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1.
Basic & Clinical Medicine ; (12): 69-76, 2024.
Article in Chinese | WPRIM | ID: wpr-1018574

ABSTRACT

Objective To investigate sodium hydrogen sulfide(NaHS)with function of regulating glutathione(GSH)synthesis to reduce reactive oxygen species(ROS)production in type 2 diabetic cardiomyopathy(DCM).Methods Mouse cardiomyocyte cell line HL-1 was incubated with high concentration of glucose(HG:40 mmol/L)and palmitate(Pal:500 μmol/L)as a cell model of type 2 DCM.HL-1 cells were incubated with NaHS(100 μmol/L),DL-propargylglycin(PPG,1 mmol/L)and N-acetyl-l-cysteine(NAC,5 mmol/L),respectively for 72 hours.The expression of cystathionine-γ-lyase(CSE)and the key enzymes of glutathione production was tested by Western blot.Dihydroethidium(DHE)and dichlorofluoromethane(DCFH)were used to detect the content of ROS in HL-1 cells.Cell viability was detected by CCK8 kit.The content of total GSH was detected.The interaction between muscle specific ring finger protein 1(Murf1)and nuclear factor erythroid-derived 2-related factor 2(Nrf2)and Nrf2 ubiquitylation was determined by co-immunoprecipitation(co-IP).Results Compared with control group,the expression level of CSE,solute carrier family 7 members 11(SLC7A11),glutamate cysteine ligase C(GCLC),glutamate cysteine ligase M(GCLM)and glutathione synthetase(GSS)in HL-1 cells treated incubated with high glucose and palmitate was decreased,however,NaHS was found to restore it.NaHS reduced the content of ROS in HL-1 cells treated with high glucose and palmitate.The interaction between murf1 and Nrf2 was confirmed by co-immunoprecipitation(Co-IP).Compared with NaHS group,the ubiquitylation level of Nrf2 was enhanced in high glucose and palmitate group.Conclusions Sodium hydrosulfide may reduce the ubiquitylation level of Nrf2 and promote the expression of key enzymes of GSH synthesis.

2.
Article in Chinese | WPRIM | ID: wpr-1021366

ABSTRACT

BACKGROUND:Abnormal extracellular matrix accumulation and excessive proliferation of fibroblasts are the main manifestations of pathological scars.Excessive proliferation of fibroblasts leads to the production of large amounts of collagen-based extracellular matrix.Therefore,to investigate the role of fibroblast fibrosis in the formation of pathological scar will provide a new idea for revealing the mechanism of pathological scar and biological therapy. OBJECTIVE:To investigate the effect of RAS-selective lethal small molecule 3(RSL3)on the fibrosis of human pathological scar fibroblasts. METHODS:Then cases of pathological scar tissue and normal skin tissue samples from the same individuals,provided by the Department of Burn Plastic Surgery,General Hospital of Ningxia Medical University,were collected.Fibroblasts of human pathological scar and human normal skin were extracted and used in the following experiments.The general condition of the pathological scar tissue and the normal skin tissue was detected by hematoxylin-eosin staining.The appearance of fibroblasts from pathological scar and normal skin were observed by inverted microscope.The fibroblasts were verified by immunofluorescence assay.The cells were treated with different concentrations of RSL3(1,3,5,7,9,11,13 μmol/L).The inhibitory concentration of RSL3 on fibroblasts was detected by cell counting kit-8.Control group(without treatment)and RSL3 intervention group(treated with 7 μmol/L RSL3 for 24 hours)were set up.The mRNA and protein expressions of glutathione peroxidase 4,type Ⅰ collagen,type Ⅲ collagen and α-smooth muscle actin were detected by Qrt-PCR and western blot,respectively.Level of malondialdehyde in cells was detected.The residual scratch area was measured by cell scratch test after 24 hours to calculate the percentage of residual scratch area. RESULTS AND CONCLUSION:The expression of glutathione peroxidase 4 in the pathological scar group was higher than that in the normal skin group(Mrna:t=3.252,P<0.01;protein:t=5.075,P<0.01).The expression of glutathione peroxidase 4 in the pathological scar fibroblast group was higher than that in the normal skin fibroblast group(Mrna:t=10.32,P<0.01;protein:t=26.22,P<0.01).Compared with the control group,the expression of glutathione peroxidase 4 was decreased(Mrna:t=2.798,P<0.05;protein:t=4.643,P<0.01),the content of malondialdehyde was increased(t=2.917,P<0.05),the expression of type Ⅰ collagen(Mrna:t=15.84,P<0.01;protein:t=4.610,P<0.01),type Ⅲ collagen(Mrna:t=28.86,P<0.01;protein:t=7.713,P<0.01)and α-smooth muscle actin(Mrna:t=2.671,P<0.05;protein:t=7.417,P<0.01)were decreased in the RSL3 intervention group.Compared with the control group,the migration ability was weakened in the RSL3 intervention group(t=14.06,P<0.01).To conclude,RSL3 can inhibit the expression of glutathione peroxidase 4 and then inhibit the ability of fibrosis and migration of pathological scar fibroblasts.

3.
Article in Chinese | WPRIM | ID: wpr-1021588

ABSTRACT

BACKGROUND:With the aging of the global population,the incidence rate of osteoporosis is also increasing.It is very important to further understand its pathogenesis and propose new therapeutic targets.Recent studies have shown that ferroptosis is closely related to the pathogenesis of some bone diseases,such as inflammatory arthritis,osteoporosis and osteoarthritis. OBJECTIVE:To summarize the previous studies on the mechanism of ferroptosis in osteoporosis,so as to provide new therapeutic ideas and potential therapeutic targets for osteoporosis. METHODS:The first author used the computer to search the documents published from 2000 to 2022 in CNKI,WanFang,VIP,PubMed and Web of Science with the key words of"ferroptosis,osteoporosis,osteoblasts,osteoclasts,iron chelators,reactive oxygen species,nuclear factor erythroid 2-related factor 2,heme oxygenase-1,glutathione peroxidase 4,review"in Chinese and English.A total of 70 articles were finally included according to the inclusion criteria. RESULTS AND CONCLUSION:Ferroptosis is significantly different from necrosis,apoptosis and autophagy.In terms of cell morphology and function,it does not have the morphological characteristics of typical necrosis,nor does it have the characteristics of traditional apoptosis,such as cell contraction,chromatin condensation,the formation of apoptotic bodies and the disintegration of cytoskeleton.Contrary to autophagy,ferroptosis does not form a classical closed bilayer membrane structure(autophagic vacuole).Morphologically,ferroptosis is mainly manifested by obvious contraction of mitochondria,increased membrane density,and reduction or disappearance of mitochondrial cristae,which are different from other cell death modes.Iron overload can destroy bone homeostasis by significantly inhibiting osteogenic differentiation and stimulating osteoclast formation,leading to osteoporosis.Iron overload interferes with the differentiation of stem cells to osteoblasts,leading to a weakened osteoblast function and further imbalance of bone metabolism in the body,which eventually leads to osteoporosis.Stimulated by iron overload,osteoclast bone resorption is enhanced and bone loss exceeds new bone formation.Iron chelators have been proved to have osteoprotective effects by inhibiting osteoclast activity and stimulating osteogenic differentiation of osteoblasts.Its potential mechanism is related to inhibiting osteoclast differentiation and promoting osteoblast differentiation.Antioxidants can prevent reactive oxygen species production and inhibit bone absorption,thus improving bone metabolism and effectively preventing osteoporosis.

4.
Article in Chinese | WPRIM | ID: wpr-1021885

ABSTRACT

BACKGROUND:It is of great significance to find new diagnostic markers of the disease and molecular targets for the treatment of the disease and the alleviation of organ injury.Ferroptosis is a newly discovered form of cell death.Overactivation of ferroptosis in animal models of sepsis is associated with the activation of inflammatory response and the injury of the liver,heart,kidney and other important organs,but the relationship between ferroptosis and bloodstream infection is not very clear. OBJECTIVE:To study the changes and biological significance of ferroptosis in a mouse model of blood stream infection induced by different bacteria. METHODS:Blood stream infection models induced by gram negative bacteria Escherichia coli,Klebsiella pneumoniae and gram positive bacteria Staphylococcus aureus and Enterococcus faecalis were established in SPF-grade ICR male mice,with 42 mice in each group.The mRNA expression levels of ferroptosis marker genes transferrin receptor 1 and glutathione peroxidase 4 in the liver,myocardium and kidney were detected at 0.5,1,3,6,12,24 and 48 hours after modeling.Another 18 SPF-grade ICR male mice were selected and randomly divided into dimethyl sulfoxide(DMSO)control group,DMSO+Klebsiella pneumoniae group,and Ferrostatin-1+Klebsiella pneumoniae group,with 6 mice in each group.In the latter two groups,animal models of Klebsiella pneumoniae bloodstream infection were established by tail vein injection of Klebsiella pneumoniae suspension,and 5 mg/kg Ferrostatin-1 and an equal dose of DMSO were given intraperitoneally 1 hour prior to the modeling of bloodstream infection,respectively.Serum levels of alanine aminotransferase,aspartate aminotransferase,blood creatinine,blood urea nitrogen,phosphocreatine kinase isoenzyme,lactate dehydrogenase,and mRNA expression levels of ferroptosis marker genes in various tissues were assayed at 6 hours after modeling. RESULTS AND CONCLUSION:After bloodstream infection modeling,the mRNA expression levels of transferrin receptor 1 in the liver,myocardium and kidney of bloodstream infection mice with different bacteria increased first and then decreased;and the mRNA expression level of glutathione peroxidase 4 decreased first,then increased,and reached the peak at 6 hours after modeling.The changes in transferrin receptor 1 and glutathione peroxidase 4 mRNA levels in bloodstream infection mice induced by gram-negative bacteria were more significant than those in blood stream infection mice induced by gram-positive bacteria,especially in bloodstream infection mice induced by Klebsiella pneumoniae.At 6 hours after bloodstream infection induced by Klebsiella pneumoniae,the levels of alanine aminotransferase,aspartate aminotransferase,serum creatinine,blood urea nitrogen,creatine phosphate kinase isoenzyme,lactate dehydrogenase in mice were significantly increased.Before modeling,Ferrostatin-1 intervention significantly reduced the levels of alanine aminotransferase,aspartate aminotransferase,serum creatinine,blood urea nitrogen,creatine phosphate kinase isoenzyme,and lactate dehydrogenase.All these findings indicate that the activation of ferroptosis in bloodstream infection mice induced by different bacteria is obvious,and the activation of ferroptosis in bloodstream infection mice induced by gram-negative bacteria is more obvious.Inhibition of iron death significantly attenuates liver,myocardial,and kidney injury in the mouse model of bloodstream infection induced by Klebsiella pneumoniae.

5.
Article in Chinese | WPRIM | ID: wpr-1032235

ABSTRACT

Objective @#To evaluate the effect of melatonin on nocturnal exacerbation of neuropathic pain and to ex plore its mechanism through the specific silencing information regulator 1 ( SIRT1)-brain and muscle ARNT-like protein 1 ( BMAL1 ) pathway .@*Methods @# 96 SPF grade male C57/B6 mice were randomly divided into three groups : the sham operation (S) group , the neuropathic pain model (NP) group and the NP model + melatonin treatment (10 mg/kg) ( NP + M) group; preoperative experimental mice were placed in the environment of the specified light pattern; the environment of alternating 12 h light and 12 h darkness was used for at least two weeks , and natural time was converted into the time of the award (ZT) , and the starting point of the light was ZT0; only the sciatic nerve was isolated in the S group , and the mouse NP model was prepared using chronic constriction injury (CCI) of the sciatic nerve in the NP group and the NP + M group , and the NP + M group was inj ected with me latonin after the operation; the expression levels of SIRT1, BMAL1, and glutathione peroxidase 1 (Gpx1) were de tected in the spinal cord at each time point at 14 d postoperatively by Western blot. Postoperative co-staining of SIRT1 in the dorsal horn of the spinal cord with the spinal cord neuronal marker neuron specific nuclear protein (NeuN) , the microglial cell activation marker ion calcium binding adapter molecule 1 (iba-1) , and the astrocyte marker glial fibrillary acidic protein ( GFAP) was carried out by immunofluorescence and iba-1 was detected at each time point to determine the activation status of microglia.@*Results @#SIRT1 , BMAL1 and Gpx1 decreased in NP group mice at 14 d ZT22 postoperatively compared to ZT10 time point in NP group ( P < 0.05) ; SIRT1 and BMAL1 were elevated in NP + M group at ZT14 time point compared to ZT14 time point in NP group (P < 0.05) , whereas Gpx1 was elevated at ZT18 time point (P < 0.05) . SIRT1 was co expressed in the dorsal horn of the spinal cord and in microglia. C ompared with ZT10 time point , microglia expression decreased in NP group mice at ZT22 time point 14 d after surgery (P < 0.05) ; compared with ZT10 time point , there was no statistically significant difference in microglia expression in NP + M group mice at ZT22 time point 14 d after surgery .@*Conclusion@#Melatonin attenuates nocturnal exacerbation of neuropathic pain by a mechanism that may be related to activation of microglia SIRT1-BMAL1 pathway protein expression .

6.
Article in Chinese | WPRIM | ID: wpr-1036225

ABSTRACT

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.

7.
Journal of Clinical Hepatology ; (12): 502-508, 2024.
Article in Chinese | WPRIM | ID: wpr-1013128

ABSTRACT

ObjectiveTo investigate the role and mechanism of action of Yinchenhao Decoction in inhibiting ferroptosis of hepatocytes in mice with autoimmune hepatitis. MethodsA total of 18 specific pathogen-free female C57BL/6 mice were selected and divided into normal group, model group, and treatment group using a random number table, with 6 mice in each group. The mice in the model group and the treatment group were injected with concanavalin A (Con A) via the caudal vein to establish a mouse model of autoimmune hepatitis, and those in the normal group were injected with normal saline. The mice in the treatment group were given prophylactic treatment with Yinchenhao Decoction (4.68 g crude drug/kg) by gavage at 14 days before modeling, and Con A was injected after the last gavage. The levels of alanine aminotransferase (ALT), aspartate aminotransferase (AST), interferon gamma (IFN-γ), tumor necrosis factor-α (TNF-α), iron ion, glutathione (GSH), reactive oxygen species (ROS), adenosine triphosphate (ATP), and malondialdehyde (MDA) were measured; liver index and spleen index were calculated; the expression levels of GPX4 and SLC7A11 were measured; liver histopathological changes were compared between groups. A one-way analysis of variance was used for comparison of normally distributed continuous data between three groups, and the least significant difference t-test was used for further comparison between two groups. ResultsCompared with the normal group, the model group had significant increases in liver index, spleen index, ALT, AST, IFN-γ, TNF-α, iron ion, ROS and MDA (all P<0.05) and significant reductions in the content of GSH and ATP and the protein expression levels of GPX4 and SLC7A11 (all P<0.05). Compared with the model group, the treatment group had significant reductions in liver index, spleen index, ALT, AST, IFN-γ, TNF-α, iron ion, ROS and MDA (all P<0.05) and significant increases in the content of GSH and ATP and the protein expression levels of GPX4 and SLC7A11 (all P<0.05). HE staining showed that compared with the normal group, the model group showed massive hepatocyte degeneration and necrosis and inflammatory cell aggregation at the portal area, and compared with the model group, the treatment group had alleviation of liver necrosis and inflammatory infiltration. ConclusionLiver injury induced by Con A may be associated with ferroptosis. Yinchenhao Decoction can increase the protein expression levels of SLC7A11 and GPX4 protein and thus inhibit ferroptosis of hepatocytes induced by Con A.

8.
Article in Chinese | WPRIM | ID: wpr-1006550

ABSTRACT

ObjectiveTo observe the effect of Banxia Xiexintang (BXT) on the proliferation of human gastric cancer HGC-27, MKN-45, and AGS cells and its mechanism. MethodCell counting kit-8 (CCK-8) was used to detect the effects of different concentrations of BXT-containing serum (5%, 10%, and 20%) on the proliferation of HGC-27, MKN-45, and AGS cells. A mitochondrial membrane potential probe (TMRE) was used to detect the expression of mitochondrial membrane potential in cells. A kit was used to detect iron ion (Fe2+) content, lipid peroxide (LPO), and superoxide dismutase (SOD) activity. Western blot was used to detect the protein expression levels of glycogen synthase3β (GSK3β), phosphorylated GSK3β (p-GSK3β), nuclear factor E2 related factor 2 (Nrf2), and glutathione peroxidase 4 (GPX4). The real-time fluorescence quantitative polymerase chain reaction (Real-time PCR) was used to detect the mRNA expression of member 11 of the cystine/glutamic acid reverse transporter solute vector family 7 (SLC7A11), member 2 of the heavy chain solute vector family 3 (SLC3A2), transferrin receptor 3 (TFRC), and tumor protein (TP)53. ResultCCK-8 results showed that BXT and capecitabine could significantly reduce the survival rate of three kinds of gastric cancer cells after treatment with drug-containing serum for 24 h (P<0.01). After 48 h of intervention with drug-containing serum, the survival rate of three kinds of gastric cancer cells was significantly decreased in both the capecitabine group and the BXT group compared with the blank group. The BXT group was dose-dependent, with 20% BXT having the most significant effect (P<0.01). In terms of biochemical indicators of ferroptosis, compared with the blank group, BXT and capecitabine significantly decreased the expression of mitochondrial membrane potential (P<0.01) and SOD activity (P<0.01) and significantly increased the contents of LPO and Fe2+ (P<0.01), so as to improve the sensitivity of gastric cancer cells to ferroptosis. In terms of the Nrf2/GPX4 pathway, compared with the blank group, the BXT group could reduce the protein expressions of p-GSK3β, Nrf2, and GPX4 (P<0.01) in gastric cancer cells and increase mRNA expressions of SLC7A11 and SLC3A2 (P<0.05). It could also increase the protein expression of GSK3β (P<0.01) and mRNA expression of TP53 and TFRC (P<0.05, P<0.01) in gastric cancer cells. Inhibition of the Nrf2/GPX4 pathway induces ferroptosis in gastric cancer cells. Compared with the capecitabine group, the 20% BXT group showed a more obvious effect. ConclusionBanxia Xiexintang can induce ferroptosis in gastric cancer cells HGC-27, MKN-45, and AGS by inhibiting the Nrf2/GPX4 pathway.

9.
Article in Chinese | WPRIM | ID: wpr-1039625

ABSTRACT

ObjectiveTo investigate the effect of Zuoguiwan on the ovarian function in the rat model of cyclophosphamide-induced premature ovarian failure (POF) based on the changes of ferroptosis pathway. MethodForty SD rats were randomized into blank, model, and low- and high-dose (2, 8 g·kg-1, respectively) Zuoguiwan groups, with 10 rats in each group. The rats in the other groups except the normal group were intraperitoneally injected with CTX at a dose of 50 mg·kg-1 on the first day and 8 mg·kg-1 from the second day to the fifteenth day for the modeling of POF. After modeling, the rats were administrated with corresponding drugs or normal saline by gavage for four weeks. Hematoxylin-eosin staining was performed to observe the pathological changes in the ovarian tissue. The mitochondria of the ovarian tissue was observed by electron microscopy. The serum levels of follicle-stimulating hormone (FSH), estradiol (E2), luteinizing hormone (LH), anti-Mullerian hormone (AMH), malondialdehyde (MDA), catalase (CAT), superoxide dismutase (SOD), and iron ion were measured by biochemical methods and enzyme-linked immunosorbent assay. Western blot was employed to determine the protein levels of glutathione peroxidase 4 (GPX4), solute carrier family 7 member 11 (SLC7A11), ferritin heavy chain (FTH1), and acyl-CoA synthetase long chain family member 4 (ACSL4). ResultCompared with the blank group, the model group showcased significantly increased atretic follicles, atrophied, fragmented, and vacuolated mitochondria, and reduced, loose, and disordered cristae in mitochondria. Compared with the model group, high-dose Zuoguiwan increased mature follicles, the volume of mitochondria in the ovary, alleviated the vacuolation, and improved the number and arrangement of mitochondrial cristae. Compared with the blank group, the modeling elevated the levels of iron, MDA, FSH, and LH, up-regulated the expression of GPX4, SLC7A11, and FTH1 (P<0.05, P<0.01), decreased the activities of SOD and CAT, lowered the levels of E2 and AMH, and down-regulated the expression of ACSL4 (P<0.05, P<0.01). Compared with the model group, drug interventions lowered the levels of iron, MDA, FSH, and LH, down-regulated the expression of GPX4, SLC7A11, and FTH1 (P<0.05, P<0.01), increased the activity of CAT, elevated the levels of E2 and AMH, and up-regulated the expression of ACSL4 (P<0.05, P<0.01). ConclusionZuoguiwan may inhibit the occurrence of ferroptosis by regulating the SLC7A11/GPX4 axis, thereby improving the ovarian function of POF rats.

10.
Herald of Medicine ; (12): 526-534, 2024.
Article in Chinese | WPRIM | ID: wpr-1023744

ABSTRACT

Objective To explore the effect of Shuanglu Tongnao Formula on neuronal ferroptosis in ischemic stroke rats and its regulatory mechanism on the silent information regulator 2 homolog 1(SIRT1)/nuclear factor erythroid 2-related fac-tor 2(Nrf2)/glutathione peroxidase 4(GPx4)signaling pathways.Methods Twenty rats were selected as sham operation group by the random number table method,and the remaining seventy rats were made ischemic stroke rat models by the middle cerebral artery occlusion method.The rats that had been successfully modeled were randomly divided into the model control group,Shuanglu Tongnao formula group,Shuanglu Tongnao formula+SIRT1 inhibitor group(Shuanglu Tongnao formula+EX527 group),with 20 rats in each group.After 14 days,the rats were scored for neurological injury;TTC staining was applied to detect the area of cerebral infarction in rats;HE staining was applied to detect pathological changes in rat brain tissue;Nissl staining was applied to detect the number of neurons in rat brain tissue;the kit was applied to detect the levels of ferri ion(Fe2+),superoxide dismutase(SOD),glutathione(GSH),and malonaldehyde(MDA)in rat brain tissue;immunohistochemistry was applied to de-tect the positive expression of acyl-CoA synthetase long-chain family member 4(ACSL4),transferrin receptor(TFR),and ferritin heavy polypeptide 1(FTH1)proteins in rat brain tissue;Western blotting method was applied to detect the expression of SIRT1,Nrf2,GPx4,and cystine/glutamate antiporter solute carrier family 7 member 11(SLC7A11)proteins in rat brain tissue.Results Compared with the sham operation group,the neurological deficit score,cerebral infarction area,the contents of Fe2+and MDA,and the protein expressions of ACSL4 and TFR in model control group were increased(P<0.05);the number of neurons,the con-tents of SOD and GSH,the protein expression of FTH1,SIRT1,Nrf2,GPx4,and SLC7A11 were all reduced(P<0.05).Compared with the model control group,the neurological deficit score,cerebral infarction area,the contents of Fe2+and MDA,and the protein expression of ACSL4 and TFR in the Shuanglu Tongnao formula group were reduced(P<0.05),and the number of neurons,the contents of SOD and GSH,the protein expressions of FTH1,SIRT1,Nrf2,GPx4,and SLC7A11 are all increased(P<0.05).The results of the SIRT1 inhibitor supplementation experiment showed that the SIRT1 inhibitor reversed the inhibitory effect of Shuan-glu Tongnao formula on neuronal ferroptosis,while also inhibited the expression of Nrf2 and GPx4(P<0.05).Conclusion The Shuanglu Tongnao formula may inhibit neuronal ferroptosis in ischemic stroke rats by activating the SIRT1/Nrf2/GPx4 signa-ling pathway.

11.
Article in Chinese | WPRIM | ID: wpr-1025342

ABSTRACT

Objective:To observe and verify the changes of transcriptome in hyperoxia-induced acute lung injury (HALI), and to further clarify the changes of pathways in HALI.Methods:Twelve healthy male C57BL/6J mice were randomly divided into normoxia group and HALI group according to the random number table, with 6 mice in each group. The mice in the normoxia group were fed normally in the room, and the mice in the HALI group was exposed to 95% oxygen to reproduce the HALI animal model. After 72 hours of hyperoxia exposure, the lung tissues were taken for transcriptome sequencing, and then Kyoto Encyclopedia of Genes and Genomes database (KEGG) pathway enrichment analysis was performed. The pathological changes of lung tissue were observed under light microscope after hematoxylin-eosin (HE) staining. Real-time fluorescence quantitative reverse transcription-polymerase chain reaction (RT-PCR) and Western blotting were used to verify the key molecules in the signal pathways closely related to HALI identified by transcriptomics analysis.Results:Transcriptomic analysis showed that hyperoxia induced 537 differentially expressed genes in lung tissue of mice as compared with the normoxia group including 239 up-regulated genes and 298 down-regulated genes. Further KEGG pathway enrichment analysis identified 20 most significantly enriched pathway entries, and the top three pathways were ferroptosis signaling pathway, p53 signaling pathway and glutathione (GSH) metabolism signaling pathway. The related genes in the ferroptosis signaling pathway included the up-regulated gene heme oxygenase-1 (HO-1) and the down-regulated gene solute carrier family 7 member 11 (SLC7A11). The related genes in the p53 signaling pathway included the up-regulated gene tumor suppressor gene p53 and the down-regulated gene murine double minute 2 (MDM2). The related gene in the GSH metabolic signaling pathway was up-regulated gene glutaredoxin 1 (Grx1). The light microscope showed that the pulmonary alveolar structure of the normoxia group was normal. In the HALI group, the pulmonary alveolar septum widened and thickened, and the alveolar cavity shrank or disappeared. RT-RCR and Western blotting confirmed that compared with the normoxia group, the mRNA and protein expressions of HO-1 and p53 in lung tissue of the HALI group were significantly increased [HO-1 mRNA (2 -ΔΔCt): 2.16±0.17 vs. 1.00±0.00, HO-1 protein (HO-1/β-actin): 1.05±0.01 vs. 0.79±0.01, p53 mRNA (2 -ΔΔCt): 2.52±0.13 vs. 1.00±0.00, p53 protein (p53/β-actin): 1.12±0.02 vs. 0.58±0.03, all P < 0.05], and the mRNA and protein expressions of Grx1, MDM2, SLC7A11 were significantly decreased [Grx1 mRNA (2 -ΔΔCt): 0.53±0.05 vs. 1.00±0.00, Grx1 protein (Grx1/β-actin): 0.54±0.03 vs. 0.93±0.01, MDM2 mRNA (2 -ΔΔCt): 0.48±0.03 vs. 1.00±0.00, MDM2 protein (MDM2/β-actin): 0.57±0.02 vs. 1.05±0.01, SLC7A11 mRNA (2 -ΔΔCt): 0.50±0.06 vs. 1.00±0.00, SLC7A11 protein (SLC7A11/β-actin): 0.72±0.03 vs. 0.98±0.01, all P < 0.05]. Conclusions:HALI is closely related to ferroptosis, p53 and GSH metabolism signaling pathways. Targeting the key targets in ferroptosis, p53 and GSH metabolism signaling pathways may be an important strategy for the prevention and treatment of HALI.

12.
Clinics ; Clinics;79: 100359, 2024.
Article in English | LILACS-Express | LILACS | ID: biblio-1564364

ABSTRACT

Abstract Objective The aim of this study was to evaluate the GSH effect on functional and histological recovery after experimental spinal cord injury in rats. Methods Forty Wistar rats were subjected to spinal cord injury through the Multicenter Animal Spinal Cord Injury Study (MASCIS) Impactor system. The rats were sorted and divided into four groups, as follows: Group 1 ‒ Laminectomy and spinal cord injury; Group 2 ‒ Laminectomy, spinal cord injury and Saline Solution (SS) 0.9%; Group 3 ‒ Laminectomy, spinal cord injury, and GSH; and Group 4 ‒ lLaminectomy without spinal cord injury. GSH and SS were administered intraperitoneally. Groups 1 and 4 received no intervention. Results The rats were evaluated for locomotor function recovery at seven different times by the Basso, Beattie, and Bresnahan (BBB) scale on days 2, 7, 14, 21, 28, 35, and 42 after the spinal cord injury. On day 42, the rats were sacrificed to analyze the histological findings of the injured spinal cord. In the group submitted to GSH, our experimental study revealed better functional scores on the BBB scale, horizontal ladder scale, and cranial and caudal axon count. The differences found were statistically significant in BBB scores and axonal count analysis. Conclusion This study demonstrated that using glutathione in experimental spinal trauma can lead to better functional recovery and improved axonal regeneration rate in Wistar rats submitted to experimental spinal cord injury.

13.
Article | IMSEAR | ID: sea-218094

ABSTRACT

Background: Hypertension is consistently related to the development of ischemic heart disease, heart failure, stroke, and chronic kidney disease. Oxidative stress has been associated with mechanisms of hypertension which could be nullified by antioxidants such as Vitamin C and Vitamin E. Aim and Objectives: The objectives of the study are as follows: (i) To estimate the impact of antioxidant therapy on antioxidant capacity in hypertensive patients; (ii) to measure serum levels of glutathione, glutathione peroxidase (GPx), glutathione reductase (GR), and superoxide dismutase (SOD) in hypertensive patients before and after giving them antioxidant therapy for 45 days. Materials and Methods: Thirty randomly selected hypertensive patients were given Supradyn tablet once a day for 45 days. Ferric reducing ability of plasma (FRAP), SOD, GR, GPx, and reduced Glutathione assays were measured before and after the intervention therapy. Results: Total antioxidant capacity as measured by serum FRAP in hypertensive patients before and after the therapy was increased significantly from 578.8 ± 60.85 to 592.1 ± 59.66 (?mol/L), respectively. The levels of SOD, GPx, GR, and Glutathione in hypertensive patients before giving antioxidant therapy were 1.6 ± 0.49 U/ml, 184.6 ± 17.1 ?mol/L/min, 8.96 ± 1.15 ?mol/L/min, and 8.03 ± 0.96 ?mol/g of Hb, respectively. The same after giving them antioxidant therapy were 1.7 ± 0.46 U/ml, 182.4 ± 15.98 ?mol/L/min, 8.83 ± 1.11 ?mol/L/min, and 7.83 ± 0.94 ?mol/g of Hb, respectively. The levels of GPx, GR, and Glutathione were significantly decreased after giving antioxidant therapy for 45 days while SOD level did not change significantly. Conclusion: Antioxidant therapies for 45 days led to a significant increase in total antioxidant capacity as shown by plasma FRAP levels and a significant decrease in serum levels of enzymatic antioxidants such as GPx, GR and Glutathione in hypertensive patients. However, serum levels of SOD did not show a significant change.

14.
Article in English | LILACS-Express | LILACS | ID: biblio-1535906

ABSTRACT

Introduction: Tuberculosis is an infectious disease that can be prevented and cured, but it is still associated with high morbidity and mortality rates. Disseminated tuberculosis, although rare, can occur in individuals with underlying pathologies that affect the immune system. Currently, there are limited reports on disseminated tuberculosis in individuals with congenital disorders. Clinical case: We present a case of a patient with a history of ß thalassemia who was admitted to the emergency department with symptoms of abdominal pain and constitutional symptoms. The final diagnosis was disseminated tuberculosis. This case is of particular interest due to its atypical presentation, the initial suspicion of malignancy, and the extensive involvement of the disease despite the patient's absence of immunosuppression history. Conclusions: Disseminated tuberculosis in immunocompetent patients is a rare presentation associated with poor outcomes. The history of ß thalassemia may be a risk factor to consider based on the metabolic pathways involved in the pathophysiology of both diseases.


Introducción: la tuberculosis es una enfermedad infecciosa prevenible y curable asociada a una alta morbimortalidad, la presentación de tuberculosis diseminada es poco frecuente y está asociada a patologías que comprometen el sistema inmunitario. En la actualidad hay pocos informes sobre tuberculosis diseminada y trastornos congénitos subyacentes. Caso clínico: paciente con antecedente de talasemia ß que ingresó al servicio de urgencias por dolor abdominal y síntomas constitucionales con diagnóstico final de tuberculosis diseminada. Es un caso de especial interés debido a la presentación atípica, la sospecha diagnóstica inicial de malignidad y el amplio compromiso de la enfermedad a pesar de que el paciente no tenía antecedentes de inmunosupresión. Conclusiones: la tuberculosis diseminada en el paciente inmunocompetente es una presentación poco frecuente asociada a desenlaces adversos. El antecedente de talasemia ß podría ser un factor de riesgo para tener en cuenta con base en las vías metabólicas involucradas en la fisiopatología de ambas enfermedades.

15.
Braz. J. Psychiatry (São Paulo, 1999, Impr.) ; Braz. J. Psychiatry (São Paulo, 1999, Impr.);45(2): 117-126, Mar.-Apr. 2023. tab, graf
Article in English | LILACS-Express | LILACS | ID: biblio-1439560

ABSTRACT

Objective: The clinical trajectories of patients with psychotic disorders have divergent outcomes, which may result in part from glutathione (GSH)-related high-risk genotypes. We aimed to determine pharmacokinetics of clozapine, GSH levels, GSH peroxidase (GPx) activity, gene variants involved in the synthesis and metabolism of GSH, and their association with psychotic disorders in Mexican patients on clozapine monotherapy and controls. Methods: The sample included 75 patients with psychotic disorders on clozapine therapy and 40 paired healthy controls. Plasma clozapine/N-desmethylclozapine, GSH concentrations, and GPx activity were determined, along with genotyping of GCLC and GSTP1 variants and copy number variations of GSTP1, GSTT1, and GSTM1. Clinical, molecular and biochemical data were analyzed with a logistic regression model. Results: GSH levels were significantly reduced and, conversely, GPx activity was higher among patients than controls. GCLC_GAG-7/9 genotype (OR = 4.3, 95%CI = 1.40-14.31, p = 0.019) and hetero-/homozygous genotypes of GCLC_rs761142 (OR = 6.09, 95%CI = 1.93-22.59, p = 0.003) were found to be risk factors for psychosis. The genetic variants were not related to clozapine/N-desmethylclozapine levels or metabolic ratio. Conclusions: GCLC variants were associated with the oxidative stress profile of patients with psychotic disorders, raising opportunities for intervention to improve their antioxidant defenses. Further studies with larger samples should explore this proposal.

16.
Article | IMSEAR | ID: sea-217964

ABSTRACT

Background: Diabetes, hypertension, obesity, and dyslipidemia, all are the risk factors of metabolic syndrome (MS). Various studies have shown that each risk factor is associated with increased inflammation. hsCRP is a non-specific, sensitive inflammatory marker that is raised in various inflammatory conditions. Similarly, glutathione is an antioxidant which binds with ROS produced during inflammation and reduces damage caused by ROS. Aims and Objectives: This study has been planned to find the correlation between oxidative stress and metabolic risk factors in apparently healthy adults. Materials and Methods: We recruited apparently healthy adults (n = 120) and measured waist circumference, blood pressure, lipid profile, Fasting blood sugar, serum GSH, and hsCRP in all the subjects. Seventy-seven subjects were found to have at least one or more metabolic risk factors (Group A) according to NCEP ATP III criteria with waist circumference >90 cm for male and >80 cm for female and 43 were without any metabolic risk factors (Group B). Thereafter, we compared the serum levels of hsCRP and serum GSH with persons having one or more risk factors for MS. Results: In this study, we observed that subjects with metabolic risk factors were having more oxidative stress indicated by increased hsCRP (4783.1 ± 2060.21) and low serum GSH (3.17 ± 0.81) in comparison to controls (1640.5 ± 547.47 and 4.79 ± 0.77, respectively). This increase in hsCRP and decrease in GSH in case group was statistically significant. We also found the higher basal hsCRP levels in control group as per AHA/CDC study. Conclusion: We observed in this study that Indians without any risk factors for MS have relatively higher CRP levels and are at intermediate risk for cardiovascular disease. It was also observed that as the number of metabolic risk factors increases, the levels of hsCRP increases, and serum GSH decreases. This indicates that more risk factors are associated with higher oxidative stress.

17.
Article in English | WPRIM | ID: wpr-971474

ABSTRACT

Ex vivo culture-amplified mesenchymal stem cells (MSCs) have been studied because of their capacity for healing tissue injury. MSC transplantation is a valid approach for promoting the repair of damaged tissues and replacement of lost cells or to safeguard surviving cells, but currently the efficiency of MSC transplantation is constrained by the extensive loss of MSCs during the short post-transplantation period. Hence, strategies to increase the efficacy of MSC treatment are urgently needed. Iron overload, reactive oxygen species deposition, and decreased antioxidant capacity suppress the proliferation and regeneration of MSCs, thereby hastening cell death. Notably, oxidative stress (OS) and deficient antioxidant defense induced by iron overload can result in ferroptosis. Ferroptosis may inhibit cell survival after MSC transplantation, thereby reducing clinical efficacy. In this review, we explore the role of ferroptosis in MSC performance. Given that little research has focused on ferroptosis in transplanted MSCs, further study is urgently needed to enhance the in vivo implantation, function, and duration of MSCs.


Subject(s)
Humans , Antioxidants/metabolism , Ferroptosis , Mesenchymal Stem Cell Transplantation , Mesenchymal Stem Cells , Iron Overload/metabolism
18.
Acta Pharmaceutica Sinica B ; (6): 298-314, 2023.
Article in English | WPRIM | ID: wpr-971689

ABSTRACT

Metastasis accounts for 90% of breast cancer deaths, where the lethality could be attributed to the poor drug accumulation at the metastatic loci. The tolerance to chemotherapy induced by breast cancer stem cells (BCSCs) and their particular redox microenvironment further aggravate the therapeutic dilemma. To be specific, therapy-resistant BCSCs can differentiate into heterogeneous tumor cells constantly, and simultaneously dynamic maintenance of redox homeostasis promote tumor cells to retro-differentiate into stem-like state in response to cytotoxic chemotherapy. Herein, we develop a specifically-designed biomimic platform employing neutrophil membrane as shell to inherit a neutrophil-like tumor-targeting capability, and anchored chemotherapeutic and BCSCs-differentiating reagents with nitroimidazole (NI) to yield two hypoxia-responsive prodrugs, which could be encapsulated into a polymeric nitroimidazole core. The platform can actively target the lung metastasis sites of triple negative breast cancer (TNBC), and release the escorted drugs upon being triggered by the hypoxia microenvironment. During the responsiveness, the differentiating agent could promote transferring BCSCs into non-BCSCs, and simultaneously the nitroimidazole moieties conjugated on the polymer and prodrugs could modulate the tumor microenvironment by depleting nicotinamide adenine dinucleotide phosphate hydrogen (NADPH) and amplifying intracellular oxidative stress to prevent tumor cells retro-differentiation into BCSCs. In combination, the BCSCs differentiation and tumor microenvironment modulation synergistically could enhance the chemotherapeutic cytotoxicity, and remarkably suppress tumor growth and lung metastasis. Hopefully, this work can provide a new insight in to comprehensively treat TNBC and lung metastasis using a versatile platform.

19.
Acta Pharmaceutica Sinica B ; (6): 577-597, 2023.
Article in English | WPRIM | ID: wpr-971714

ABSTRACT

Amyotrophic lateral sclerosis (ALS) is a progressive neurodegenerative disease affecting both upper and lower motor neurons (MNs) with large unmet medical needs. Multiple pathological mechanisms are considered to contribute to the progression of ALS, including neuronal oxidative stress and mitochondrial dysfunction. Honokiol (HNK) has been reported to exert therapeutic effects in several neurologic disease models including ischemia stroke, Alzheimer's disease and Parkinson's disease. Here we found that honokiol also exhibited protective effects in ALS disease models both in vitro and in vivo. Honokiol improved the viability of NSC-34 motor neuron-like cells that expressed the mutant G93A SOD1 proteins (SOD1-G93A cells for short). Mechanistical studies revealed that honokiol alleviated cellular oxidative stress by enhancing glutathione (GSH) synthesis and activating the nuclear factor erythroid 2-related factor 2 (NRF2)-antioxidant response element (ARE) pathway. Also, honokiol improved both mitochondrial function and morphology via fine-tuning mitochondrial dynamics in SOD1-G93A cells. Importantly, honokiol extended the lifespan of the SOD1-G93A transgenic mice and improved the motor function. The improvement of antioxidant capacity and mitochondrial function was further confirmed in the spinal cord and gastrocnemius muscle in mice. Overall, honokiol showed promising preclinical potential as a multiple target drug for ALS treatment.

20.
Chinese Journal of Biotechnology ; (12): 3318-3335, 2023.
Article in Chinese | WPRIM | ID: wpr-1007960

ABSTRACT

ATP is an important cofactor involved in many biocatalytic reactions that require energy input. Polyphosphate kinases (PPK) can provide energy for ATP-consuming reactions due to their cheap and readily available substrate polyphosphate. We selected ChPPK from Cytophaga hutchinsonii for substrate profiling and tolerance analysis. By molecular docking and site-directed mutagenesis, we rationally engineered the dual-substrate channel cavity of polyphosphate kinase to improve the catalytic activity of PPK. Compared with the wild type, the relative enzyme activity of the screened mutant ChPPKK81H-K103V increased by 326.7%. Meanwhile, the double mutation expanded the substrate utilization range and tolerance of ChPPK, and improved its heat and alkali resistance. Subsequently, we coupled the glutathione bifunctional enzyme GshAB and ChPPKK81H-K103V based on this ATP regeneration system, and glutathione was produced by cell-free catalysis upon disruption of cells. This system produced (25.4±1.9) mmol/L glutathione in 6 h upon addition of 5 mmol/L ATP. Compared with the system before mutation, glutathione production was increased by 41.9%. After optimizing the buffer, bacterial mass and feeding time of this system, (45.2±1.8) mmol/L glutathione was produced in 6 h and the conversion rate of the substrate l-cysteine was 90.4%. Increasing the ability of ChPPK enzyme to produce ATP can effectively enhance the conversion rate of substrate and reduce the catalytic cost, achieving high yield, high conversion rate and high economic value for glutathione production by cell-free catalysis. This study provides a green and efficient ATP regeneration system that may further power the ATP-consuming biocatalytic reaction platform.


Subject(s)
Molecular Docking Simulation , Catalysis , Glutathione , Adenosine Triphosphate
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