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ObjectiveBased on the nuclear factor erythroid 2 related factor 2 （Nrf2）/heme oxygenase-1 （HO-1） signaling pathway， this paper explores the effect of Sinisan （SNS） on liver oxidative stress injury in cholestatic hepatitis rats and its mechanism. MethodThirty 6-week-old male SD rats were randomly divided into a control group， model group， low and high dose groups of SNS （2.5 and 5 g·kg-1） and ursodeoxycholic acid group （UDCA， 63 mg·kg-1）， with six rats in each group. Rats were administrated for seven consecutive days. On the 5th day， the control group was given olive oil of 10 mL·kg-1， and the other groups were given alpha-naphthalene isothiocyanate （ANIT） of 80 mg·kg-1. The serum biochemical indicator levels of cholestasis and the content of antioxidant factors in rat liver were detected by enzyme-linked immunosorbent assay （ELISA）. Hematoxylin-eosin （HE） staining was used to observe the pathological changes in liver tissue. The relative mRNA and protein expressions of Nrf2， HO-1， and quinone oxidoreductase 1 （NQO1） in liver tissue were detected by real-time fluorescence quantitative polymerase chain reaction （Real-time PCR） and Western blot. ResultCompared with the control group， the model group showed a significant increase in the serum biochemical indicator levels of cholestasis and the content of antioxidant factors in liver tissue （P<0.01）. There were obvious pathological changes in the model group such as the disordered arrangement of hepatocytes， obvious congestion and necrosis in the portal area， infiltration of inflammatory cells， and destruction of the interlobular bile duct. The relative mRNA and protein expressions of Nrf2， HO-1， and NQO1 in liver tissue were significantly down-regulated in the model group （P<0.05， P<0.01）. Compared with the model group， the groups of SNS showed a significant decrease in the serum biochemical indicator levels of cholestasis and the content of antioxidant factors in liver tissue （P<0.01）， and the pathological liver injury was obviously improved. The necrotic area was reduced， and the infiltration of inflammatory cells was decreased. In addition， there was a small amount of extravasated blood in the interlobular vein. The relative mRNA and protein expressions of Nrf2， HO-1， and NQO1 in liver tissue were significantly up-regulated （P<0.05， P<0.01）. ConclusionSNS can significantly improve liver injury in cholestatic hepatitis rats， and its mechanism may be related to the inhibition of oxidative stress response mediated by the Nrf2/HO-1 signaling pathway.
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ObjectiveTo investigate the effect of linalool against acute liver injury induced by aflatoxin B1（AFB1） in rats and explore its protective mechanism. MethodTwenty male SPF SD rats were randomly divided into three groups： Control （n=6）， AFB1 （n=7）， and linalool （n=7） groups. Linalool solution （200 mg·kg-1） was administered preventatively for 14 days， while the control and AFB1 groups intragastrically received an equivalent volume of double distilled water. After preventative administration of linalool， AFB1 solution （1 mg·kg-1， dissolved in saline） was intraperitoneally injected for two consecutive days to induce acute liver injury in rats. Samples were collected and processed 14 days after model establishment. Pathological changes in liver tissue of rats were observed using Hematoxylin-eosin（HE） staining and Masson staining. Biochemical detection was performed to measure the levels of alanine transaminase（ALT）， aspartate transaminase（AST）， γ-glutamyl transferase（GGT）， lactate dehydrogenase（LDH）， alkaline phosphatase（ALP）， total bilirubin（TBil）， direct bilirubin（DBil）， indirect bilirubin（IBil）， malondialdehyde（MDA）， superoxidedismutase（SOD）， catalase（CAT） ， glutathione（GSH）， Fe3+， and Fe2+ in the liver tissue. Western blot was adopted to assess protein expression levels of nuclear factor-erythroid 2-related factor 2（Nrf2） and heme oxygenase-1（HO-1）. Molecular docking was performed to verify the binding between linalool and key proteins of the Nrf2/HO-1 signaling pathway. Molecular dynamics techniques were used to confirm the stability and affinity of linalool binding with key proteins of the Nrf2/HO-1 signaling pathway. ResultPathological results showed that compared to that in the AFB1 group， the liver structure in the linalool group tended to be normal， with a significant decrease in blue collagen fibers. The linalool group exhibited significantly reduced levels of ALT， AST， GGT， LDH， ALP， TBil， DBil， and IBil （P<0.01）， Fe3+ and Fe2+ content， and oxidative stress marker MDA （P<0.01）. The levels of antioxidants SOD， CAT， and GSH significantly increased （P<0.01）. Molecular docking showed a molecular docking energy between linalool and Nrf2 and HO-1 targets of -5.495 6 and -5.199 4 kcal·mol-1（1 cal≈4.186 J）， respectively. Molecular dynamics results indicated strong affinity in the binding of linalool with Nrf2 and HO-1. Western blot revealed a significant increase in Nrf2 protein expression （P<0.05） and a decrease in HO-1 protein expression （P<0.01） in the linalool group. ConclusionLinalool may protect against AFB1-induced acute liver injury by modulating the Nrf2/HO-1 ferroptosis signaling pathway to inhibit liver cell ferroptosis and regulate hepatic oxidative stress levels.
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ObjectiveTo investigate the effect and mechanism of Zhenwutang on renal oxidative damage in the mouse model of diabetic kidney disease with the syndrome of spleen-kidney Yang deficiency via the nuclear factor erythroid 2-related factor-2 （Nrf2）/heme oxygenase-1 （HO-1）/glutathione peroxidase 4 （GPX4） signaling pathway. MethodTwenty-five 7-week-old SPF-grade male db/m mice and 95 7-week-old SPF-grade male db/db mice were adaptively fed for a week. A blank group was set with the db/m mice without treatment， and the other mice were administrated with Rhei Radix et Rhizoma decoction and hydrocortisone for the modeling of diabetic kidney disease with the syndrome of spleen-kidney Yang deficiency. The modeled mice were randomized into the model， irbesartan （25 mg·kg-1）， and high-， medium-， low-dose （33.8， 16.9， 8.45 g·kg-1） Zhenwutang groups （n=15） and administrated with corresponding drugs for 8 weeks. The survival status of mice was observed， and the traditional Chinese medicine （TCM） syndrome score was recorded. The indicators related to spleen-kidney Yang deficiency， fasting blood glucose （FBG）， and renal function indicators were determined. Hematoxylin-eosin staining was employed to observe the histopathological changes of the renal tissue in each group. Biochemical kits were used to determine the oxidative stress-related indicators in the renal tissue. Real-time polymerase chain reaction and Western blotting were employed to determine the mRNA and protein levels， respectively， of Nrf2， HO-1， glutamate-cysteine ligase catalytic subunit （GCLC）， and GPX4 in the renal tissue of mice in each group. ResultCompared with the blank group， the modeling increased the TCM syndrome score （P<0.05）， elevated the estradiol （E2） and FBG levels （P<0.05）， lowered the testosterone （T）， triiodothyronine （T3）， and tetraiodothyronine （T4） levels （P<0.05）， and weakened the renal function （P<0.05）. In addition， the modeling led to glomerular hypertrophy and glomerular mesangial and basal thickening， decreased the catalase （CAT） activity， total antioxidant capacity （T-AOC）， and glutathione （GSH） content （P<0.05）， increased the malondialdehyde （MDA） content （P<0.05）， and down-regulated the mRNA and protein levels of Nrf2， HO-1， GCLC， and GPX4 in the renal tissue （P<0.05）. Compared with the model group， high and medium doses of Zhenwutang decreased the TCM syndrome score and E2 content （P<0.05）， increased the T， T3， and T4 content （P<0.05）， improved the renal function （P<0.05）， alleviated the pathological changes in the renal tissue， increased CAT， T-AOC， and GSH （P<0.05）， reduced MDA （P<0.05）， and up-regulated the mRNA and protein levels of Nrf2， HO-1， GCLC， and GPX4 in the renal tissue （P<0.05）. ConclusionZhenwutang can improve the general state and renal function and reduce the oxidative damage and pathological changes in the renal tissue of db/db mice with spleen-kidney Yang deficiency by regulating the Nrf2/HO-1/GPX4 signaling pathway.
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ObjectiveTo investigate the effect and mechanism of Shenqi Tangluo pill （SQTLP） on oxidative stress injury of skeletal muscle of type 2 diabetes mellitus （T2DM） mice based on nuclear factor erythroid 2-related factor 2 （Nrf2）/heme oxygenase-1 （HO-1）/NAD（P）H quinone oxidoreductase 1 （NQO1） pathway. MethodA total of 60 7-week-old male db/db mice ［specific pathogen-free （SPF） grade］ were selected and fed for one week for adaption. They were divided into the model control group， SQTLP low-， medium- and high-dose （19， 38， and 76 g·kg-1） groups and metformin group （0.26 g·kg-1） by gavage. Each group consisted of 12 mice. Twelve male db/m mice of the same age were selected as the blank group. The intervention was implemented continuously for 8 weeks. Fasting blood glucose （FBG） was detected. Fasting serum insulin （FINS） levels were detected by enzyme-linked immunosorbent assay （ELISA）， and the homeostasis model assessment-insulin resistance （HOMA-IR） index and the homeostasis model assessment-insulin sensitivity index （HOMA-ISI） were calculated. Oral glucose tolerance test （OGTT） and insulin tolerance test （ITT） were conducted. The activities of superoxide dismutase （SOD） and glutathione peroxidase （GSH-Px） and the contents of malondialdehyde （MDA） and reduced nicotinamide adenine dinucleotide phosphate （NADPH） in skeletal muscle tissues were detected by biochemical kits. Hematoxylin-eosin （HE） staining was used to observe the pathological changes in skeletal muscle tissues. The levels of reactive oxygen species （ROS） and 4-hydroxynonenal （4-HNE） in skeletal muscle tissue were detected by immunofluorescence （IF）. The expression levels of Nrf2， HO-1， NQO1 and glutamate-cysteine ligase catalytic subunit （GCLC） proteins in skeletal muscle tissues were detected by Western blot. ResultCompared with those in the blank group， FBG， FINS and HOMA-IR in the model group were significantly increased （P<0.05）， while HOMA-ISI was decreased （P<0.05）. The results of OGTT and ITT showed that blood glucose was significantly increased at all time points （P<0.05）， and glucose tolerance and insulin tolerance were significantly impaired. SOD and GSH-Px activities in skeletal muscle tissues were significantly decreased （P<0.05）， and MDA and NADPH contents were significantly increased （P<0.05）. In skeletal muscle tissues， the arrangement of muscle fibers was loose， the nucleus was disordered， and inflammatory cells were infiltrated. The expression levels of ROS and 4-HNE in skeletal muscle tissues were significantly increased （P<0.05）. The protein expression levels of Nrf2， HO-1， NQO1 and GCLC in skeletal muscle tissues were significantly decreased （P<0.05）. Compared with those in the model group， FBG， FINS and HOMA-IR in the metformin group were significantly decreased （P<0.05）， while HOMA-ISI was increased （P<0.05）. The results of OGTT and ITT showed that blood glucose in the metformin group was significantly decreased at all time points （P<0.05）. The activities of SOD and GSH-Px in skeletal muscle tissues were significantly increased （P<0.05）， while the contents of MDA and NADPH were significantly decreased （P<0.05）. No obvious abnormality was found in the skeletal muscle tissue of the metformin group. The expressions of ROS and 4-HNE in skeletal muscle tissues were decreased （P<0.05）. The protein expression levels of Nrf2， HO-1， NQO1 and GCLC in skeletal muscle tissues were significantly increased （P<0.05）. Compared with those in the model group， FBG， FINS and HOMA-IR in the SQTLP medium- and high-dose groups were significantly decreased （P<0.05）， while HOMA-ISI was increased （P<0.05）. The results of OGTT and ITT showed that the glucose tolerance and insulin tolerance of mice were improved in each dose group of SQTLP. The GSH-Px activity in the SQTLP low-dose group was significantly increased （P<0.05）， and the NADPH content was decreased （P<0.05）. The activities of SOD and GSH-Px in the SQTLP medium- and high-dose groups were significantly increased （P<0.05）， while the contents of MDA and NADPH were significantly decreased （P<0.05）. The skeletal muscle tissue injury of mice in each dose group of SQTLP was ameliorated to different degrees. In the SQTLP medium- and high-dose groups， the expressions of ROS and 4-HNE were decreased （P<0.05）， and the protein expression levels of Nrf2， HO-1， NQO1 and GCLC were significantly increased （P<0.05）. Compared with those in the SQTLP low-dose group， FBG and HOMA-IR in the SQTLP high-dose group were significantly decreased （P<0.05）， while HOMA-ISI was increased （P<0.05）. The results of OGTT and ITT showed that the SQTLP high-dose group significantly improved the glucose tolerance and insulin tolerance of mice. The activities of SOD and GSH-Px in skeletal muscle tissues were significantly increased （P<0.05）， while the contents of MDA and NADPH were significantly decreased （P<0.05）. No obvious abnormality was found in the skeletal muscle tissue， the expressions of ROS and 4-HNE were decreased （P<0.05）， and the protein expression levels of Nrf2， HO-1， NQO1 and GCLC were significantly increased （P<0.05） in the skeletal muscle tissue of the SQTLP high-dose group. ConclusionSQTLP can significantly improve IR in T2DM mice， and the mechanism is related to SQTLP activating the Nrf2/HO-1/NQO1 signaling pathway， promoting the expression of antioxidant enzymes， and thus improving the oxidative stress injury in the skeletal muscle.
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Myocardial infarction （MI） is a common cardiovascular disease in clinical practice and one of the main causes of cardiovascular mortality. Its pathogenesis is complex and associated with oxidative stress reactions. Nuclear factor E2-related factor 2 （Nrf2） is a key factor in regulating oxidative stress reactions. It can regulate the expression of heme oxygenase-1 （HO-1）， playing a role in maintaining the oxidative-reductive homeostasis in the body. During the course of MI， the biological activity and levels of Nrf2 and HO-1 decrease， leading to weakened tissue antioxidant and anti-inflammatory capabilities， endothelial damage in myocardial blood vessels， release of vascular cell adhesion factors， and impaired endothelial function. In recent years， many basic research studies have explored the role and mechanisms of traditional Chinese medicine （TCM） in treating MI by modulating the Nrf2/HO-1 signaling pathway. The results have indicated that the Nrf2/HO-1 signaling pathway is an important potential target for TCM in the treatment of MI. This article reviewed the mechanism of the Nrf2/HO-1 signaling pathway in MI and the research progress of TCM in targeting and regulating this pathway， aiming to provide a theoretical basis for the prevention and treatment of MI and further drug development.
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ObjectiveTo investigate the effect of Glycyrrhizae Radix et Rhizoma （GR）-containing serum on lipopolysaccharide （LPS）-induced inflammation in human colon epithelial adenocarcinoma cells （Caco2） based on inhibition of ferroptosis by the nuclear factor erythroid 2-related factor 2 （Nrf2）/heme oxygenase-1 （HO-1） pathway. MethodCaco2 cells were divided into a normal group， a model group （LPS， 200 μg·L-1）， low-， medium-， and high-dose GR-containing serum groups （5%， 10%， 20%）， and a ferroptosis inhibitor group （3-amino-4-cyclohexylamino-benzoic acid ethyl ester， Fer-1， 10 μmol·L-1）. The cells in the normal group were cultured normally， while those in other groups underwent the induction of an inflammation model. The cells in the low-， medium-， and high-dose GR-containing serum groups were treated with 5%， 10%， and 20% GR-containing serum for 24 hours， respectively， and the cells in the ferroptosis inhibitor group were treated with Fer-1 for 24 hours. Transmission electron microscopy was used to observe mitochondrial morphology in each group. Flow cytometry was used to detect intracellular Fe2+ levels. Microplate assays were performed to measure superoxide dismutase （SOD） activity， malondialdehyde （MDA） and glutathione peroxidase （GSH-Px） levels. Enzyme-linked immunosorbent assay （ELISA） was used to measure interleukin-1β （IL-1β）， IL-6， IL-10， and tumor necrosis factor-α （TNF-α） levels. Western blot was used to measure the expression levels of Nrf2， HO-1， ferritin heavy chain 1 （FTH1）， and glutathione peroxidase 4 （GSH-Px4） proteins. Small interfering RNA （siRNA） was used to investigate the role of Nrf2 in ferroptosis regulation. The cells after interference were divided into a negative control （NC） group， a Si-Nrf2 group， a GR-containing serum （20%） + Si-Nrf2 group， and a GR-containing serum （20%） + NC group. Microplate assays were performed to measure MDA， SOD， and GSH-Px levels， and Western blot was used to measure the expression levels of Nrf2， HO-1， FTH1， and GSH-Px4 proteins. ResultCompared with the normal group， the model group showed mitochondrial contraction， increased mitochondrial membrane thickness， and smaller mitochondrial morphology， increased Fe2+ content （P<0.01）， blunted SOD activity （P<0.01）， decreased GSH-Px expression （P<0.01）， increased MDA content （P<0.01）， reduced expression levels of Nrf2 and HO-1 （P<0.05）， reduced FTH1 expression （P<0.01）， and down-regulated GSH-Px4 expression （P<0.01）. In the GR-containing serum groups， the medium- and high-dose groups showed a significant decrease in Fe2+ content （P<0.01）， potentiated SOD and GSH-Px activities （P<0.01）， and decreased MDA levels （P<0.01）. The high-dose group showed a significant increase in Nrf2 expression （P<0.05）， and the medium-dose group showed increased expression of HO-1 and GSH-Px4 proteins （P<0.05）. The expression levels of FTH1 significantly increased in the low-， medium-， and high-dose groups （P<0.01）. The study on mechanism revealed that compared with the NC group， the cells transfected with Nrf2 siRNA showed increased MDA content （P<0.01）， blunted SOD activity （P<0.01）， decreased GSH-Px activity （P<0.01）， decreased expression of Nrf2 and HO-1 （P<0.01）， and reduced levels of FTH1 and GSH-Px4 proteins （P<0.01）. Compared with the Si-Nrf2 group， the cells treated with GR-containing serum showed a decrease in MDA content （P<0.01）， an increase in SOD activity （P<0.01）， an increase in GSH-Px activity （P<0.01）， increased expression of Nrf2 and FTH1 proteins （P<0.05）， and higher expression levels of HO-1 and GSH-Px4 proteins （P<0.01）. ConclusionGR-containing serum can reduce the inflammatory cytokines and oxidative stress levels in LPS-induced Caco2 cells. Its mechanism is related to the promotion of Nrf2/HO-1 signaling pathway expression， alleviating intracellular lipid peroxidation and inhibiting ferroptosis.
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AIM:To analyze the correlation between serum nesfatin-1, apelin and heme oxygenase-1(HO-1)levels and the severity of diabetic retinopathy(DR).METHODS:Totally 100 patients with type 2 diabetes mellitus(T2DM)who were admitted to the hospital from September 2020 to September 2022 were selected. They were divided into non-DR(NDR)group(35 cases), nonproliferative DR(NPDR)group(33 cases)and proliferative DR(PDR)group(32 cases)according to the condition of fundus lesions. Another 30 healthy individuals who received health check-ups in the hospital during the same period were selected as the control group. Serum nesfatin-1, apelin and HO-1 levels in each group were detected, and panretinal ischemia index(ISI)was evaluated.RESULTS:Serum nesfatin-1 and HO-1 levels in the T2DM patients were lower, and apelin level was higher as compared with the control group. The levels of nesfatin-1 and HO-1 in the PDR group were the lowest, while the apelin level was the highest. Panretinal ISI in the PDR group was higher than that in the NPDR group(4.56±0.57 vs. 2.05±0.29, P&#x003C;0.05). Correlation analysis found that serum nesfatin-1 and HO-1 levels were negatively correlated with panretinal ISI in patients with DR, while apelin level was positively correlated with panretinal ISI. The receiver operator characteristic(ROC)curve analysis found that the areas under the curves of serum nesfatin-1, apelin and HO-1 for predicting PDR were 0.842, 0.833 and 0.807 respectively.CONCLUSION:Serum nesfatin-1, apelin and HO-1 levels are closely related to the severity of DR. Dynamic monitoring of serum nesfatin-1, apelin and HO-1 levels is important for the early detection of PDR.
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ObjectiveTo study the effect of Buyang Huanwutang on Kelch-like Ech-related protein 1 （Keap1）/nuclear factor E2-related factor 2 （Nrf2）/heme oxygenase-1 （HO-1） antioxidant signaling pathway in rats with idiopathic pulmonary fibrosis （IPF） and explore the mechanism of this prescription in the treatment of IPF. MethodForty SPF-grade male SD rats were assigned into a sham operation group， a model group， a Buyang Huanwutang group， and a nintedanib group according to random number table method， with 10 rats in each group. IPF rat model was established by intratracheal infusion of bleomycin （0.005 g·kg-1） in other groups except the sham operation group. Buyang Huanwutang group was administrated with Buyang Huanwutang (14.84 g·kg-1)，intragastric administration of nitedanib suspension (0.1 g·kg-1)，sham operation group and model group were given equal volume of normal saline， for 28 days. After lung function test， serum and lung tissue samples were collected. Hematoxylin-eosin （HE） staining and Masson trichrome staining were employed to observe the pathological changes of the lung tissue. The content of hydroxyproline （HYP） in lung tissue was detected. The levels of malondialdehyde （MDA） in serum and lung tissue， and the activities of superoxide dismutase （SOD）， glutathione peroxidase （GSH-Px） and catalase （CAT） were determined. The mRNA and protein levels of Keap1， Nrf2， and HO-1 was determined by Real-time fluorescent quantitative polymerase chain reaction， immunohistochemical staining， and Western blot. ResultCompared with the sham operation group， the modeling increased the resistance and elasticity and decreased the compliance of respiratory system （P<0.01）， elevated the lung index， pathological score， and HYP content in lung tissue （P<0.01）， and enriched MDA in serum and lung tissue， while it decreased the activities of SOD， GSH-Px， and CAT （P<0.01）. Furthermore， the modeling down-regulated the mRNA and protein levels of Keap1 and up-regulated those of Nrf2 and HO-1 in lung tissue （P<0.01）. Compared with the model group， Buyang Huanwutang decreased the resistance and elasticity and increased the compliance of respiratory system （P<0.01）， lowered the lung index， pathological score， and HYP content in lung tissue （P<0.01）， and reduced MDA in serum and lung tissue， while it increased the activities of SOD， GSH-Px， and CAT （P<0.01）. Additionally， Buyang Huanwutang down-regulated the expression of Keap1 and up-regulated that of Nrf2 and HO-1 in lung tissue （P<0.05， P<0.01）. ConclusionBuyang Huanwutang can activate Keap1/Nrf2/HO-1 signaling pathway to enhance the antioxidant capacity and slow down the pathological process of IPF in rats.
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Osteoporosis is a chronic skeletal disease characterized by low bone mass， destruction of bone tissue microarchitecture， and imbalance of bone homeostasis， leading to increased bone fragility and increased risk of fractures. Oxidative stress caused by the disruption of the balance between excess reactive oxygen species （ROS） and the anti-oxidative system is an important factor in the occurrence and progression of osteoporosis. Nuclear factor E2-related factor 2 （Nrf2）/heme oxygenase-1 （HO-1） is an important anti-oxidative stress pathway. Nrf2 is a primary factor in regulating cellular oxidative stress. Activating Nrf2 can stimulate the expression of HO-1. HO-1 is a key enzyme whose metabolites are bile green Oxygen， carbon monoxide， and free iron. The metabolites can scavenge ROS， thereby exerting an antioxidant effect in cells. At present， domestic and foreign scholars have reported that the Nrf2/HO-1 signaling pathway is closely related to the occurrence and development of osteoporosis and the mechanism of drugs. Chinese medicine can effectively solve the insufficiency of western medicine with multi-target， multi-channel， and multi-level advantages. Chinese medicine can resist oxidative stress， inflammatory response， and apoptosis by regulating the Nrf2/HO-1 signaling pathway， thus treating osteoporosis. This article reviewed the relationship between Nrf2/HO-1 signaling pathway and its key target protein factors and osteoporosis， to clarify the important role of the Nrf2/HO-1 signaling pathway in osteoporosis. At the same time， a systematic summary of Chinese medicines targeting and regulating the Nrf2/HO-1 signaling pathway for the treatment of osteoporosis was conducted， to provide a theoretical basis for further precise treatment of osteoporosis.
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This study explored whether Sagittaria sagittifolia polysaccharides(SSP) activates the nuclear factor erythroid-2-related factor2(Nrf2)/heme oxygenase-1(HO-1) signaling pathway to protect against liver damage jointly induced by multiple heavy metals. First, based on the proportion of dietary intake of six heavy metals in rice available in Beijing market, a heavy metal mixture was prepared for inducing mouse liver injury and HepG2 cell injury. Forty male Kunming mice were divided into five groups: control group, model group, glutathione positive control group, and low-and high-dose SSP groups, with eight mice in each group. After 30 days of intragastric administration, the liver injury in mice was observed by HE staining. In the in vitro experiment, MTT assay was conducted to detect the effects of SSP at 0.25, 0.5, 1, and 2 mg·mL~(-1) on HepG2 cell survival at different time points. The content of alanine transaminase(ALT) and aspartate aminotransferase(AST) in the 48-h cell culture fluid was measured using micro-plate cultivation method, followed by the detection of the change in reactive oxygen species(ROS) content by flow cytometry. The mRNA expression levels of Nrf2 and HO-1 in cells were determined by RT-PCR, and their protein expression by Western blot. HE staining results showed that compared with the model group, the SSP administration groups exhibited significantly alleviated inflammatory cell infiltration and fatty infiltration in the liver, with better outcomes observed in the high-dose SSP group. In the in vitro MTT assay, compared with the model group, SSP at four concentrations all significantly increased the cell survival rate, decreased the ALT, AST, and ROS content(P<0.05), and down-regulated Nrf2 and HO-1 mRNA and protein expression(P<0.05). SSP significantly improves inflammatory infiltration in the liver tissue of mice exposed to a variety of heavy metals and corrects the liver fat degeneration, which may be related to its regulation of the Nrf2/HO-1 signaling pathway, reduction of ROS, and alleviation of oxidative damage.
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Animais , Masculino , Camundongos , Heme Oxigenase-1/metabolismo , Fígado , Metais Pesados/metabolismo , Fator 2 Relacionado a NF-E2/metabolismo , Estresse Oxidativo , Polissacarídeos/farmacologia , RNA Mensageiro/metabolismo , Espécies Reativas de Oxigênio/metabolismo , Sagittaria/metabolismoRESUMO
Objective:To investigate the renoprotective effects that Sanjiao Qushi prescription ameliorates cationic bovine serum albumin （C-BSA） induced membranous nephropathy（MN） in mice model and its influence on nuclear factor erythroid-2-related factor-2（Nrf2）/heme oxygenase-1（HO-1） signaling pathway. Method:Sixty female BALB/c mice were randomly divided into the normal group（<italic>n</italic>=10） and the model group（<italic>n</italic>=50）. The mice in the model group received C-BSA injection via tail vein （6.5 mg·kg<sup>-1</sup>）. The mice that were successfully modeled were randomized into the model group， the low dose Sanjiao Qushi prescription group（3.71 g·kg<sup>-1</sup>）， the high dose Sanjiao Qushi prescription group（7.42 g·kg<sup>-1</sup>） and benazepril hydrochloride group（1.3 mg·kg<sup>-1</sup>）. And they were administered with the corresponding medicine by gavage once a day for four consecutive weeks. 24 hour-urine protein quantitation were performed before C-BSA injection and after C-BSA injection as well as the medicine gavage. When the treatment was finished， all of the mice were sacrificed and the biochemical indicators such as serum creatinine（SCr）， blood urea nitrogen（BUN）， triglyceride（TG）， total cholesterol（TC）， total protein（TP） and albumin（Alb） were measured. And the renal pathological morphology changes were observed by light microscope with hematoxylineosin（HE）， Masson and periodic acid-silver metheramine（PASM） staining. The deposition of immunoglobulin G（IgG） in the glomerulus was detected by fluorescence microscope. The expression of reactive oxygen species （ROS） of kidney was detected by fluorescence immunoassay. The protein expression levels of Nrf2 in cell nucleus and cytoplasm and the downstream protein factors HO-1 and NADH quinone acceptor oxidoreductase 1（NQO1） were detected by Western blot. Result:Compare to normal group， the levels of 24 hour-urine protein quantitation， TG and TC significantly increased in model group（<italic>P</italic><0.01）， while TP and Alb levels significantly decreased（<italic>P</italic><0.01）. The model group exhibited enlarged volume of glomerular， significantly thickened glomerular basement membrane（GBM）， fuchsinophilic protein deposition and spike formation through light microscope. Immunofluorescence staining for the model group exhibited granular deposition of IgG along the capillary wall. The expression of ROS in kidney significantly increased（<italic>P</italic><0.01）. The protein expression levels of Nrf2 in cell nucleus significantly increased（<italic>P</italic><0.01）， while Nrf2 in cytoplasm significantly decreased（<italic>P</italic><0.01）.The protein expression levels of HO-1 and NQO1 significantly increased（<italic>P</italic><0.01）. Compared to model group， the levels of 24 hour-urine protein quantitation， TG and TC significantly decreased in each treated group（<italic>P</italic><0.01）， TP and Alb levels significantly increased（<italic>P</italic><0.05，<italic>P</italic><0.01）. The pathological damages alleviated obviously. The expression of ROS in kidney significantly decreased（<italic>P</italic><0.01）. The protein expression levels of Nrf2 in cell nucleus significantly decreased（<italic>P</italic><0.01）， while Nrf2 in cytoplasm significantly increased（<italic>P</italic><0.05，<italic>P</italic><0.01）. The protein expression levels of HO-1 and NQO1 significantly increased（<italic>P</italic><0.01）. Conclusion:Sanjiao Qushi prescription worked on MN mice possibly by regulating related proteins in the Nrf2/HO-1 signaling pathway and relieving oxidative stress， thus decreasing 24 hour-urine protein and blood lipid， increasing serum protein， and alleviating the pathological damages to protect renal function and delay progress of the disease.
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Objective:To explore the inhibitory effect of dihydroartemisinin （DHA） on the proliferation of HepG2 cells， elucidate the mechanism from the perspectives of oxidative damage and energy metabolism， and discuss the possibility of combined use of DHA with sorafenib （Sora）. Method:Cell counting kit-8 （CCK-8） assay was used to obtain the 50% inhibitory concentration （IC<sub>50</sub>） of DHA and Sora on HepG2 and SW480 cells and Chou-Talalay method was used to obtain the combination index （CI） of DHA and Sora. HepG2 cells were classified into the control group， DHA group （10 µmol·L<sup>-1</sup>）， Sora group （5 µmol·L<sup>-1</sup>）， and DHA + Sora group （DHA 10 µmol·L<sup>-1</sup>， Sora 5 µmol·L<sup>-1</sup>） and then incubated with corresponding drugs for 8-12 h. Seahorse XF glycolytic rate assay kit and cell mito stress test kit were employed to respectively detect the glycolysis function of cells and oxidative phosphorylation function of mitochondria. DCFH-DA and lipid peroxidation MDA assay kit were separately used to analyze the intracellular levels of reactive oxygen species （ROS） and malondialdehyde （MDA）. Western blot was applied to determine the intracellular levels of heme oxygenase-1 （HO-1） and glutamate-cysteine ligase catalytic subunit （GCLC）. Result:Compared with the control group， DHA alone inhibited the ATP synthesis in mitochondrial oxidative phosphorylation and glycolysis （<italic>P</italic><0.01）， increased the levels of intracellular ROS and MDA （<italic>P<</italic>0.05）， and decreased the levels of HO-1 and GCLC （<italic>P<</italic>0.05） in HepG2 cells. DHA and Sora had synergistic inhibitory effect on proliferation of HepG2 and SW480 cells， with CI < 0.90. The DHA + Sora group showed stronger suppression of ATP synthesis in mitochondrial oxidative phosphorylation and glycolysis （<italic>P</italic><0.01）， higher levels of intracellular ROS and MDA （<italic>P<</italic>0.01）， and lower levels of intracellular antioxidation-related proteins HO-1 and GCLC in HepG2 cells （<italic>P<</italic>0.01） than the DHA group. Conclusion:DHA may increase the level of MDA by reducing HO-1 and GCLC and increasing ROS in HepG2 cells， which results in mitochondria oxidative damage， restricts cell glycolysis and mitochondrial oxidative phosphorylation， and thus finally inhibits the proliferation of HepG2 cells. DHA and Sora have synergistic inhibitory effect on the proliferation of HepG2 and SW480 cells， and the mechanism may be related to the synergistic oxidative damage that affects the mitochondrial electron transport chain and suppresses cell energy metabolism.
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Ganoderic triterpenoids (GTs) are the primary bioactive constituents of the Basidiomycotina fungus,
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Objective: To investigate the expression of NAD (P) H: quinone oxidoreductase 1 (NQO1) and heme oxygenase-1 (HO-1) in T-cell lymphoma (TCL), and investigate the correlation between these two indicators and other clinicopathological parameters in TCL. Methods: Clinical data of 61 patients with TCL who were initially diagnosed in Gansu Provincial Hospital were analyzed retrospectively. Immunohistochemical examination was performed to detect NQO1 and HO-1 expression levels in 61 TCL tissues (TCL group) and 20 lymph node reactive hyperplasia tissues (control group). Results: Positive expression rates of NQO1 and HO-1 were significantly higher in TCL tissues than in lymph node reactive hyperplasia tissues (P<0.05). NQO1 expression was closely related with Ann-Arbor clinical stage and B symptoms (P<0.05); HO-1 expression was correlated with clinical stage, bone marrow invasion, and B symptoms (P<0.05). NQO1 and HO-1 expression levels were not related to age, sex, lactate dehydrogenase level, and pathological type (P>0.05); there was a correlation between NQO1 and HO-1 expression (r=0.264; P=0.040). Conclusions: NQO1 and HO-1 are highly expressed in TCL and may interact and contribute to the occurrence and development of TCL.
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Objective To explore the roles of heme oxygenase-1 (HO-1 ) and protoporphyrin zinc IX (ZnPPIX ) , its inhibitor , in cisplatin chemotherapy for gastric cancer so as to provide potential targets for chemosensitivity in gastric cancer .Methods Gastric cancer cell line SGC7901 was used in vitro .MTT assay was carried out to determine the effects of ZnPPIX and CDDP on the proliferation in gastric cancer cells .The expression of HO-1 in gastric cancer cells was measured by Real-time PCR and Western blot ,respectively .The gastric cancer xenografts in nude mice were used to study the effects of ZnPPIX and CDDP in gastric cancer on tumor formation in vivo .Results The proliferation of cancer cells ,interfered by CDDP in combination with ZnPPIX ,could be significantly inhibited (P<0 .05) .Moreover ,CDDP could increase the expression of HO-1 in gastric cancer cells , which was reversed by ZnPPIX (P<0 .05) .The animal experiment showed that CDDP could inhibit gastric cancer growth in nude mice and reduce tumor volume and weight . Conclusion ZnPPIX could enhance the chemosensitivity of CDDP in gastric cancer ,which may be a potential sensitizer of cisplatin-based chemotherapy in gastric cancer .
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Nrf2/ARE pathway has become an attractive target using phytochemicals for the prevention and treatment of oxidative stress-related diseases including neurodegenerative diseases, such as Alzheimer's disease (AD), and Parkinson's disease (PD). Among phytochemicals, sulforaphane (SFN) has demonstrated neuroprotective effects in several in vitro and in vivo studies. In particular, evidence suggests that beneficial effects of SFN could be mainly ascribed to its peculiar ability to activate the Nrf2/ARE pathway. Therefore, SFN appears to be a promising compound with neuroprotective properties that may play an important role in preventing neurode-generation. This contribution focuses on Nrf2/ARE-mediated signaling pathways, particularly in relation to neuroprotective effects of sulforaphane.
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Aim To observe the effects of thin recipe of Buyang Huanwu Decoction on angiogenesis and the signal pathway of Nrf2 / HO-1 after cerebral ischemic injury in rats. Methods Totally 120 SD rats were randomly divided into four groups: sham operation group,model group,Buyang Huanwu Decoction group and thin recipe of Buyang Huanwu Decoction group. The focal cerebral ischemia rat model was established by middle cerebral arterial occlusion. Each group was treated with corresponding treatment. Each group was detected after cerebral ischemia for day 1,day 3 and day 7, respectively. Immunohistochemical method was used to detect the plasma levels of factor VIII related antigen( vWF), determination of microvessel density (MVD). The expression of Nrf2,HO-1 gene and pro-tein in brain tissues was detected by Real-Time PCR (RT-PCR) and Western blot. Results ① Compared with sham-operation group, the expression of vWF in the model group was significantly increased on day 3(P< 0. 05). Compared with model group, the expression levels increased differently in each drug group on day 7 (P < 0. 05). ② The expression of Nrf2, HO-1 gene and protein in sham operation group showed a small a-mount of gamma expression. Compared with sham op-eration group at the same time point, the expression of Nrf2 protein was significantly increased on day 3(P <0. 01). Compared with model group at the same time point, the Nrf2mRNA and protein expression was up-regulated in each drug group. The Nrf2mRNA on day 1,the Nrf2 protein on day 1 and day 7 were significant-ly increased( P < 0. 01). Compared with sham opera-tion group at the same time point, the expression of HO-1mRNA and protein in the model group was signif-icantly increased on day 7(P < 0. 05). Compared with model group at the same time point, the HO-1mRNA on day 3, the HO-1 protein on day 3 and day 7 in each drug group were significantly increased (P < 0. 05,P <0. 01). Conclusions The thin recipe of Buyang Hua-nwu Decoction promotes brain angiogenesis after ische-mia. The effect may be related wih the expression of Nrf2 / HO-1 signal pathway.
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Accumulating evidence strongly supports the involvement of oxidative stress in the etiology of Parkinson's disease (PD). ROS derived from the NOX2-containing NADPH oxidase play important role in the pathogenesis of PD. Our intent was to review the research history about the role of NOX2-containing NADPH oxidase in PD. Neuroprotective effect of heme oxygenase-1 (HO-1) in protecting neurons against PD-related neurotoxin-induced oxidative stress dependant injury, and a major emphasis has been on the relationship between HO-1 and NOX2-containing NADPH oxidase. Specific activation of HO-1 gene expression by pharmacological modulation may represent a novel target for therapeutic treatment of PD through inhibiting NOX/ROS. A new target for PD therapy through inhibiting NOX/ROS, thereby modulating HO-1/NOX2 axis is highlighted.
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PURPOSE: Experimental autoimmune uveoretinitis (EAU) is an animal model of posterior uveitis and heme oxygenase-1 (HO-1) is a well-known anti-oxidant factor. However, there is no report a protective role of HO-1 on EAU in vivo. To verify that HO-1 is induced in EAU by interphotoreceptor retinoid-binding protein (IRBP), that an HO-1 inducers ameliorates the associated inflammation, and that an HO-1 inhibitor exacerbates this inflammation. METHODS: Forty four Lewis rats were given either 40 mol/kg hemin or 40 mol/kg SnPP (tin protoporphyrin IX) by intraperitoneal injection and twenty two uveitis control rats were injected with 0.5 mL of saline once daily 5-20 days after IRBP immunization inducing EAU. Three normal control rats were used for Western blotting and ELISA assay of HO-1. The clinical uveitis signs of inflammation were scored in the three groups from 0 to 4 on alternate three days. To confirm the clinical results, histological and immunohistochemical stain of HO-1 were performed on the day of peak inflammation and Western blotting and ELISA assay of HO-1 were performed on 6th, 12th and 18th day after IRBP immunization. RESULTS: Hemin, an inducer of HO-1, ameliorated the clinical signs of EAU. In contrast, SnPP-treated rats show that the severity of the clinical sign were exacerbated at the peak period of the disease. These results are roughly compatible with histological, immunoblotting, and immunohistochemical evaluations and an ELISA assay of HO-1. CONCLUSIONS: We suggest that HO-1 plays an important protective role in EAU.