Age-related macular degeneration (AMD) is one of the leading irreversible causes of blindness in China. The pathogenesis of AMD is not fully understood at present. Under various stress conditions, cellular senescence is activated, characterized by telomere shortening, mitochondrial dysfunction, DNA damage, and the release of various senescence-related secretory phenotype factors. Senescence is implicated in the pathogenesis of AMD through multiple pathways, contributing to chronic inflammation and the onset and progression of AMD. Mechanisms such as oxidative stress, lipofuscin, β amyloid protein and the membrane attack complex have become hotspots of study in the pathogenesis of AMD. The cyclic guanosine phosphate - adenosine synthase - interferon stimulating factor synthase-stimulator of interferon gene pathway has emerged as a critical signaling pathway in the early development of AMD, providing direction for further research on AMD. Currently, senolytics, selective agents targeting the induction of senescent cell apoptosis, show significant potential in the treatment of AMD. The integration of new technologies with cellular senescence may offer a novel approach to AMD treatment, and intervening in the AMD treatment through anti-cellular senescence pathways holds promising prospects.
Objective Bone marrow mesenchymal stem cells (BMSCs) transplantation can potentially regenerate the degenerated intervertebral disc, with the underlying regenerating mechanism remaining largely unknown. To investigate the potential of human BMSCs protecting nucleus pulposus cells (NPCs) from oxidative stress-induced apoptosis in a coculturesystem, and to illustrate the possible mechanisms of BMSCs transplantation for intervertebral disc regeneration. Methods BMSCs collected by density gradient centrifugation in Percoll solution were cultured and sub-cultured till passage 3, and the surface molecules of CD34, CD45, and CD13 were identified. NPCs were isolated by collagenase digestion and the chondrocyte l ike phenotype was confirmed by morphologic observation after HE staining, inverted phase contrast microscope, proteoglycan, and collagen type II expression after toluidine blue and immunocytochemistry staining. The 3rd passage BMSCs and the 1st passage NPCs were divided into four groups: group A, NPCs (1 × 106 cells) were cultured alone without apoptosis inducing (negative control); group B, NPCs (1 × 106 cells) were co-cultured with BMSCs (1 × 106 cells) with apoptosis inducing; group C, NPCs (1 × 106 cells) were co-cultured with BMSCs (3 × 105 cells) with apoptosis inducing; group D, NPCs (1 × 106 cells) were cultured alone with apoptosis inducing (positive control). After 3 or 7 days of culture or co-culture, the NPCs in groups B, C, and D were exposed to 0.1 mmol hydrogen peroxide for 20 minutes to induce apoptosis. With DAPI staining cellular nucleus, Annexin-V/propidium iodide staining cellular membrane for flow cytometry analysis, the apoptosis of NPCs in each group was studied both qual itatively and quantitatively. Besides, the changes in Bax/Bcl-2 gene transcription and Caspase-3 protein content, were analyzed with semi-quantitative RT-PCR and Western blot. Results BMSCs were successfully isolated and CD34-, CD45-, and CD13+ were demonstrated; after isolated from degenerated intervertebral discs and sub-cultured, the spindle-shaped 1st passage NPCs maintained chondrocyte phenotype with the constructive expressions of proteoglycan and collagen type II in cytoplasm. DAPI staining showed the nucleus shrinkage of apoptosis NPCs. Co-cultured with BMSCs for 3 days and 7 days, the apoptosis rates of NPCs in groups B (29.26% ± 8.90% and 18.03% ± 2.25%) and C (37.10% ± 3.28% and 13.93% ± 1.25%) were lower than that in group D (54.90% ± 5.97% and 26.97% ± 3.10%), but higher than that of groupA (15.67% ± 1.74% and 8.87% ± 0.15%); all showing significant differences (P lt; 0.05). Besides, semi-quantitative RT-PCR showed Bcl-2 gene transcription up-regulated (P lt; 0.05) and no significant change of Bax (P gt; 0.05); Western blot result showed that the Caspase-3 protein expression of groups B and C was lower than that of group D, and was higher than that of group A; all showing significant differences (P lt; 0.05). Conclusion In a co-culture system without direct cellular interactions, the oxidative stress-induced apoptosis of human NPCs was amel iorated by BMSCs. The enhanced anti-apoptosis abil ity of NPCs preconditioned by co-culturing with BMSCs might come from the decreased Bax/Bcl-2 gene transcription ratio.
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.
Objective To evaluate the effects of oxidative stress in the airway inflammation and remodeling of high-fat diet induced obese mice with asthma. Methods Sixty female C57 /6J mice were randomly divided into four groups, ie. an asthma group, an obese group, an obese asthma group, and a control group. The mice in the asthma group were sensitized and challenged with ovalbumin ( OVA) and fed with normal diets. The mice in the obese group were fed with high-fat diets. The mice in the obese asthma group were sensitized and challenged as the asthma group, and fed as the obese group. The mice in the control group were sensitized and challenged with normal saline and fed with normal diets. After 12 weeks, bronchoalveolar lavage fluid ( BALF) were collected for total and differential cell count. IL-6 and 8-iso-prostaglandin F2α ( 8-iso-PGF2α) in lung tissue homognate were detected by ELISA. The pathological changes were observed under light microscope by HE staining. Meanwhile the remodeling indices including total bronchial wall area ( WAt) , smooth muscle area ( WAm) , and bronchial basement membrane perimeter ( Pbm) were measured. Results In comparison with the obese group and the asthma group, the leukocytes and eosinophils in BALF, WAt/ Pbm, and IL-6 in lung tissue increased significantly in the obese asthma group ( P lt; 0. 05) . 8-iso-PGF2αin lung tissue increased in sequence of the control group, the obese group, the asthma group, and the obese asthma group significantly. Pearson correlation analysis showed that leukocyte in BALF, WAt/ Pbm, and IL-6 were in positive correlation with 8-iso-PGF2α( r =0. 828, 0. 863, 0. 891, respectively, P lt;0. 01) . Conclusion Oxidative stress is involved in the airway inflammation and remodeling of obese asthma mice with high-fat diets.
ObjectiveTo systematically review the association of micronucleus and polycystic ovary syndrome (PCOS).MethodsPubMed, OVID, Elsevier, CNKI, WanFang Data and CBM databases were electronically searched to collect case-control studies on the association of micronucleus and PCOS. Two reviewers independently screened literature, extracted data and assessed the risk of bias of included studies, then, meta-analysis was performed by using RevMan 5.3 software.ResultsA total of 5 case-control studies were included, in which 170 patients were in the case group and 148 in the control group. The results of meta-analysis showed: there were significant differences between the two groups for micronucleus frequency (MD=2.02%, 95%CI 1.63% to 2.41%, P<0.000 01) in peripheral blood lymphocytes and micro nucleated cells frequency (MD=2.43%, 95%CI 0.10% to 4.76%, P=0.04) in oral epithelial cells.ConclusionThe current evidence shows that micronucleus is associated with PCOS. Due to limited quality and quantity of the included studies, more high quality studies are needed to verify above conclusion.
ObjectiveTo study the effect of rotenone on rat substantia nigra dopamine (DA) in the nervous system and oxidative stress parameters (malondialdehyde and glutathione), the influence of rotenone on DA neurons toxic effect and its pathogenesis.
MethodsThis study applied back subcutaneous injection of rotenone in rats [1.0 mg/(kg·d)], and used immunocytochemistry technique to detect changes in the expression of tyrosine kinase (TH) in 10 rats of the control group and 10 rats of the experimental group. Spectrophotometry was used to detect the change of oxidative stress parameters in rats (malondialdehyde and glutathione).
ResultsDA neurons in rats had various degrees of damage. The TH immune response strength of rats in the substantia nigra and striatum decreased significantly. The number of immune response nigra TH positive neurons was significantly less in the experimental group than in the control group (P< 0.01). Spectrophotometer method was used to detect the midbrain nigra of glutathione, which was significantly less in the experimental group than in the control group (P<0.01). Malondialdehyde in the experimental group was significantly higher (P<0.01).
ConclusionRotenone has obvious neurotoxicity, and can lead to the damage of DA neurons and obvious oxidative stress injury in rats, which provides an experimental basis for the pathogenesis of Parkinson's disease, and at the same time provides new targets for the treatment.
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.
ObjectiveTo investigate the effect of diammonium glycyrrhizinate (DG) plus bone marrow mesenchymal stem cells (MSCs) transplantation in the treatment of acute exacerbation of pulmonary fibrosis induced by bleomycin (BLM) in rats.MethodsMSCs were isolated from male Wistar rats and cultured in vitro. Twenty-four female Wistar rats were randomly divided into 4 groups. The NC group was intratracheally injected with normal saline; the BLM group, the MSC group and the DGMSC group were intratracheally injected with BLM for 7 days; then the MSC group was injected with 0.5 mL of MSCs solution (2.5×106 cells) into the tail vein; the DGMSC group was intraperitoneally injected with DG for 21 days in a dose of 150 mg·kg–1·d–1 on the base of the MSCs injection. The rats were sacrificed on the 28th day and the lung tissue was extracted. Pathological examination was performed to determine the degree of alveolitis and pulmonary fibrosis. Immunofluorescence was used to detect the number and distribution of alveolar type Ⅱ epithelial cells. Alkali hydrolysis method was used to determine the content of hydroxyproline (HYP) in lung tissue; thiobarbituric acid method was used to measure the content of malondialdehyde (MDA) in lung tissue; colorimetric method was used to determine the superoxide dismutase activity (SOD) and total antioxidant capacity (T-AOC); enzyme linked immunosorbent assay was used to detect the expression levels of tumor necrosis factor-α (TNF-α ) and transforming growth factor-β1 (TGF-β1) in lung tissue homogenates.ResultsThe DG combined with MSCs injection can reduce the degree of alveolitis and pulmonary fibrosis in BLM model rats. The content of HYP and TGF-β1 in lung tissue homogenate of the DGMSC group were significantly lower than those in the MSC group (P<0.05). Meanwhile, DG combined with MSCs injection significantly increased the antioxidant capacity of the BLM model rats. MDA content decreased, SOD activity and T-AOC ability improved significantly in the DGMSC group compared with the MSC group (P<0.05). The alveolar type Ⅱ epithelial cells were significantly increased and the cell morphology was maintained in the DGMSC group compared with the MSC group.ConclusionsDG has a synergistic effect with MSCs in treatment of acute exacerbation of pulmonary fibrosis. The mechanism may be related to reducing inflammatory factors during pulmonary fibrosis, attenuating oxidative stress and promoting MSCs migration into lung tissue and transformation to alveolar type Ⅱ epithelial cells.
Abstract: Objective To determine the effects of oxidative stress reaction on intima hyperplasia after autologous vein grafting. Methods Seventy female SpragueDawley(SD) rats were randomly divided into a control group(n=10) and an experimental group (n=60). The experimental group was then divided into six time points of one day; one, two, four, and six weeks; and two months after surgery; with 10 rats for each time point. Autologous vein grafting models were established. At each time point the designated rats were anaesthetized, and the grafts were isolated and stained with HE. The same length of external jugular vein was cut from each rat in the control group. The neointima to tunica media area ratios (I/M) were measured with acomputerized digital image analysis system. Nuclear factorkappa B (NF-κB) and copper zinc superoxide dismutase (CuZnSOD) were detected byimmunohistochemistry. The concentration of malondialdehyde (MDA) in serum was analyzed by colorimetry. Results In the control group, expression levels of NF-κB and CuZnSOD were low. In the experimental group, expression of NF-κB increased after the operation and peaked two weeks later. The plateau was sustained for about one month, and then the level of expression declined gradually, reaching the baseline at the twomonth time point. The expression of CuZnSOD increased gradually after the operation and peaked one week later, then declined to the normal level after 2-3 weeks at the plateau. In the control group, the concentration of serum MDA was 4.966±1.346 nmol/ml. In the experimental -group, the-MDA concentration increased dramatically after the operation, then-declined from its highest level at the oneday time point (21.161±2.174 nmol/ml) to the normal level at two months (6.208±2.908 nmol/ml) after the operation (P＜0.05). In the control group, I/M was 0.2096±0.0253, while in the experimental group, it was higher one week after the operation (0.6806±0.0737) and peaked at four weeks (1.4527±0.0824), falling to 1.0353±00656 at six weeks and 0.9583±0.0516 attwo months (P＜0.05) for the experimental and control groups). Conclusion Endothelial cell injury initiates an oxidative stress reaction after autologous vein grafting and augments inflammation by activating NF-κB, thus playing an important role in inducing restenosis of the grafted vein.
