MDA Expression Research Articles

MicroRNA-204 Attenuates Oxidative Damage in Cardiac Stem Cell Through Regulation of Bone Marrow Stromal Cell (BMSC) Adipogenic and Osteogenic Differentiation

Exosomes (exo) derived from bone marrow mesenchymal stem cells (BMSCs) are known to promote cell growth through delivering multiple kinds of bioactive molecule including microRNAs (miR-NAs). This study aimed to explore the mechanism underlying miR-204 secreted by exo interacting oxidative damage of cardiac stem cell (CSCs). Exosomes were extracted from BMSCs (BMSC-exo) and characterized by immunofluorescence and electron microscope, while BMSC-exo were internalized by CSCs. ARS and ALP staining confirmed the mineralization of BMSCs and osteogenic and adipogenic differentiation of BMSCs. Then BMSCs were cultured in ordinary culture medium (OM) and normal medium. RT-qPCR identified miR-204 level in BMSCs disposed by OM was about five times as that of controls. miR-204 was up-regulated in the osteogenic differentiation of CSCs. Functional experiment revealed up-regulation of miR-204 inhibited the BMSC adipogenic differentiation with decreased ROS and MDA expression and elevated SOD level in the CSCs. Treatment with BMSC-exos or miR-204 up-regulation alleviated oxidative damage of CSCs. Collectively, miR-204 attenuates the oxidative damage of CSCs through regulating BMSC adipogenic and osteogenic differentiation.

Carvacrol exerts nephroprotective effect in rat model of diclofenac-induced renal injury through regulation of oxidative stress and suppression of inflammatory response

Diclofenac (DIC) is an NSAID that can cause toxic effects in animals and humans and carvacrol (CAR) is a monoterpene compound that displays effective pharmacological and biological actions. The purpose of this work was to assess the influences of CAR on DIC-induced renal injury and oxidative stress in male rats. The rats were segregated into four groups. Group 1, control group; Group 2 received DIC-only; Groups 3, received CAR-only and group 4 received DIC plus CAR. Changes in biochemical indexes, pathological changes, molecular biological indexes, and genes related to the inflammation of main organs were evaluated. The results of this work indicated that the amounts of the serum protein carbonyl, sGOT, sGPT, urea, creatinine, uric acid, nitrite content, MDA, serum TNF-α, and renal TNF-α gene expression were remarkably increased and the levels of the GPx, GSH, CAT, and SOD were significantly reduced in DIC-only treated animals compared to the control group. On the other hand, treatment with CAR after exposure to DIC led to significant improvements in abnormalities of DIC-induced renal injury and serum biochemical factors. The data approve that CAR diminished the deleterious effects of DIC exposure. In this regard, the findings of this study indicated that the administration of CAR could alleviate the noxious effects of DIC on the antioxidant defense system and renal tissue.

Therapeutic Effects of Hydroalcoholic Extracts from the Ancient Apple Mela Rosa dei Monti Sibillini in Transient Global Ischemia in Rats.

The Mela Rosa dei Monti Sibillini is an ancient apple variety cultivated by Romans in the foothills of the Sibillini Mountains, central Italy, showing potential as a source of nutraceuticals. The purpose of this study was to evaluate the protective effects of the hydroalcoholic extracts from the peel (APE) and pulp (APP) of this fruit in an animal model of transient global ischemia. Chemical constituents were analyzed by liquid chromatography–mass spectrometry (LC-DAD-MSn) indicating several polyphenols such as B-type procyanidins, quercetin derivatives and hydroxycinnamic acids as the main bioactive components. Acute pre-treatment of extracts (30 mg/kg, i.p.) significantly decreased the brain levels of the pro-inflammatory cytokines IL-1β (p < 0.01) and TNF-α (p < 0.001 and p < 0.01 for APE and APP, respectively), the expression of caspase-3 (p < 0.01, For APE) and MDA (p < 0.05), a lipid peroxidation biomarker in rats. Both extracts restricted the pathological changes of the brain induced by ischemic stroke in hematoxylin and eosin assay. Moreover, they improved the scores of behavioral tests in grid-walking and modified neurological severity scores (mNSS) tests. In conclusion, these results proved this ancient Italian apple is a source of nutraceuticals able to protect/prevent damage from brain ischemia.

Correlation Between Oxidant and Antioxidant Status (MDA and SOD Expression) and the Improvement of Outer Hair Cells Function Due to Curcumin Administration in Noise-Exposed Rattus Norvegicus Cochlea Assessed by DPOAE Examination

Correlation Between Oxidant and Antioxidant Status (MDA and SOD Expression) and the Improvement of Outer Hair Cells Function Due to Curcumin Administration in Noise-Exposed Rattus Norvegicus Cochlea Assessed by DPOAE Examination

Optimal intervention time of ADSCs for hepatic ischemia-reperfusion combined with partial resection injury in rats

AimsHepatic ischemia reperfusion injury (HIRI) is a complication of liver surgery and liver transplantation. Adipose-derived stem cells (ADSCs) can inhibit oxidative stress and inflammation through a paracrine effect. This study aimed to determine the optimal time window of ADSCs transplantation to restore liver function after HIRI. Main methodsA rat model of hepatic ischemia reperfusion combined with partial hepatectomy (HIR/PH) was established. The animals were injected intravenously with 2 × 106 rat ADSCs 2 h before, immediately after, or 6 h after surgery. Liver tissues and blood samples were collected for routine histological and biochemical assays. The molecular changes were analyzed by qRT-PCR and western blotting. Key findingsADSCs significantly improved liver tissue structure and decreased the levels of AST, ALT and ALP, which was indicative of functional recovery. In addition, transplantation of ADSCs immediately after operation decreased the levels of inflammation-related cytokines such as TNF-α, IL-1β and IL-6, and significantly increased the activity of antioxidant enzymes. At the same time, the expression of MDA was decreased. Mechanistically, ADSCs activated the Keap1/Nrf2 pathway in the injured liver. Transplantation of ADSCs pre- and 6 h post-operation did not significantly affect some indices such as mRNA and protein expression of HO-1, and protein expression of NQO1. SignificanceTransplanting ADSCs immediately after surgery accelerated tissue repair and functional recovery of the liver by activating the Keap1/Nrf2 pathway, which inhibited hepatic inflammation and oxidative stress, and restored the hepatic microenvironment.

Minocycline attenuates oxidative and inflammatory injury in a intestinal perforation induced septic lung injury model via down-regulating lncRNA MALAT1 expression

BackgroundOxidative stress and inflammatory responses play an important role in acute lung injury (ALI). Although minocycline (MINO) has anti-inflammatory effects and is a promising candidate in treating inflammatory diseases, the effect of MINO on ALI during sepsis is still unclear. MethodsIn the present study, a mouse model with intestinal perforation was established. C57BL/6 mice received cecal ligation and puncture (CLP) to induce sepsis-associated ALI. MINO was used to treat the mice via intraperitoneal injection at different doses (negative control, 20 mg/kg, 50 mg/kg and 100 mg/kg, respectively) 24 h after CLP. The severity of lung injury was evaluated by pathological examination, and lung wet / dry weight ratio was calculated to evaluate the severity of pulmonary edema. The changes of TNF-α, IL-1β, IL-6, PGE2, MDA, NF-κB, Nrf2, Keap1 and lncRNA MALAT1 levels in lung tissues of the mice were detected with ELISA, chemical colorimetry, Western blot or qRT-PCR. ResultsMINO ameliorated the lung edema and lung injury of the mice induced by CLP in a dose-dependent manner. MINO administration could significantly down-regulate expressions of TNF-α, IL-6, IL-1β, PGE2 and MDA in lung tissues of the mice. Mechanistically, MINO exerted the effects of anti-inflammation and anti-oxidative stress through down-regulating the expression of MALAT1 and regulating Nrf2/Keap1 and NF-κB signaling pathways. ConclusionMINO represses oxidative stress and inflammatory response during sepsis-induced ALI via down-regulating MALAT1 expression, and it has the potential to treat septic ALI.

Ferroptosis is involved in the anti-tumor effect of lycorine in renal cell carcinoma cells.

Renal cell carcinoma (RCC) is a most common malignant tumor in the genitourinary system. Studies have shown that Lycorine has promising anticancer activities with minor side effects. However, the effect of lycorine on the proliferation of RCC cells and its underlying anti-tumor mechanism have not yet been fully elucidated. The human renal cancer cell lines 786-O, A498 and Caki-1 were cultured and treated with different concentrations of lycorine or ferrostatin-1, a ferroptosis inhibitor. Cell viability and colony formation assays were used to measure cell proliferation. The 5-, 12- and 15-HETE hydroxyeicosatetraenoic acid (HETE) and MDA levels, as well as the reduced to oxidized glutathione (GHS/GSSG) ratio, were analyzed. Western blot analysis was used to detect the expression of glutathione peroxidase 4 (GPX4) and acyl-CoA synthetase long chain family member 4 (ACSL4), which are key markers of ferroptosis. Transmission electron microscopy was used to observe the morphological features associated with ferroptosis. Lycorine was found to inhibit the proliferation of RCC cells. After lycorine treatment, the expression levels of GPX4 in RCC cells decreased, whereas those of ACSL4 increased. Lycorine induced the expression of 5-HETE, 12-HETE, 15-HETE and MDA in RCC cells, and reduced the GSH/GSSG ratio. In addition, ferrostatin-1 could prevent lycorine-induced ferroptosis in RCC cells.

Puerarin Inhibits Ferroptosis and Inflammation of Lung Injury Caused by Sepsis in LPS Induced Lung Epithelial Cells

Background: Ferroptosis is a new type of programmed cell death, which plays an important role in lung injury caused by sepsis. Studies have reported that Puerarin (Pue) can treat lung injury caused by sepsis in children, but whether it plays a role by regulating iron death has not been reported.Methods: LPS induced human alveolar epithelial cell A549 to form a model of lung injury caused by sepsis. MTT detected the effect of Pue on A549 cell viability and the effect of Pue on LPS-induced A549 cell viability. The effects of Pue on LPS-induced inflammatory cytokines TNF-α, IL-8, IL-1β in A549 cells were determined by ELISA assay. The expression level of MDA was detected by TBARS colorimetric quantitative detection kit. GSH kit was used to detect the expression of GSH in cells. The iron kit detected the total iron level and the expression level of ferric divalent ions in the cells. DCFH-DA fluorescent probe was used to detect ROS levels. Western blot was used to detect the expression of ferroptosis-related proteins in cells.Results: Pue alleviated LPS-induced injury and inflammatory response in A549 cells, and Pue reduced the expression of ROS, MDA and GSH in LPS-induced A549 cells. In addition, Pue reduced total iron levels and ferrous ion levels in LPS-induced A549 cells, and decreased the expression of iron ferroptosis-related proteins.Conclusion: Puerarin inhibited ferroptosis and inflammation of lung injury caused by sepsis in children in LPS induced lung epithelial cells.

Abstract 2005: ZDHHC7 contributes to ferroptosis resistance through YAP localization in ovarian clear cell carcinoma

Abstract [Objective] Ovarian clear cell carcinoma (CCC) shows unique clinical features including an association with benign endometriosis which contains iron-rich fluid, and poor prognosis because of resistance to chemotherapy. Therefore, elucidating the basic mechanism as well as new treatment strategy of ovarian CCC is urgently needed. Recently, ferroptosis, which is characterized by the iron-dependent accumulation of lipid ROS, was reported to affect the malignant behaviors. Also, some reports showed ferroptosis regulation by Hippo signaling. ZDHHC7 is reported as the molecule regulating yes-associated protein (YAP), a key molecule of Hippo signature and is one of the ovarian CCC gene signature which we previously identified. We explored the impact of ferroptosis in ovarian CCC by investigating YAP localization.[Methods] We performed immunohistochemistry (IHC) of malondialdehyde (MDA; a marker of lipid reactive oxygen species, ROS) and 8-Oxo-2'-deoxyguanosine (8-OHdG, a marker of oxidative stress DNA damage) to analyze the status of lipid ROS with ovarian cancer patients. We also performed IHC of YAP and ZDHHC7 to evaluate them in clinical status. We calculated IC50 values of erastin (ferroptosis inducer) with seventeen ovarian cancer cell lines. Functional annotation clustering of microarray expression data was performed using Metascape. ZDHHC7 knockdown and YAP-5SA mutant, or YAP-activated form of ovarian CCC cell lines were generated and applied for in vitro assays.[Results] IHC analysis indicated higher MDA expression (p=0.0354) and 8-OHdG (p&amp;lt;0.0001) in CCC compared to high-grade serous carcinoma (HGSC), suggesting higher lipid ROS as well as oxidative stress status in ovarian CCC. IC50 values of erastin were significantly higher in CCC than in other types of ovarian cancer (p&amp;lt;0.01). Categorical analysis revealed that genes related to Hippo signaling were enriched in erastin-resistant cell lines. IHC analyses indicated low nuclear expression of YAP and high expression of ZDHHC7 in CCC samples (p=0.0010). The cases with low expression of nuclear YAP showed significantly poorer prognosis compared to high expression cases (p=0.0075) in ovarian CCC. YAP-5SA mutant (YAP-active-mutant) CCC cells showed higher sensitivity to erastin and ML210 (ferroptosis inducer) than control in ovarian CCC cell lines. ZDHHC7 knockdown of CCC cells induced nuclear localization of YAP and showed decreased IC50 values of erastin and ML210 compared to control cells.[Conclusion] Although lipid ROS is accumulated in ovarian CCC, ovarian CCC showed ferroptosis resistance through YAP localization regulated by ZDHHC7. Citation Format: Yoko Furutake, Ken Yamaguchi, Masayo Ukita, Mana Taki, Koji Yamanoi, Junzo Hamanishi, Takashi Kobayashi, Masaki Mandai. ZDHHC7 contributes to ferroptosis resistance through YAP localization in ovarian clear cell carcinoma [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2021; 2021 Apr 10-15 and May 17-21. Philadelphia (PA): AACR; Cancer Res 2021;81(13_Suppl):Abstract nr 2005.

The effect of aqueous extract of Prunus dulcis on tibial bone healing in the rabbit

BackgroundBone fractures are medical emergencies that require prompt intervention to help return bone to its normal function. Various methods and treatments have been utilized to increase the speed and efficiency of bone repair. This study aimed to investigate the treatment effects of Prunus dulcis aqueous extract on tibial bone healing in rabbits.MethodsAll animals were distributed in five groups with six rats in each group, including the sham group, the control group in which tibial lesion was made and received distilled water, treatment groups with 150 mg kg−1, 300 mg kg−1 doses of Prunus dulcis extract, and osteocare treated group. Biochemical blood factors including calcium, phosphorus, and alkaline phosphatase (on days 0, 10, 30, and 50), biomarkers of oxidative stress such as GPx, CAT, and MDA (on days 10 and 30), radiological evaluation, histopathological parameters, and osteocalcin immunohistochemical expression were assessed.ResultsThe data showed calcium levels in the treatment groups increased significantly from day 10 to day 50, respectively, and blood phosphorus levels decreased from day 10 to day 50 in the treatment groups. Alkaline phosphatase initially increased and then decreased in treatment groups. In the treatment groups, GPx and CAT levels significantly increased, and the serum amount of MDA reduced. The best antioxidant results were related to the extract-treated group with a higher dose. Radiographic score was significantly higher in the treatment groups than the control group on day 30. Based on the pathological findings, the healing occurred faster in the extract-treated group with a higher dose. Osteocalcin expression was significantly higher in the control group than that in the treatment groups.ConclusionsTreatment with Prunus dulcis extract with a dosage of 300 mg/kg accelerated tibial bone healing in rabbits.Graphical abstract

Metabolomics of Spleen-Yang deficiency syndrome and the therapeutic effect of Fuzi Lizhong pill on regulating endogenous metabolism

Ethnopharmacological relevanceSpleen-Yang deficiency (SYD) is one of the primary causes of many digestive diseases, such as ulcerative colitis (UC), and irritable bowel syndrome (IBS), but its endogenous metabolic characteristics are still unclear. Fuzi Lizhong pill (FLZP) is well-known for its powerful capacity for treating SYD; however, its mechanisms require further study. Aim of the studyHerein, our present study aimed to investigate the essence of SYD from the perspective of metabolomics, and tried to reveal the anti-SYD action mechanisms of FLZP. Materials and methodsFirstly, the compound factor modeling method with the principle of “indiscipline in diet + excessive fatigue + intragastric administration of Senna water extracts” was used to establish Sprague Dawley (SD) rats as SYD model. Then, the visceral index, motilin (MTL), malonaldehyde (MDA), Interleukin 1 alpha (IL-1α), and Interleukin 6 (IL-6) levels were used to verify the anti-SYD effect of FLZP. In addition, serum samples were analyzed by UPLC-QE/MS metabolomics technique. Finally, the metabolic pathways associated with specific biomarkers were analyzed to research the possible mechanism underlying the action of FLZP. ResultsThe expression of MTL, MDA, IL-1α, and IL-6 were regulated by FLZP, which suggested that it has relieved diarrhea and gastrointestinal motility disorder caused by SYD and had an anti-peroxidation, anti-inflammatory, and immune regulation effect. A total of 75 metabolites were found to be the potential biomarkers of SYD. Moreover, FLZP regulates 21 metabolites and 10 vital pathways including the tricarboxylic acid (TCA) cycle, sphingolipid metabolism, and histidine metabolism. ConclusionSYD primarily causes disorders of amino acid metabolism, lipid metabolism, carbohydrate metabolism, metabolism of cofactors and vitamins, nucleotide metabolism, and translation. In addition, FLZP regulated carbohydrate, lipid, and amino acid metabolisms, gastrointestinal motility, digestive juice secretion, immune regulation, as well as antioxidant effects. Hence, FLZP had a good therapeutic effect on treatment of SYD. It might be a promising therapeutic agent for the treatment of SYD-related diseases.

Mechanism of Yifei Decoction Combined with MitoQ on Inhibition of TGFβ1/NOX4 and PDGF/ROCK Signal Pathway in Idiopathic Pulmonary Fibrosis.

Background Rho-related coiled helix forming protein kinase (Rho-ROCK) and another important fibrogenic factor-PDGF play a critical role in collagen deposition in rat lung tissue. Yifei decoction (YFT), a Chinese herbal decoction, has been used to treat idiopathic pulmonary fibrosis (IPF) in clinical practice and has produced positive outcomes; however, convincing evidence is currently lacking. The present study aimed to investigate the effects of YFT combined with MitoQ in rats with IPF and to explore the underlying mechanism. Methods Rat IPF model was established by endotracheal injection of 5 mg/kg BleomycinA5 into the specific pathogen-free SD rats. MitoQ (6.5 μmol/kg once daily), YFT (10 ml/kg once daily), and MitoQ + YFT (6.5 μmol/kg + 10 ml/kg once daily) were used to treat the rat model for 4 weeks, respectively. The normal rats without IPF were used as the controls. After 4 weeks of drug treatment, lung histopathology was assessed. Immunohistochemistry was used to detect the expression of fibronectin and collagen IV in lung tissue. The expression of IL-6, IL-1β, TNF-α, GSH-Px, SOD, MDA, and hydroxyproline was determined by enzyme-linked immunosorbent assay. The expressions of TGFβ1, NOX4, PDGFR-β, and ROCK1 were determined using real-time quantitative PCR and Western blot. Results After 4 weeks of drug treatment, comparison of the MitoQ + YFT group with the IPF group showed that lung injury scores, W/D, lung tissue hydroxyproline, fibronectin, collagen IV content, and IL-6, IL-1β, TNF-α, and MDA levels were significantly lower (P < 0.05), as well as the expression of TGFβ1, NOX4, PDGFR-β, and ROCK1, but the activity of GSH-Px and SOD was higher (P < 0.05). Conclusion MitoQ combined with YFT can improve lung injury in rats with pulmonary fibrosis by reducing the secretion of proinflammatory cytokines and inhibiting TGFβ1/NOX4 and PDGF/ROCK signaling pathways. It may provide a new method for the treatment of pulmonary fibrosis.

MiR-485 inhibits histone deacetylase HDAC5, HIF1α and PFKFB3 expression to alleviate epilepsy in cellular and rodent models.

We investigated the role of microRNA (miR)-485 and its downstream signaling molecules on mediating epilepsy in cellular and rat models. We established a cellular epilepsy model by exposing hippocampal neurons to magnesium and a rat model by treating ICR mice with lithium chloride (127 mg/kg) and pilocarpine (30 mg/kg). We confirmed that miR-485 could bind and inhibit histone deacetylase 5 (HDAC5) and then measured expression of miR-485 and in mice and cells. Cells were transfected with overexpression or knockdown of miR-485, HDAC5, hypoxia-inducible factor-1alpha (HIF1α), or 6-phosphofructo-2-kinase/fructose-2,6-biphosphatase 3 enzyme (PFKFB3) to verify their roles in apoptosis, oxidative stress, and inflammation in epileptic hippocampal neurons. Binding relationship between miR-485, HDAC5, HIF1α, and PFKFB3 was verified. Oxidative stress and inflammation marker levels in epilepsy model mice were assessed. miR-485 was downregulated and HDAC5 was upregulated in cell and animal model of epilepsy. Seizure, neuronal apoptosis, oxidative stress (increased SOD and GSH-Px expression and decreased MDA and 8-OHdG expression) and inflammation (reduced IL-1β, TNF-α, and IL-6 expression) were reduced by miR-485 in epileptic cells. HIF1α and PFKFB3 expression was reduced by HDAC5 knockdown in cells, which was recapitulated in vivo. Thus, miR-485 alleviates neuronal damage and epilepsy by inhibiting HDAC5, HIF1α, and PFKFB3.

Allium jesdianum hydro alcoholic extract ameliorates diabetic nephropathy by suppressing connective tissue growth factor (CTGF) and receptor for advanced glycation endproducts (RAGE) gene expression in diabetic rats with streptozotocin.

Diabetic nephropathy is one of the major complications of diabetes, the use of medicinal plants is increasing due to fewer side effects. This study was designed to examine antidiabetic effects of Allium jesdianum (A.jesdianum) ethanolic extract and evaluate its effects on oxidative stress markers and the expression of connective tissue growth factor (CTGF) and receptor for advanced glycation endproducts (RAGE) genes in the kidney of type 1 diabetic rats. In this study, we randomly divided 24 rats into four groups with six rats in each group as follows: Cnt group: normal control receiving normal saline, Dibt group: diabetic control receiving normal saline daily, Dibt+A.jesdianum 250 group: diabetic rats receiving A.jesdianum at a dose of 250mg/kg bw daily, Dibt+A.jesdianum 500 group: diabetic rats receiving A.jesdianum at a dose of 500mg/kgbw daily. To induce diabetes, we used 55mg/kgbw dose of streptozotocin intraperitoneally. The concentration of fasting blood glucose (FBG) and serum urea, creatinine and albumin, SOD, MDA (using spectrophotometric methods) and gene expression of CTGF and RAGE in kidney tissue (using real-time PCR methods) were quantified in the diabetic rats that received A.jesdianum for 42days, and were compared to control rats. The results showed that in the diabetic group the FBG and serum urea, creatinine and expression of kidney CTGF and RAGE genes and the levels of SOD and MDA significantly increased and serum albumin significantly decreased compared to the Cnt group (p<0.001). Administration of A.jesdianum significantly improved the FBG and serum urea, creatinine and albumin compared to Dibt group (p<0.05). It was shown the A.jesdianum significantly decrease the kidney expression levels of CTGF and RAGE genes and improve oxidative stress (increased SOD and decreased MDA) in the kidney tissues when compared to Dibt group (p<0.001). Also, it was found that the beneficial effects of the A.jesdianum were dose-dependent. The results of this study showed that administration of A.jesdianum for 42days has beneficial anti-diabetic and anti-nephropathic effects in diabetic rats and can be used as an adjunct therapy in the treatment of diabetes.

CIHH protects the heart against left ventricular remodelling and myocardial fibrosis by balancing the renin-angiotensin system in SHR

AimThe aim of our study was to clarify the cardioprotection of chronic intermittent hypobaric hypoxia (CIHH) and the underlying mechanism in spontaneously hypertensive rats (SHR). Main methodsAdult male rats were divided into normal blood pressure Wistar-Kyoto rats (WKY) control (WKY-CON), WKY rats with CIHH treatment (WKY-CIHH), SHR control (SHR-CON) and SHR with CIHH treatment (SHR-CIHH) groups. SHR-CIHH and WKY-CIHH rats were subjected to hypobaric hypoxia simulating 4000-m altitude for 35 days, 5 h per day. Arterial blood pressure and cardiac function parameters, including ejection fraction, fractional shortening and left ventricular (LV) wall thickness, were evaluated. Cardiac pathomorphology and myocardial fibrosis were determined. The expression of angiotensin-converting enzyme (ACE), ACE2, Ang II, Ang1-7, AT1 receptor, Mas receptor, IL-6, TNF-α，IL-10, SOD and MDA were assayed in myocardium. Key findingsCIHH significantly decreased arterial blood pressure, alleviated LV hypertrophy, and improved cardiovascular function in SHR (P < 0.05–0.01). Also, CIHH protected SHR heart against morphological changes and fibrosis. In addition, CIHH significantly down-regulated the ACE/Ang II/AT1 receptor axis and up-regulated the ACE2/Ang1–7/Mas axis of renin-angiotensin system (RAS) in SHR (P < 0.05–0.01). CIHH significantly reduced IL-6, TNF-α, and MDA levels, but increased IL-10 and SOD in SHR myocardium (P < 0.05–0.01). SignificanceThe CIHH treatment protected the heart of SHR against LV remodelling and myocardial fibrosis, which might be carried out through a balance in the ACE/Ang II/AT1 axis and the ACE2/Ang1–7/Mas axis of the RAS to reduce inflammation, and inhibit oxidative stress.

Angiotensin Ⅱ inhibits AMPK/SIRT1 pathway by inducing oxidative stress in RAW264.7 macrophages

To investigate the mechanism by which angiotensin Ⅱ-induced oxidative stress response inhibits AMPK/ SIRT1 signaling in RAW264.7 macrophages. RAW264.7 cells were treated with 0.5, 1, 3, 10, or 20 μmol/L angiotensin Ⅱ for 24 h, and the changes in the expressions of AMPK, p-AMPK, and SIRT1 proteins were detected using Western blotting. The intracellular ROS release level was measured and the levels of SOD and MDA were detected. The effects of angiotensin Ⅱ type 1 receptor (AT1R) gene silencing on the cell response to angiotensin Ⅱ treatment were examined by detecting the changes in AMPK, p-AMPK and SIRT1 protein levels. The effects of a ROS inhibitor on cellular AMPK and SIRT1 were also examined. Angiotensin Ⅱ stimulation at 20 μmol/L significantly inhibited the phosphorylation of AMPK protein and increased cellular ROS release (P < 0.05). Treatment with 0.5-10 μmol/L angiotensin Ⅱ did not cause significant changes in SOD activity or MDA expression, but angiotensin Ⅱ at the dose of 20 μmol/L significantly inhibited SOD activity in the cells (P < 0.05). In the macrophages with AT1R gene silencing, treatment with angiotensin Ⅱ did not obviously inhibit AMPK phosphorylation or down- regulate SIRT1 expression. In cells treated with the ROS inhibitor, angiotensin Ⅱ failed to lower the level of AMPK phosphorylation or the expression of SIRT1. Angiotensin Ⅱ induces oxidative stress to cause disturbance of AMPK/ SIRT1 signaling pathway in macrophages.

Sığır Papillom ve Fibropapillomlarda Oksidatif Stres Kaynaklı DNA Hasarı ve Lipid Peroksidasyonun İmmunohistokimyasal Olarak Araştırılması

In this study, it was aimed to evaluate the expressions of 8-OHdG and MDA immunohistochemically in order to determine the oxidative stress-induced DNA damage and lipid peroxidation in 16 papillomas and 14 fibropapillomas from 30 cattle brought to our department between 2013-2020. Biopsy samples taken after surgery were fixed in buffered 10% formaldehyde solution. Sections of 5 μm thickness were taken from the paraffin blocks prepared after routine tissue follow-up procedures and Hematoxylin &amp;amp; Eosin staining was applied to the sections in order to detect histopathological changes. Avidin Biotin Peroxidase method was used for immunohistochemical staining. In papilloma cases, severe hyperkeratosis, retepects extending from the epidermis to the dermis, spongiosis and balloon-like degeneration in squamous epithelial cells, basophilic inclusion bodies in granular cells, ulcerations in the epidermis layer, hemorrhagic areas and an increase in kerato hyaline granules were observed. In addition to these findings, dense connective tissue increases were detected in fibropapilloma cases. In immunohistochemical evaluations, positive reactions for BPV were observed in the nuclei of cells in stratum granulosum. 8-OHdG positive reactions were detected in the cytoplasm and nucleus of the epidermal cells in papilloma cases, whereas in cases of fibropapilloma, reactions in these cells were observed in the cytoplasm of fibrocytes and fibroblasts in the dermis. Membranous MDA positive reactions were observed in the epidermal cells in papilloma cases, whereas MDA expressions were detected in the cytoplasm of fibrocytes and fibroblasts in the dermis of fibropapilloma cases. As a result of the literature reviews, no study data was found in which DNA damage due to oxidative stres and lipid peroxidation was detected by means of 8-OHdG and MDA expressions in papilloma and fibropapilloma cases of cattle, it is thought that the findings obtained in this study will contribute to the literature. In addition, it has been concluded that oxidative stress plays an important role in the pathogenesis of this tumor.

Immunohistochemical Investigation of Lipid Peroxidation in Renal Coccidiosis of Geese

Objective: In this study, we aimed to evaluate the oxidative damage caused by lipid peroxidation due to renal coccidiosis by histopathological and immunohistochemical methods.Material-Method: The material of this study was made up of tissue samples taken from 139 geese whose average age was 10 weeks, who were brought to our department dead between 2013-2020. Tissue samples taken were fixed in 10% buffered formaldehyde solution. 5 µm-thick sections were taken from the paraffin blocks prepared after routine tissue follow-up procedures. Hematoxylin &amp;amp; Eosin staining was applied to the sections in order to detect histopathological changes. Sections were examined and photographed under a light microscope. Results: Various clinical signs such as fever, respiratory distress, weakness, anorexia, tremors, inability to get up from the ground, balance disorders, rotational movement, diarrhea, wheezing were detected in geese. In systemic necropsies of geese, large and small white nodular structures were detected in the kidney. In histopathological examinations, coccidiosis agents (E. truncata) were found in the tubular epithelium of the kidney. Necrosis and mononuclear cell infiltration were observed in the tubules due to the presence of E. truncata. In addition, edema and hyperemia in the lungs, multifocal necrosis in the liver, cell infiltration in the portal spaces, and enteritis were other important histopathological findings. In one case, aspergillosis was detected together with renal coccidiosis. We observed that MDA expression was more severe in the oocyst stages, which is the mature form of the parasite, compared to other parasitic life stages. Conclusion: Based on the results obtained from this study, it was revealed that renal coccidiosis in geese caused lipid peroxidation / oxidative damage through the increase in MDA expression.

Azithromycin inhibited oxidative stress and apoptosis of high glucose‐induced podocytes by inhibiting STAT1 pathway

Azithromycin (AZM) has a therapeutic effect on diabetes, but there is no report on whether AZM has a therapeutic effect on diabetic nephropathy (DN) and its specific mechanism. Cell survival was detected by CCK-8. The expression of the inflammatory factors TNF-α, IL-1β, and IL-6 was determined by ELISA. The expression of inflammatory proteins MCP-1, NLPR3, and ASC was detected by western blot. The expression of MDA, LDH, and SOD was detected by the appropriate kit. Apoptosis was detected by flow cytometry and apoptosis-related proteins Bcl-2, Bax, Caspase-3, 6, 9, and Cleaved caspase-3, 6, 9 were detected by western blot. In addition, the expression of STAT1 was detected by western blot. AZM can increase the activity of high glucose-induced podocytes (p < .05). After high glucose induction, the expression of TNF-α, IL-1β, and IL-6 was increased and the expression of MCP-1, NLPR3, and ASC proteins was also increased (p < .001). When AZM was added, the expression of all the above-mentioned proteins was decreased (p < .001). In addition, MDA, LDH, and SOD were increased after high glucose induction, while decreased after AZM treatment (p < .001). AZM can inhibit apoptosis and the expression of Bax and Cleaved caspase-3, 6, 9, and promote the expression of Bcl-2 (p < .001). Furthermore, the expression of STAT1 was increased after high glucose induction, while the expression of STAT1 was decreased after AZM action (p < .01). By adding a STAT1 agonist IFN-γ, the effects of AZM on inflammation, oxidative stress, and apoptosis of high glucose-induced podocytes were inhibited (p < .05). AZM inhibited inflammation, oxidative stress, and apoptosis of high glucose-induced podocytes by inhibiting STAT1 pathway.

Hydrogen gas inhalation alleviates myocardial ischemia-reperfusion injury by the inhibition of oxidative stress and NLRP3-mediated pyroptosis in rats

AimsReperfusion therapy is the most common and effective treatment against ischemic heart disease (IHD), but the process inflicts massive ischemia/reperfusion (I/R) injury for which no treatment exists. Notably, reperfusion after ischemia causes ischemia/reperfusion injury (IR injury) and the “no-reflow” phenomenon seriously affecting the therapeutic effects in clinical practice. The principle purpose of this study is to validate the effect of hydrogen gas on IHD and further explore the mechanism of hydrogen gas in alleviating myocardial I/R injury and no-reflow phenomenon. Materials and methodsThe rat model of myocardial ischemia-reperfusion was well established. Myocardial infarct size was evaluated by TTC & Evans blue staining. The no-reflow area and the cardiac function were assessed by thioflavin-S staining and echocardiography respectively. Microstructure and mitochondria of myocardial tissue were assessed by transmission electron microscope. Western blot and immunohistochemistry were used to evaluate the expression of NLRP3 mediated pyroptosis related proteins. The 8-OHdG, MDA and serum total ROS were used to evaluate the degree of oxidative stress. Key findingsThe myocardial infarct size, no-reflow area, cardiac function, microstructure and mitochondrial morphology of I/R model rats were significantly improved after hydrogen inhalation. In addition, the expression of 8-OHdG, MDA, ROS and NLRP3 mediated pyroptosis related proteins were significantly decreased. SignificanceWe found that oxidative stress and NLRP3 mediated pyroptosis are the important mechanisms for hydrogen to alleviate myocardial I/R injury, and we also confirmed that hydrogen can significantly improve no reflow phenomenon caused by ischemia-reperfusion.