ObjectiveTo investigate the effect of α-lipoic acid on the oxidative stress of wound tissues and diabetic wound healing in mice with diabetic feet. MethodsSixty male C57BL/6J mice weighting 200-300 g were randomly divided into model group (control group, n=15), α-lipoic acid-treated model group (n=15), miR-29b mimic group (n=15), and miR-29b mimic negative control group (NC group, n=15). All animals received intraperitoneal injection of streptozocin to establish the diabetic model. Then, a full thickness wound of 5 mm×2 mm in size was created at 4 weeks after modeling. All mice were administrated with high-sugar-fat-diet. At the same day after modeling, α-lipoic acid-treated model group was continuously given intravenous injection of 100 mg/(kg·d) α-lipoic acid for 14 days; miR-29b mimic group and NC group received the tail intravenous injection of lentiviral vector for miR-29b mimic and miR-29b mimic negative control (a total of 2×107 TU), respectively, with the treatment of α-lipoic acid. The wound healing was observed and wound area was measured at 7 and 14 days. The wound tissues were harvested to detect the levels of superoxide dismutase (SOD) and glutathione (GSH) using xanthine oxidase method and 5, 5-dithiobis-2-nitrobenzoic acid staining method at 14 days. At the same day, 7, and 14 days after modeling, the relative miR-29b expression in wound tissues from control and α-lipoic acid-treated model groups was detected by real-time fluorescence quantitative PCR. ResultsAll mice survived to the experiment end. The wound healing was faster in α-lipoic acid-treated group than control group. At 7 and 14 days, the relative wound area and miR-29b expression level were significantly lower, while the contents of SOD and GSH were significantly higher in α-lipoic acid-treated group than control group (P < 0.05). In addition, miR-29b mimic group had significantly increased relative wound area and significantly decreased the contents of SOD and GSH when compared with NC group at 7 and 14 days (P < 0.05). Conclusionα-lipoic acid could inhibit oxidative stress and promote diabetic wound healing by suppressing expression of miR-29b in mice.
ObjectiveTo evaluate the effects of N-acetylcysteine (NAC) on lung tissue of Wistar rats, which were tracheally instilled fine particulate matter (PM2.5).MethodsForty-eight male Wistar rats were randomly divided into six groups: two control groups [they were blank group (C1), fake treatment group (C2) separately], four treatment groups [they were PM2.5 group (P), low-dose NAC group (L), medium-dose NAC group (M), high-dose NAC group (H) separately]. C1 received no treatments at all. C2 was instilled with sterile water (1 ml/kg) tracheally once a week for four times. P was instilled equivoluminal PM2.5 suspension (7.5 mg/kg) tracheally once a week for four times. The NAC groups received gavage (10 ml/kg) of different dosage of NAC (125, 250, 500 mg/kg) for six days. At the seventh day, the NAC groups were instilled PM2.5 suspension (7.5 mg/kg) tracheally. The procedures were repeated for three times in the NAC groups. Twenty-four hours later after four weeks or after the last instilling, all rats were sacrificed. Lung tissue was stained by hematoxylin-eosin (HE) staining, and histopathological changes of lung tissue were observed by optical microscope. The levels of C-reactive protein (CRP) as well as tumor necrosis factor-α (TNF-α) of serum, TNF-α of bronchoalveolar lavage fluid (BALF), TNF-α as well as interleukin-1β (IL-1β) of homogenates of lung tissue were detected by enzyme-linked immunosorbent assay. The activity of lactate dehydrogenase (LDH) as well as the levels of malondialhyde (MDA) of serum and BALF were detected by standard colorimetric method.ResultsHE staining showed that the normal structure of lung were destroyed in the groups dealed with PM2.5 and NAC could alleviate these changes. Higher dosage of NAC seemed to provide more powerful protections. Structure of the lung in C1 as well as C2 were nearly normal. The levels of CRP as well as TNF-α of serum, TNF-α of BALF, TNF-α as well as IL-1β of homogenates of lung tissue in the groups of P, L, M, H were higher than that in the groups of C1, C2 (all P<0.05). The levels of CRP as well as TNF-α of serum, TNF-α of BALF, TNF-α as well as IL-1β of homogenates of lung tissue in the groups of L, M, H which groups received NAC treatments were lower than that in P group. More, the groups seemed to have lower levels of CRP, TNF-α, IL-1β when higher dosage of NAC were given. The activity of LDH as well as the levels of MDA of serum, and BALF in the groups of P, L, M, H were higher than that in the groups of C1, C2 (all P<0.05). The activity of LDH as well as the levels of MDA of serum and BALF in the groups of L, M, H which groups received NAC treatments were lower than that in P group (all P<0.05). ConlusionTo some extent, NAC demonstrate antagonistic effects on oxidative stress and inflammatory injury on rats’ lung brought by PM2.5.
Objective To evaluate the expression of reactive oxygen species (ROS), glutataione (GSH), total superoxide dismutase (T-SOD), total antioxidation capacity (T-AOC), thioredoxin reductase (TrxR) under the intervention of hedysarum polysaccharides-1 (HPS-1) in A549 cells. Methods After treated by HPS-1 in different doses (50 mg/L, 100 mg/L, 200 mg/L, respectively), the viability of cell lines was detected by MTT method under microscope. The apoptosis of cell lines was detected by flow cytometry (FCM). The expressions of ROS, GSH, T-SOD, T-AOC, and TrxR in cell supernatant were measured by chemiluminescence method. Results Determined by MTT/FCM/ELISA, the results showed that different doses of HPS-1 could inhibit the proliferation and promote the apoptosis of A549 cells (allP<0.05). The expression levels of GSH, T-SOD, T-AOC, and TrxR were significantly decreased (allP<0.05) and the expression levels of ROS and MDA were significantly increased (allP<0.05) in a concentration-dependent manner in A549 cells treated with HPS-1, and these effects were significantly weakened in A549 cells with time extending (allP<0.05). Conclusion HPS-1 has a markedly effect on inhibiting cellular proliferation and inducing cellular apoptosis of lung adenocarcinoma A549 cells, which may be associated with the change of oxidation/antioxidant.
ObjectiveTo investigate the protective effect of mangiferin on acute spinal cord injury (SCI) in rats and its mechanism. MethodsNinety Sprague Dawley rats were randomly divided into 5 groups, 18 rats in each group. SCI was induced by using the Allen's method (60 g/cm) at T9 level in the rats of groups B, C, D, and E; laminectomy was performed at T8-10 in group A. The rats were injected intraperitoneally with saline in groups A and B, and with mangiferin in groups C (10 mg/kg), D (25 mg/kg), and E (50 mg/kg) every day for 30 days. The survival condition of rats was observed after operation; at 24, 48, and 72 hours after operation, the motor function of the hind limb was evaluated by the Basso, Beattie, Bresnahan (BBB) scores. The spinal cord edema was assessed by measuring the water content in spinal cord tissues at 72 hours. Meanwhile, malondialdehyde (MDA), catalase (CAT), superoxide dismutase (SOD), and glutathione peroxidase (GSH) were detected by ELISA; nuclear factor κB (NF-κB), tumor necrosis factor α (TNF-α), interleukin 1β (IL-1β), and IL-6 were measured via ELISA at the same time. Caspase-3 and Caspase-9 were also detected by ELISA after mangiferin treatment for 30 days. The expressions of Bax and Bcl-2 proteins were detected by Western blot. Pathological changes of the spinal cord was observed by HE staining. And Caspase-3 protein expression was detected by immunohistochemical staining. ResultsAll rats survived to the end of experiment. BBB scores of groups B, C, D, and E were significantly less than that of group A (P < 0.05), and it showed an increase trend from groups B to E (P < 0.05). The content of water of groups B, C, D, and E were significantly greater than that of group A (P < 0.05), and it showed a decrease trend from groups B to E (P < 0.05). ELISA showed that the activities of MDA, NF-κB, TNF-α, IL-1β, IL-6, Caspase-3, and Caspase-9 in groups B, C, D, and E were significantly greater than that in group A (P < 0.05), and they showed decrease trends from groups B to E (P < 0.05). Meanwhile, the activities of CAT, SOD, and GSH in groups B, C, D, and E were significantly less than that in group A (P < 0.05), and they showed increase trends from groups B to E (P < 0.05). Western blot showed that the relative expression of Bax protein in groups B, C, D, and E were significantly greater than that in group A (P < 0.05), and it showed a decrease trend from groups B to E (P < 0.05). Meanwhile, the relative expression of Bcl-2 protein in groups B, C, D, and E were significantly less than that in group A (P < 0.05), and it showed an increase trend from groups B to E (P < 0.05). Histological observation showed that the pathological changes in group B were accord with that in SCI, and the degree of necrosis in groups C, D, and E were significantly improved when compared with that in group B, and the effect was better in group E than group D, and group D than group C. Immunohistochemical staining showed that the absorbance (A) value of Caspase-3 in groups B, C, D, and E were significantly greater than that in group A (P < 0.05), and it showed a decrease trend from groups B to E (P < 0.05). ConclusionMangiferin has neuroprotective effects on acute SCI in rats by alleviating edema of spinal cord, inhibiting oxidative stress and inflammation response, and regulating the Bcl-2 and Bax pathway.
Objective To observe and preliminarily explore the effect of mogroside on oxidative stress of retinal pigment epitheliaum (RPE) cells induced by hydrogen peroxide (H2O2) and its possible mechanism. MethodsA experimental study. The RPE cells were divided into control group, H2O2 group, silent information regulator of transcription 1 (SIRT1) inhibitor EX527 group (EX527 group), mogroside group, mogroside+EX527 group. Methyl thiazolete trazolium method was used to detect cell survival rate. Flow cytometry was used to detect cell apoptosis rate. 2',7'-dichlorodihydrofluorescein diacetate fluorescent probe method, xanthine method and enzyme-linked immunosorbent assay method were used to detect the level of reactive oxygen species (ROS), superoxide dismutase (SOD) activity and malondialdehyde (MDA) content in cells respectively. Real-time quantitative polymerase chain reaction and Western blot were used to detect relative expressions of SIRT1, nuclear factor erythroid-2-related actor 2 (Nrf2), heme oxygenase-1 (HO-1) mRNA and protein in cells. One-way ANOVA was used for comparison among groups. The pairwise comparison between groups was tested by the least significant difference t test. Results Compared with the control group, the H2O2 group cell survival rate decreased, the apoptosis rate increased, the ROS level in the cells increased, the SOD activity decreased, the MDA content increased, and the relative expression of SIRT1, Nrf2, HO-1 mRNA and protein decreased (P＜0.05). Compared with H2O2 group, the cell survival rate decreased, apoptosis rate increased, the cell ROS level increased, SOD activity decreased, MDA content increased, SIRT1, Nrf2, HO-1 mRNA and protein expression decreased in EX527 group (P＜0.05); the cell survival rate increased, apoptosis rate decreased, ROS level decreased, SOD activity increased, MDA content decreased, and the relative expression of SIRT1, Nrf2, HO-1 mRNA and protein increased in mogroside group (P＜0.05). Compared with the mogrosides group, the cell survival rate decreased, the apoptosis rate increased, the level of ROS increased, SOD activity decreased, MDA content increased, SIRT1, Nrf2, HO-1 mRNA and protein decreased in mogrosides+EX527 group (P＜0.05). ConclusionsMogrosides can alleviate the oxidative stress response of visual RPE cells induced by H2O2, promote cell proliferation, and reduce cell apoptosis. Mogrosides may exert antioxidant effects by activating the SIRT1/Nrf2 signaling pathway.
Objective To explore repair role of allogeneic bone marrow mesenchymal stem cells （BM-MSCs） transplantation on treating hepatic ischemia reperfusion injury （HIRI） in rats. Methods Ten rats were executed to get BM-MSCs， then BM-MSCs were cultured in vitro and dyed by 4，6-diamidino-2-phenylindole （DAPI）. Models of 70% hepatic ischemia reperfusion injury were eatablished. Thirty two rats were randomly divided into sham operation group （Sham group）， ischemia reperfusion group （I/R group）， Vitamin C group （VC group）， and BM-MSCs group. Serum samples were analyzed for ALT and AST， and hepatic tissue were for superoxide dismutase （SOD） and malondialdehyde （MDA）. Liver sections were stain with hematoxylin and eosin （HE） for histological analysis， TUNEL staining was applied to detect hepatic apoptosis. Serum and tissues were both collected at 24 h after reperfusion. Results The isolated BM-MSCs maintained vigorous growth in vitro. Specific markers for MSCs antigens CD29 and CD44 were detected by flow cytometry， but antigens CD34 and CD45 were not be detected. Models of HIRI were stable， and BM-MSCs were detected around the periportal area by DAPI staining. Compared with I/R group， levels of ALT， AST， MDA， and AI in the VC group and BM-MSCs group decreased at 24 h after reperfusion （P＜0.05）， meanwhile SOD level increased （P＜0.05）. Compared with VC group， levels of ALT， AST， MDA， and AI in the BM-MSC group decreased at 24 h after reperfusion （P＜0.05）， meanwhile SOD level increased （P＜0.05）. Conclusion BM-MSCs could protect HIRI by alleviating oxidative stress and inhibiting cellular apoptosis.
Objective To evaluate the effects of N-acetylcysteine ( NAC) on bleomycin-induced lung fibrosis in mice and to investigate the therapeutic mechanisms of NAC on lung fibrosis. Methods Forty-five KM female mice were randomly divided into 3 groups. The mice in the control group were administered with saline aerosol intratracheally. The mice in the fibrosis group were administered with bleomycin ( 3 mg/kg) dissolved in normal saline aerosol intratracheally. The mice in the NAC group were gastric perfused with NAC at a dose of 400 mg · kg- 1 · d - 1 after administering bleomycin aerosol intratracheally. All animals were sacrificed 28 days after the treatments. The left lung was fixed in 10% neutral formalin, then stained with hematoxylin eosin and Masson’s trichrome respectively for the pathological examination. The right lung was sampled and the content of hydroxyproline ( HYP) was assayed by alkaline hydrolysis method. The serum was collected and the concentrations of malondialdehyde ( MDA) and totalantioxidant capacity ( T-AOC) were measured by colorimetric method. The RNA and total tissue protein were extracted for the examination of NOX1 /2/4 by RT-PCR and Western blot respectively. Results NAC prevented lung fibrosis induced by bleomycin with significantly reducing lung collagen accumulation and the level of HYP in the NAC group ( P lt;0. 05) . The serum concentration of MDA were reduced and serum TAOC raised by treating NAC after intratracheal administration of bleomycin ( P lt;0. 05) . NOX1 /2/4 gene and protein expression were increased in the fibrosis group compared with the control group. NAC had no effect on the gene expression of NOX1/2 /4( P gt;0. 05) , but inhibitted the NOX4 protein expression in lung tissue significantly ( P lt; 0. 05) . Conclusion NAC inhibits the expression of NOX4 and prevents bleomycin-induced lung fibrosis in mice.
Objective To investigate the effects of adenosine 2A receptor (A2AR) activation on oxidative stress in small-forsize liver transplantation. Methods A rat orthotopic liver transplantation model was performed using 40% graft, 18 recipients were given intravenously saline (control group), CGS21680 (A2AR agonist, CGS21680 group) or ZM241385 (A2AR antagonist, CGS21680+ZM241385 group) randomly. Aspartate aminotransferase (AST), enzymatic antioxidants 〔superoxide dismutase (SOD); catalase (CAT); glutathione peroxidase (GSH-Px)〕, non-enzymatic antioxidants 〔ascorbic acid (AA); glutathione (GSH); α-tocopherol (TOC)〕 and lipid oxidant metabolites malondialdehyde (MDA) were measured and analyzed at 6 h after reperfusion. Results Compared with the control group and CGS21680+ZM241385 group, A2AR activation increased the activities of SOD and GSHPx (Plt;0.05), reduced the productions of AST and MDA (Plt;0.05), increased the levels of AA, GSH and TOC (Plt;0.05) in CGS21680 group. But there was no significant change in CAT activity (Pgt;0.05) among 3 groups. Conclusions A2AR activation improves the antioxidant enzyme activities, promotes the production of antioxidants, and slowes down the increase in MDA level, depresses of the increase in AST activity. A2AR activation suppresses oxidative damage and increases the antioxidant capacity which in turn minimizes their harmful effects of ischemia-reperfusion in small-for-size liver transplantation.
Objective To investigate the implication of oxidation protein product ( advanced oxidation protein product, AOPP) , an index of oxidative stress in obstructive sleep apnea-hypopnea syndrome ( OSAHS) . Methods 47 patients with OSAHS and 48 normal controls were enrolled. The concentration of AOPP was measured by spextrophotometry after ameliorated, while superoxide ( SOD) , malonaldehyde ( MDA) , glutathione peroxidase ( GSH-PX) in morning blood samples were detected by Xanthine oxidase test. Results ( 1) Plasma AOPP and MDA were significantly elevated in OSAHS compared with those in control group ( both P lt;0. 01) . Plasma SOD and GSH-PX were significantly lower in OSAHS compared with those in control group ( both P lt;0. 01) . There were significant differences in the plasma AOPP, MDA, SODand GSH-PX among different severity of OSAHS ( all P lt; 0. 01) . Plasma AOPP and MDA were increased and SOD and GSH-PX were gradually decreased with the progression of OSAHS. ( 2) Plasma AOPP correlated well with MDA, SOD and GSH-PX, moreover, AOPP was positively correlated with apnea hyponea index or lowest oxygen saturation. Conclusion AOPP is an alternative index reflecting both oxidative streess and tissue injury in patients with OSAHS.
Febrile seizures (FS) are one of the most common neurological disorders in pediatrics, commonly seen in children from three months to five years of age. Most children with FS have a good prognosis, but some febrile convulsions progress to refractory epilepsy (RE). Epilepsy is a common chronic neurological disorder , and refractory epilepsy accounts for approximately one-third of epilepsies. The etiology of refractory epilepsy is currently complex and diverse, and its mechanisms are not fully understood. There are many pathophysiological changes that occur after febrile convulsions, such as inflammatory responses, changes in the blood-brain barrier, and oxidative stress, which can subsequently potentially lead to refractory epilepsy, and inflammation is always in tandem with all physiological changes as the main response. This article focuses on the pathogenesis of refractory epilepsy resulting from post-febrile convulsions.