GSK-3βpositive cels in the sham operation group, the model group, and the SAA group w ere 7.944 ±2.127/HP, 37.444 ±3.434/HP, and 11.222 ±1.734/HP, respectively. There w ere significant differences among the three groups (F =730.580, P < 0.001), and those in the SAA group w ere significantly less than the cerebral ischemia group ( P < 0.01). The numbers of β-catenin positive cels in the sham operation group, the cerebral ischemia group, and the SAA group w ere 26.722 ±26.722/HP, 16.556 ±1.854/HP, and 21.333 ± 1.940/HP, respectively. There w ere also significant differences among the 3 groups ( F < 33.385, P <0.01), and those in the SAA group w ere significantly more than the cerebral ischemia group ( P < 0.01). Western blot analysis show ed that Wnt3a expression levels in the sham operated group, the cerebral ischemia group, and the SAA group w ere 1.000 ±0.190, 0.800 ±0.185, and 1.198 ±0.262, respectively. There w ere significant differences among 3 groups ( F = 9.621, P < 0.001), and those in the SAA group w ere significantly higher than the cerebral ischemia group ( P < 0.01). The p-GSK-3βexpression levels in the sham operation group, the cerebral ischemia group, and the SAA group w ere 0.650 ±0.150, 1.290 ± 0.250, and 1.190 ±0.250, respectively. There w ere also significant differences among the 3 groups ( F =19.668, P < 0.001), and those in the SAA group w ere significantly higher than the cerebral ischemia group (P <0.01). The β-catenin expression levels in the sham operation group, the cerebral ischemia group, and the SAA group w ere 1.200 ±0.210, 0.500 ±0.120, and 1.100 ±0.220, respectively. There w ere significant differences among the 3 groups ( F = 33.385, P < 0.001), and those in the SAA group w ere significantly higher than the cerebral ischemia group ( P < 0.01). Conclusions SAA has certain protective effect on permanent cerebral ischemia injury in rats. Its mechanism may be associated w ith the up -regulation of the expression of Wnt3a and β-catenin and the dow n-regulation of the expression of p-GSK-3β.
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Objective: To assess the effects of caffeic acid (CA) on MPP + induced cerebellar granule neurons (CGNs) apoptosis. Methods: CGNs were pretreated with caffeic acid at 55, 110 and 220 ?mol/L for 6 h, then treated with 100 ?mol/L MPP + for 24 h (concentration effect relationship). In addition CGNs were pretreated with caffeic acid at 110 ?mol/L for 0 h, 6 h, 12 h, and 24 h, respectively, then treated with 100 ?mol/L MPP + for 24 h (time response relationship). Besides, after treatment with MPP + for 24 h, CGNs were incubated with caffeic acid at 55, 110 and 220 ?mol/L,respectively. Cell viability was determined by 3 (4,5 dimethylthiazol 2 yl) 2,5 diphenyltetrazolium bromide (MTT) assay and caspase 3 activity was assayed by caspase 3 fluorometric assay kit. Results: MTT assay revealed that caffeic acid significantly inhibited cell viability decrease induced by MPP +, and caspase 3 fluorometric assay showed that caffeic acid efficiently suppressed caspase 3 activation in CGNs induced by MPP +. Conclusion: Caffeic acid (CA) can significantly protect CGNs from apoptosis induced by MPP + and may provide a useful therapeutic strategy for the treatment of Parkinson's disease.
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AIM:To evaluate the antitumor effect of caffeic acid Ge on U14 tumor bearing mice.METHODS:The tumor inhibitory ratios of caffeic acid Ge on the growth of U14 in mice was observed.Apoptosis morphological transformation of U14 cells induced by caffeic acid Ge was detected by electronic scan microscope and MG-P staining.Alteration of cell cycle was analyzed by flow cytometry.Apoptosis-related protein levels of Bax and Bcl-2 were determined by immunity histochemistry technology.MTT assay was applied to study the antitumor activities of caffeic acid Ge in U14 cell lines in vitro.RESULTS:Tumor inhibitory rates in caffeic-acid Ge groups were 38.50%,47.17% and 64.02%(from low dose to high dose)(P