Effect of octreotide on oxidative stress in the erythrocyte and kidney tissue in adriamycin-induced experimental nephrotic syndrome model / Efeito do octreotide no estresse oxidativo em eritrócitos e no tecido renal no modelo de síndrome nefrótica experimental induzida por adriamicina

Abstract Introduction: Nephrotic syndrome (NS) is one of the reasons of end-stage kidney disease, and elucidating the pathogenesis and offer new treatment options is important. Oxidative stress might trigger pathogenesis systemically or isolated in the kidneys. Octreotide (OCT) has beneficial antioxidant effects. We aimed to investigate the source of oxidative stress and the effect of OCT on experimental NS model. Methods: Twenty-four non-uremic Wistar albino rats were divided into 3 groups. Control group, 2 mL saline intramuscular (im); NS group, adriamycin 5 mg/kg intravenous (iv); NS treatment group, adriamycin 5 mg/kg (iv) and OCT 200 mcg/kg (im) were administered at baseline (Day 0). At the end of 21 days, creatinine and protein levels were measured in 24-hour urine samples. Erythrocyte and renal catalase (CAT) and thiobarbituric acid reactive substance (TBARS) were measured. Renal histology was also evaluated. Results: There was no significant difference among the 3 groups in terms of CAT and TBARS in erythrocytes. Renal CAT level was lowest in NS group, and significantly lower than the control group. In treatment group, CAT level significantly increased compared with NS group. In terms of renal histology, tubular and interstitial evaluations were similar in all groups. Glomerular score was significantly higher in NS group compared with control group and it was significantly decreased in treatment group compared to NS group. Conclusions: Oxidative stress in NS might be due to the decrease in antioxidant protection mechanism in kidney. Octreotide improves antioxidant levels and histology in renal tissue and might be a treatment option.

Resumo Introdução: Síndrome nefrótica (SN) é uma das causas de doença renal em estágio terminal. É importante elucidar a patogênese e oferecer novas opções de tratamento. Estresse oxidativo pode desencadear a patogênese sistemicamente ou isoladamente nos rins. O octreotide (OCT) tem efeitos antioxidantes benéficos. Nosso objetivo foi investigar a fonte de estresse oxidativo e efeito do OCT no modelo experimental de SN. Métodos: Dividimos 24 ratos albinos Wistar não urêmicos em 3 grupos. Grupo controle, 2 mL de solução salina intramuscular (im); grupo SN, adriamicina 5 mg/kg intravenosa (iv); grupo tratamento SN, adriamicina 5 mg/kg (iv) e OCT 200 mcg/kg (im) foram administrados no início do estudo (Dia 0). Aos 21 dias, mediram-se os níveis de creatinina e proteína em amostras de urina de 24 horas. Mediu-se a catalase (CAT) eritrocitária e renal e a substância reativa ao ácido tiobarbitúrico (TBARS). Avaliou-se também histologia renal. Resultados: Não houve diferença significativa entre os três grupos em termos de CAT e TBARS em eritrócitos. O nível de CAT renal foi menor no grupo SN e significativamente menor que no grupo controle. No grupo tratamento, o nível de CAT aumentou significativamente em comparação com o grupo SN. Quanto à histologia renal, as avaliações tubular e intersticial foram semelhantes em todos os grupos. O escore glomerular foi significativamente maior no grupo SN em comparação com o grupo controle e diminuiu significativamente no grupo de tratamento em comparação com o grupo SN. Conclusões: Estresse oxidativo na SN pode ser devido à diminuição do mecanismo de proteção antioxidante nos rins. O octreotide melhora níveis de antioxidantes e histologia do tecido renal e pode ser uma opção de tratamento.

Synergy between isobavachalcone and doxorubicin suppressed the progression of anaplastic thyroid cancer through ferroptosis activation

The objective of this study was to explore the effects and mechanisms of the combination of isobavachalcone (IBC) and doxorubicin (DOX) on the progression of anaplastic thyroid cancer (ATC). Cell viability of 8505C and CAL62 cells was observed by CCK-8 assay. Kits were used to detect the presence of reactive oxygen species (ROS), glutathione (GSH), malondialdehyde (MDA), and cellular iron. Protein expression of solute carrier family 7 member 11 (SLC7A11) and glutathione peroxidase 4 (GPX4) was detected using western blot, and CD31 was detected through immunofluorescence. Tumor xenograft models of 8505C cells were constructed to observe the effect of IBC and DOX on ATC growth in vivo. The co-administration of IBC and DOX exhibited a synergistic effect of suppressing the growth of 8505C and CAL62 cells. The concurrent use of IBC and DOX resulted in elevated iron, ROS, and MDA levels, while reducing GSH levels and protein expression of SLC7A11 and GPX4. However, the Fer-1 ferroptosis inhibitor effectively counteracted this effect. In vitro and in vivo, the inhibitory effect on ATC cell proliferation and tumor growth was significantly enhanced by the combination of IBC and DOX. The combination of IBC and DOX can inhibit the growth of ATC by activating ferroptosis, and might prove to be a potent chemotherapy protocol for addressing ATC.

Research progress on association between macrophages and ischemia-reperfusion injury / 器官移植

Ischemia-reperfusion injury (IRI) is an extremely complicated pathophysiological process, which may occur during the process of myocardial infarction, stroke, organ transplantation and temporary interruption of blood flow during surgery, etc. As key molecules of immune system, macrophages play a vital role in the pathogenesis of IRI. M1 macrophages are pro-inflammatory cells and participate in the elimination of pathogens. M2 macrophages exert anti-inflammatory effect and participate in tissue repair and remodeling and extracellular matrix remodeling. The balance between macrophage phenotypes is of significance for the outcome and treatment of IRI. This article reviewed the role of macrophages in IRI, including the balance between M1/M2 macrophage phenotype, the mechanism of infiltration and recruitment into different ischemic tissues. In addition, the potential therapeutic strategies of targeting macrophages during IRI were also discussed, aiming to provide reference for alleviating IRI and promoting tissue repair.

Down-regulation of XBP1s alleviates the senescence of renal tubular epithelial cells induced by hypoxia/reoxygenation through Sirt3/SOD2/mtROS signaling pathway / 器官移植

Objective To investigate the role and mechanism of spliced X-box binding protein 1 (XBP1s) in the senescence of primary renal tubular epithelial cells induced by hypoxia/reoxygenation (H/R). Methods Primary renal tubular epithelial cells were divided into the normal control group (NC group), H/R group, empty adenovirus negative control group (Ad-shNC group), targeted silencing XBP1s adenovirus group (Ad-shXBP1s group), empty adenovirus+H/R treatment group (Ad-shNC+H/R group) and targeted silencing XBP1s adenovirus+H/R treatment group (Ad-shXBP1s +H/R group), respectively. The expression levels of XBP1s in the NC, H/R, Ad-shNC and Ad-shXBP1s groups were measured. The number of cells stained with β-galactosidase, the expression levels of cell aging markers including p53, p21 and γH2AX, and the levels of reactive oxygen species (ROS), malondialdehyde (MDA) and superoxide dismutase (SOD) were determined in the Ad-shNC, Ad-shNC+H/R and Ad-shXBP1s+H/R groups. Chromatin immunoprecipitation was employed to verify Sirtuin 3 (Sirt3) of XBP1s transcription regulation, and the expression levels of Sirt3 and downstream SOD2 after down-regulation of XBP1s were detected. Mitochondrial reactive oxygen species (mtROS) were detected by flow cytometry. Results Compared with the NC group, the expression level of XBP1s was up-regulated in the H/R group. Compared with the Ad-shNC group, the expression level of XBP1s was down-regulated in the Ad-shXBP1s group (both P<0.001). Compared with the Ad-shNC group, the number of cells stained with β-galactosidase was increased, the expression levels of p53, p21 and γH2AX were up-regulated, the levels of ROS, MDA and mtROS were increased, the SOD activity was decreased, the expression level of Sirt3 was down-regulated, and the ratio of Ac-SOD2/SOD2 was increased in the Ad-shNC+H/R group. Compared with the Ad-shNC+H/R group, the number of cells stained with β-galactosidase was decreased, the expression levels of p53, p21 and γH2AX were down-regulated, the levels of ROS, MDA and mtROS were decreased, the SOD activity was increased, the expression level of Sirt3 was up-regulated and the ratio of Ac-SOD2/SOD2 was decreased in the Ad-shXBP1s+H/R group (all P<0.05). Conclusions Down-regulation of XBP1s may ameliorate the senescence of primary renal tubular epithelial cells induced by H/R, which probably plays a role through the Sirt3/SOD2/mtROS signaling pathway.

Quality Improvement Pathway of Cultivated Chinese Medicinal Materials Based on Physiological Ecology of Plants Under Adversity / 中国实验方剂学杂志

The quality of Chinese medicinal materials is related to the environment， with an optimal quality under adversity. The origin of Chinese medicinal materials has converted from wild collection to cultivation， and a better cultivation environment leads to a decline in their quality. At present， there are few effective methods to improve the quality of Chinese medicinal materials. Plants are bound to produce a large amount of reactive oxygen species （ROS） under adversity， and the quality improvement of Chinese medicinal materials under adversity may be achieved through ROS. This paper described the relationship between plant adversity-ROS-secondary metabolism： ROS can alter the structure of proteins （including enzymes） and regulate enzyme activities， thus affecting secondary metabolism to improve the adaptive capacity of plants. Therefore， ROS is the essential cause of adversity changing secondary metabolism. The cells of plants are omnipotent， and the medicinal parts of plants can independently complete the whole process of secondary metabolism， so regulation of secondary metabolism during the processing of fresh Chinese medicinal materials can significantly improve the quality of Chinese medicinal materials. Exogenous ROS can be used as inducible factors to stimulate medicinal parts， inducing a physiological state of fresh medicinal parts similar to that under adversity， thus enhancing secondary metabolism， and improving the contents of active ingredients in Chinese medicinal materials. In addition， the content and ratio of each ingredient in Chinese medicinal materials are closer to those of wild Chinese medicinal materials. The mechanism of plant adaptation to adversity is the mechanism of the quality formation of Chinese medicinal materials， and the application of ROS as inducible factors can provide a new pathway for the production of high-quality Chinese medicinal materials.

Prevention and Treatment of Alzheimer's Disease by Traditional Chinese Medicine via Regulating ROS： A Review / 中国实验方剂学杂志

The pathological mechanism of Alzheimer's disease （AD） is complex， and there are many hypotheses. The mainstream theory is the amyloid-beta protein （Aβ） and Tau protein phosphorylation. Oxidative stress （OS） is a bridge between other hypotheses and mechanisms and plays a key role in many hypotheses. Therefore， the treatment of OS in AD （ADOS） is beneficial in alleviating disease progression. Reactive oxygen species （ROS） is a kind of antioxidant and a kind of oxidation products， with Aβ and Tau protein interactions， activating microglia and astrocytes， triggering inflammation and mitochondrial dysfunction， leading to the deterioration of the environment in the brain， and accelerating the development of disease. ROS， as a signal messenger inducing OS， is widely involved in the progression of AD and may be a new target for the progression of AD. Traditional Chinese medicine （TCM） monomers and compounds play an increasingly important role in the prevention and treatment of AD. Recent studies have found that the effective prevention and treatment of AD by TCM is closely related to the regulation of ROS. There are many studies on the mechanism of TCM in the treatment of AD via regulating ROS， but there is a lack of systematic review. By analyzing and summarizing the literature in China and abroad in recent years， this paper reviewed the generation and physiology of ROS， the mechanism of action of AD， and the prevention of AD by TCM via regulating ROS through relevant ways， so as to provide references for the research on the regulation of ROS by TCM and provide new targets and new methods for the prevention and treatment of AD.

Effect of Jiegeng Yuanshen Tang on airway inflammation and mucus secretion in allergic asthmatic mice and its mechanism / 吉林大学学报(医学版)

Objective:To discuss the effect of Jiegeng Yuanshen Tang(JGYST)on airway tissue inflammation and mucus secretion in the mice with allergic asthma,and to clarify the related mechanism.Methods:Forty male C57BL/J mice were randomly divided into control group,JGYST group,ovalbumin(OVA)group,and OVA + JGYST group.The mice in OVA group and OVA +JGYST group were sensitized with 50 μg OVA via intraperitoneal injection twice weekly,followed by 20 μg OVA nasal drops daily for 7 d to induce asthma;the mice in OVA +JGYST group were gavaged with 200 μL JGYST 1 h before each OVA challenge,and the administration lasted for 7 d;the mice in control group were given equivalent dose of PBS via intraperitoneal injection,nasal drops,and gavage;the mice in JGYST group were given the same dose of PBS for intraperitoneal and nasal administration and gavaged with the same dose of JGYST.The pathomorphology of lung tissue of the mice in various groups was observed by HE staining and periodic acid-Schiff(PAS)staining,and the inflammation and PAS scores were calculated;flow cytometry method was used to detect the numbers of eosinophils,neutrophils,helper T lymphocyte 1(Th1)cells,helper T lymphocyte 2(Th2)cells,and dendritic cells(DCs),as well as the percentage of mature DCs and level of reactive oxygen species(ROS)in lung tissue of the mice in various groups;real-time fluorescence quantitative PCR(RT-qPCR)method was used to detect the expression levels of interleukin-4(IL-4),interleukin-10(IL-10),and tumor necrosis factor-α(TNF-α)mRNA in lung tissue of the mice in various groups.Results:The HE and PAS staining results showed that the mice in control group had intact airway and alveolar structure,without infiltration of inflammatory cells or mucus secretion;compared with control group,there was a large number of infiltrating inflammatory cells in airway tissue of the mice in OVA group,and the inflammation and PAS scores were increased(P<0.01);compared with OVA group,the infiltration of inflammatory cells in airway tissue of the mice in JGYST group and OVA + JGYST group was decreased,and the inflammation and PAS scores were significantly decreased(P<0.01).The flow cytometry results showed that compared with control group,the numbers of eosinophils,Th2 cells,and DCs in lung tissue of the mice in OVA group were increased(P<0.05 or P<0.01),and the percentage of mature DCs and level of ROS were significantly increased(P<0.01);compared with OVA group,the numbers of eosinophils,Th2 cells,and DCs in lung tissue of the mice in JGYST group and OVA + JGYST group were decreased(P<0.01),and the percentage of mature DCs and level of ROS were significantly decreased(P<0.01).The RT-qPCR results showed that compared with control group,the expression levels of IL-4,IL-10,and TNF-α mRNA in lung tissue of the mice in OVA group were increased(P<0.01);compared with OVA group,the expression levels of IL-4 and TNF-α mRNA in lung tissue of the mice in JGYST group and OVA + JGYST group were decreased(P<0.01),while the expression level of IL-10 mRNA was increased(P<0.01).Conclusion:JGYST can alleviate the airway tissue inflammation and mucus secretion in the mice with allergic asthma,and its mechanism may be related to reducing the number of Th2 cells and DCs,decreasing the ROS level and expression level of proinflammatory cytokine,and increasing the expression level of anti-inflammatory cytokine.

Effects of Tongdu Qishen electroacupuncture on PKC/NADPH oxidative stress pathway in hypothalamus of chronic unpredictable stress depression rats / 国际中医中药杂志

Objective:To observe the antidepressant effect of Tongdu Qishen electroacupuncture method; To explore its mechanism of regulating the oxidative stress pathway of protein kinase C (PKC)/reduced coenzymeⅡ (NADPH) in depression model rats.Methods:Totally 32 SD rats were divided into control group, model group, Tongdu Qishen electroacupuncture group and escitalopram group according to random number table method, with 8 rats in each group. The model of depression was established by chronic unpredictable stress except control group. After the start of modeling, Tongdu Qishen electroacupuncture group was treated with electroacupuncture every day, 15 min/time/day; escitalopram group was given 30 mg/kg intragastric intervention. 1 day before the start of the experiment and the 28th day of the experiment, the growth of body mass was observed, and sugar preference experiment and open field experiment were performed. The protein expression levels of protein kinase C α (PKC α), p47phox, t and RAS related C3 botulinum toxin substrate 1 (Rac1) in hypothalamus were detected by Western blot, and the positive area ratio of NOX2 protein in hypothalamus was detected by immunofluorescence technique; ROS content in hypothalamus was detected using DCFH-DA fluorescent probe technique.Results:Compared with the model group, the Tongdu Qishen electroacupuncture group and the escitalopram group showed the body mass growth ( P<0.01) and sugar preference index increased ( P<0.01), and the moving distance ( P<0.05) and residence time ( P<0.01) in the central area of the open field experiment were longer; the protein expression levels of hypothalamic PKC α, p47phox and Rac1 decreased ( P<0.05 or P<0.01), the positive area ratio of NOX2 protein decreased ( P<0.05), and the level of ROS also decreased significantly ( P<0.01) in Tongdu Qishen electroacupuncture group and escitalopram group. Conclusion:Tongdu Qishen electroacupuncture group can improve the behavior of depressed rats, inhibit the oxidative stress response of PKC/NADPH pathway, and reduce the production of ROS, thereby reducing the brain damage caused by oxidative stress, and improving the symptoms of depression.

Effect of ceria nanoparticles on activity of DSS-induced colitis in mice by eliminating active oxygen species / 上海交通大学学报（医学版）

Objective·To investigate the effect of ceria nanoparticles-polyethylene glycol(CeNP-PEG)on scavenging reactive oxygen species(ROS)and alleviating disease activity in dextran sulphate sodium(DSS)-induced colitis mice.Methods·CeNP was synthesized with the hydrates of cerium acetate,oleamine,and xylene,which was modified with polyethylene glycol-stearyl phosphatidylethanolamine(mPEG-DPSE)to obtain CeNP-PEG.Then CeNP-PEG was purified.The particle size and zeta potential of CeNP-PEG were measured by using transmission electron microscopy(TEM)and dynamic light scattering(DLS).Mouse macrophages(Raw264.7)were cultured in vitro and induced to a pro-inflammatory phenotype(M1 phenotype).M1 macrophages were treated with 0.5 μg/mL and 1.0 μg/mL CeNP-PEG,respectively,and then Western blotting was used to detect the expression changes of the proteins related with nuclear factor-κB(NF-κB)signaling pathway.DSS-induced colitis mice models were constructed,and CeNP-PEG(1.0 mg/mL)was intravenously administrated for 3 times via tail vein during the modeling period.Meanwhile,the body weight,fecal characteristics,and frequency of rectal bleeding in mice were monitored in the normal control group(Normal group),the model group(DSS group),and the CeNP-PEG treatment group.The disease activity index(DAI)was calculated to evaluate the intestinal inflammation.The level of ROS in mouse intestinal tissues was detected by dihydroethidine(DHE)staining and the mRNA expression levels of inflammatory cytokines interferon-γ(Ifn-γ),interleukin-6(Il-6),Il-1β and tumor necrosis factor-α(Tnf-α)were detected by real-time quantitative PCR(RT-qPCR).Results·The hydrated particle size of synthesized CeNP-PEG was(6.96±0.27)nm,and the average zeta potential was(-6.02±1.31)mV.Western blotting results showed that the expression of p-P65 increased in the pro-inflammatory macrophages compared with the control group.The expression of NF-κB inhibitor-α(IκB-α)decreased,and their expressions tended to recover after the intervention of different concentrations of CeNP-PEG.In the DSS-induced colitis models,mice in the CeNP-PEG treatment group lost less weight than those in the DSS group(P= 0.000)and had lower DAI scores(P=0.000).The RT-qPCR results of intestinal tissues showed that the mRNA levels of Ifn-γ,Il-1β,Il-6 and Tnf-α in the DSS group were significantly up-regulated compared with those in the Normal group(P=0.000),and all of them significantly decreased in the CeNP-PEG treatment group.The results of DHE staining showed that the fluorescence intensity of intestinal tissues in the DSS group was significantly enhanced than that in the Normal group,and the fluorescence intensity decreased in the CeNP-PEG treatment group.Conclusion·CeNP-PEG can inhibit the expression of intestinal inflammatory factors and the activation of NF-κB-related inflammatory pathway of pro-inflammatory macrophages,eliminate intestinal ROS,improve the intestinal inflammatory microenvironment,and alleviate the disease activity of DSS-induced colitis in mice.

Mechanism of BMAL1 attenuating H2O2-induced cardiomyocyte injury / 天津医药

Objective To investigate the effect of BMAL1 on H2O2-induced cardiomyocyte injury through NRF2-regulated ROS/NLRP3 inflammasome pathway.Methods H9c2 cells and H9c2 cells with stable over-expressed BMAL1 were cultured and divided into the control group,the H2O2 group,the BMAL1-OE group,the BMAL1-OE+H2O2 group,the BMAL1-OE+ML385 group and the BMAL1-OE+ML385+H2O2 group.All groups were pre-intervened with corresponding inhibitors,and then treated with 0.2 mmol/L H2O2,except for the control group and the BMAL1-OE group.After the intervention,CCK-8 assay was used to measure cell viability,fluorescent probe DCFH-DA was used to measure ROS generation and Western blot assay was used to detect BMAL1,NRF2 and NLRP3 protein expressions.ELISA was used to determine IL-1β release.Results Compared with the control group,the cell viability was decreased,ROS generation was increased,BMAL1 and NRF2 protein expressions were decreased,NLRP3 expression and IL-1β release were increased in the H2O2 group(P＜0.05).Compared with the H2O2 group,the cell viability was increased,ROS generation was decreased,BMAL1-OE and NRF2 protein expressions were increased,NLRP3 expression and IL-1β release were decreased in the BMAL1-OE+H2O2 group(P＜0.05).Compared with the BMAL1-OE+H2O2 group,the cell viability was decreased,ROS generation was increased,NLRP3 expression and IL-1β release were increased in the BMAL1-OE+ML385+H2O2 group(P＜0.05).Conclusion BMAL1 attenuates H2O2-induced H9c2 cardiomyocyte injury,and its mechanism may be related to the regulation of ROS/NLRP3 inflammasome pathway through NRF2.

Molecular mechanisms of Ca2+-induced pyroptosis and adhesion changes of HK-2 cells in the formation of calcium-containing kidney stones / 天津医药

Objective To investigate the possible role and mechanism of activation of pyroptosis classical pathway and alterations in cell adhesion in calcium-containing kidney stones after the action of high concentration of Ca2+ on HK-2 cells.Methods HK-2 cells were cultured in the presence of different concentrations of CaCl2(0,0.1,0.5,1.0,2.0,4.0 and 8.0 g/L)for 24 hours,and cell counting Kit-8(CCK-8)and flow cytometry were used to determine the optimal treatment concentration.Subsequently,the ultrastructure of renal tubular epithelial cells under high Ca2+ condition was observed by transmission electron microscopy after Ca2+ treatment.DCFH-DA staining was used to detect intracellular reactive oxygen species production,and quantitative real-time polymerase chain reaction(qRT-PCR)and Western blot analysis were performed to examine the expression of pyroptosis-related proteins NLRP3,Caspase-1,gasdermin D(GSDMD),adhesive molecules osteopontin(OPN)and CD44 at mRNA and protein levels after high concentration Ca2+ treatment.The expression levels of pyroptosis-related inflammatory factors interleukin(IL)-1β,IL-18 and adhesive molecule monocyte chemotactic protein-1(MCP-1)were detected by enzyme-linked immunosorbent assay(ELISA)after high Ca2+ stimulation.Results Ca2+ showed cytotoxicity for HK-2 cell growth and can promote apoptosis.The higher the Ca2+ concentration,the more toxicity and apoptosis rate for HK-2 cell growth.High concentration of Ca2+ can promote pyroptosis-like morphological changes in HK-2 cells,including loss of cell membrane integrity,release of contents and numerous intracellular vacuoles.Compared with the control group,the expression levels of ROS were sequentially increased in the 1.0 g/L CaCl2 group and the 2.0 g/L CaCl2 group,and the expression levels of pyroptosis-related genes NLRP3,Caspase-1,GSDMD,and the pyroptosis-associated inflammatory factors IL-1β and IL-18,as well as the adhesion molecules OPN,CD44 and MCP-1 were significantly increased(P＜0.05).Conclusion High Ca2+ treatment can cause oxidative stress damage in HK-2 cells to produce ROS,which activates NLRP3 inflammasome,leads to the activation of the classical pathway of pyroptosis and increase the adhesion of cells,and ultimately leads to the formation of kidney stones.

Sinomenine effectively inhibits interleukin-1beta-induced apoptosis in nucleus pulposus cells / 中国组织工程研究

BACKGROUND:Intervertebral disc degeneration is the basis of spinal degenerative diseases;however,there is no effective treatment. OBJECTIVE:To investigate whether sinomenine can inhibit interleukin-1β-induced apoptosis in nucleus pulposus cells and its molecular mechanism. METHODS:Rat nucleus pulposus cells were cultured in vitro by trypsin combined with type II collagenase digestion,and the cell growth curve was plotted.An appropriate sinomenine concentration was determined using the cell counting kit-8 kit.Nucleus pulposus cells were divided into control group,sinomenine group,interleukin-1β group,sinomenine+interleukin-1β group,zinc protoporphyrin group,zinc protoporphyrin+sinomenine group,zinc protoporphyrin+interleukin-1β group,and sinomenine+zinc protoporphyrin+interleukin-1β group.Proliferative activity,reactive oxygen species content,apoptosis rate,and heme oxygenase-1 expression in nucleus pulposus cells were detected. RESULTS AND CONCLUSION:The rat nucleus pulposus cells cultured in vitro were polygonal,triangular,and short wedge-shaped,and the cell growth showed an"S"curve.The cells grew slowly in the first 3 days of culture,rapidly in 4-6 days,and slowly again in 7-8 days.The cells then entered the"platform stage"where the number of cells no longer increased.The proliferative activity of myeloid cells showed no significant changes when the concentration of sinomenine was≤80 μmol/L(P>0.05).Interleukin-1β significantly reduced the proliferative activity of nucleus pulposus cells,increased the content of reactive oxygen species and led to apoptosis(P<0.01).Sinomenine intervention not only promoted heme oxygenase-1 expression(P<0.05)but also inhibited interleukin-1β-induced decrease in proliferative activity and increase in reactive oxygen species content and apoptosis rate in nucleus pulposus cells(P<0.05).These effects could be reversed by zinc protoporphyrin(P<0.01).

Prussian blue nanoparticles promote wound healing of diabetic skin / 中国组织工程研究

BACKGROUND:Inflammation,oxidative stress and bacterial infection are the main causes of delayed wound healing in diabetes.In recent years,various inorganic nanomaterials have been widely used in the treatment of skin wound healing due to their antibacterial activities,but their effects on anti-oxidation and anti-inflammation are limited. OBJECTIVE:To investigate the effect of Prussian blue nanoparticles on the wound repair of diabetes in terms of antioxidant,anti-inflammatory and photothermal antibacterial activities. METHODS:Prussian blue nanoparticles were prepared and characterized.(1)In vitro:The biocompatibility of Prussian blue nanoparticles with different concentrations was detected by MTT assay.The cytoprotective effect of Prussian blue nanoparticles and the intracellular reactive oxidative species level were examined under the condition of hydrogen peroxide.The ability of Prussian blue nanoparticles to decompose hydrogen peroxide and superoxide anion radicals was tested;the effect of Prussian blue nanoparticles on lipopolysaccharide-induced macrophage inflammation was investigated.The photothermal antibacterial activity of Prussian blue nanoparticles was detected by the plate colony counting method.(2)In vivo:ICR mice were intraperitoneally injected with streptozotocin to establish a diabetes mouse model.After the model was successfully established,a 6 mm wound was created on the back with a hole punch.There were the control group(no treatment),the Prussian blue group and the Prussian blue with light group.The wound healing and histomorphological changes were observed. RESULTS AND CONCLUSION:(1)In vitro:Prussian blue nanoparticles in 25-200 μg/mL were non-toxic to cells.Prussian blue nanoparticles had the extremely strong antioxidant capacity and mitigated the intracellular reactive oxidative species at a high oxidative stress environment,resulting in a pronounced cytoprotective effect.The Prussian blue nanoparticles not only exhibited hydrogen peroxide degradation activity but also showed strong superoxide scavenging ability.Prussian blue nanoparticles also displayed significant anti-inflammatory activity and extremely strong antibacterial ability after light irradiation.(2)In vivo:After 14 days,the wound sizes of the Prussian blue group and Prussian blue with light group were significantly reduced,and the healing speed of Prussian blue with light group was the fastest.Hematoxylin-eosin and Masson staining showed a lot of granulation tissue formation and collagen deposition in the Prussian blue group and the Prussian blue with light group,of which the Prussian blue with light group was the most.Immunofluorescence staining displayed that,compared with the control group,the expressions of α-SMA and CD31 were increased significantly in Prussian blue group and Prussian blue with light group(P<0.05),but F4/80 expression was decreased significantly in Prussian blue group and Prussian blue with light group(P<0.05),indicating more obvious improvement in the Prussian blue with light group.(3)These results showed that Prussian blue nanoparticles could promote the skin wound healing of the diabetes mouse model by exerting anti-inflammatory,antioxidant and antibacterial effects.

An insight into the mechanism of iron overload in knee osteoarthritis under the theory of blood stasis / 中国组织工程研究

BACKGROUND:Knee osteoarthritis is a common disease in middle-aged and elderly people.It is a kind of disease that seriously affects the quality of life of patients and even has the risk of disability.Therefore,the pathogenesis and treatment of knee osteoarthritis have become the focus of research.In Chinese medicine,knee osteoarthritis is often treated as"biness,"which is closely related to"biness"caused by blood stasis and blood vessels blocking collaterals in the theory of"blood stasis"in traditional Chinese medicine.Iron overload is a kind of pathological state caused by iron metabolism disorder,which highly coincides with the pathogenic characteristics and clinical manifestations of the"blood stasis"theory of traditional Chinese medicine,and is a risk factor that promotes the development of knee osteoarthritis. OBJECTIVE:Based on the"blood stasis"theory,to summarize the effects of iron overload on cartilage metabolism and subchondral bone reconstruction,to lay a new theoretical foundation for the treatment of knee osteoarthritis with traditional Chinese medicine,and to explore the therapeutic effect of traditional Chinese medicine for promoting blood circulation after interfering with bone tissue. METHODS:CNKI,WanFang database,PubMed and Web of Science databases were searched for relevant literature.The Chinese search terms were"ferroptosis,iron,iron overload,osteoarthritis,blood stasis"and the English search terms were"ferroptosis,iron,iron overload,osteoarthritis,TCM."In the end,76 articles were included for further review. RESULTS AND CONCLUSION:First of all,we explored the potential of the"blood stasis"theory in treating knee osteoarthritis,and found that"blood stasis"is a crucial part in the progress of knee osteoarthritis,indicating that the"blood stasis"theory is the key to the treatment of knee osteoarthritis in traditional Chinese medicine.Secondly,"blood stasis"and iron overload have a high degree of similarity in pathogenic factors,clinical manifestations,and pathogenic characteristics,suggesting the possibility of"blood stasis"theory in treating iron overload.This finding reminds us that iron overload may be an important mechanistic basis for the"blood stasis"theory in the treatment of knee osteoarthritis.The extracts of blood-activating drugs can relieve iron overload and treat knee osteoarthritis,but the specific mechanism is still unclear.Therefore,we believe that the relationship between"blood stasis"theory and iron overload and related mechanisms are important research directions for knee osteoarthritis in the future.The related mechanism of"blood stasis"theory to alleviate iron overload and then treat knee osteoarthritis also provides a theoretical basis for the modernization of traditional Chinese medicine,such as the development of new drugs and innovative usage,and has certain guiding significance for clinical practice.

Role and mechanism of reactive oxygen species in tendinopathy / 中国组织工程研究

BACKGROUND:Reactive oxygen species may be closely related to the occurrence and development of tendinopathy,but its exact role and related signal transduction mechanism have not been fully summarized. OBJECTIVE:To review current clinical or preclinical original studies,summarize the role of reactive oxygen species in tendinopathy and related signal transduction pathways and to explore its characteristics and whether there is a unified downstream pathway. METHODS:Relevant original studies in PubMed,Embase,Web of Science,as well as CNKI,WanFang,and VIP databases were searched by computer and the search results were screened and excluded according to the inclusion criteria.Ninety articles were finally included for review and analysis. RESULTS AND CONCLUSION:Reactive oxygen species affects the direction of tendon healing by simultaneously acting on tendon cells and the extracellular matrix,and it exhibits a bifacial effect in the treatment of tendinopathy.Concentration of reactive oxygen species may be the key to determining its direction of action.The possibility that low-dose reactive oxygen species can participate in the normal physiological healing of tendons or that tendon tissues are adaptive to stimulations may be the underlying mechanism that produces this characteristic effect.Reactive oxygen species affect the composition and structure of the extracellular matrix and normal tendon repair as well as maintain viability in response to external stimulations through matrix metalloproteinases,mitogen-activated protein kinases,mitochondrial apoptosis,the forkhead transcription factor O family,autophagy,inflammation,and antioxidant signaling pathways.Different reactive oxygen species stimulation intensities,durations,and external environments may cause different alterations in downstream molecular pathways and thus have different effects on the tendon.Due to the large gap in the number of literature included in the evaluation of the positive and negative effects of reactive oxygen species,it may cause some analytical error in the search for factors behind the characteristics of the action of reactive oxygen species in tendon.In addition,most experimental intervention conditions and results of interest are relatively homogeneous;therefore,the temporal and quantitative mechanisms of reactive oxygen species and the synergistic effects with other intervention factors have not been clarified,and the overall system of molecular actions of reactive oxygen species in tendinopathy has not been constructed.To conclude,reactive oxygen species might be involved in the treatment and prevention of tendinopathies as a beneficial factor in the future,and facilitate the exploration of oxidative stress signaling pathways and overall molecular action systems in tendinopathies thereafter,as well as lay the foundation for research on the therapeutic strategies of different antioxidants in tendinopathies to better prevent and treat tendon injury and degeneration.

Ginsenoside Rg1 inhibits H2O2-induced apoptosis and autophagy in human periodontal ligament cells / 中国组织工程研究

BACKGROUND:Our previous studies have confirmed that H2O2 and ginsenoside Rg1 can cause changes in reactive oxygen species levels in human periodontal ligament cells,but the correlation of reactive oxygen species with apoptosis and autophagy remains unclear. OBJECTIVE:To explore the effect and mechanism of ginsenoside Rg1 on H2O2-induced apoptosis and autophagy of human periodontal ligament cells. METHODS:Human periodontal ligament cells were divided into control group,H2O2 group and H2O2+Rg1 group.Ginsenoside Rg1(50 μmol/L)was pre-incubated for 24 hours and H2O2 was treated for 2 hours(500 μmol/L).CCK-8 assay was used to detect the proliferation ability of the cells.A fluorescent probe DCFH-DA was used to detect the reactive oxygen species level of the cells.qRT-PCR and western blot assay were used to detect heme oxygenase 1,apoptosis-related factor Caspase-3,Bax,anti-apoptotic factor Bcl-2,autophagy Beclin-1,P62,LC3,pathway-related factors phosphatidylinositol-3-kinase(PI3K),protein kinase B(AKT),mammalian target of rapamycin(mTOR)mRNA and protein expression. RESULTS AND CONCLUSION:(1)Compared with the control group,the proliferation activity of human periodontal ligament cells in the H2O2 group decreased.Compared with the H2O2 group,the proliferation activity of human periodontal ligament cells increased in the H2O2+Rg1 group.(2)Compared with the control group,the expression of reactive oxygen species and heme oxygenase-1 increased in the H2O2 group.Compared with the H2O2 group,the expression of reactive oxygen species and heme oxygenase-1 decreased in the H2O2+Rg1 group.(3)Compared with the control group,the expression of Caspase-3,Bax,Beclin-1 and LC3 increased,while the expression of Bcl-2 and P62 decreased in human periodontal ligament cells of the H2O2 group.Compared with the H2O2 group,the expressions of Caspase-3,Bax,Beclin-1 and LC3 decreased,and the expressions of Bcl-2 and P62 increased in the H2O2+Rg1 group.(4)Compared with the control group,the expressions of PI3K,AKT and mTOR decreased in human periodontal ligament cells of the H2O2 group.Compared with the H2O2 group,the expressions of PI3K,AKT and mTOR increased in human periodontal ligament cells of the H2O2+Rg1 group.(5)These results suggest that ginsenoside Rg1 can inhibit H2O2-induced apoptosis and autophagy in human periodontal ligament cells by reducing the content of reactive oxygen species and down-regulating the related factor expression of the PI3K/AKT/mTOR pathway.

The role of reactive oxygen species in periodontitis and periodontal tissue regeneration / 中国组织工程研究

BACKGROUND:Reactive oxygen species is a double-edged sword in the development of periodontitis and the regeneration of periodontal tissue.Low concentration of reactive oxygen species induces the differentiation of periodontal fibroblasts,and excessive reactive oxygen species will cause damage to periodontal tissue.In the process of inflammation,the accumulation of reactive oxygen species in periodontal tissue induces damage to cells and tissues through a variety of signaling pathways or through redox reactions. OBJECTIVE:To review the double-edged sword effect of reactive oxygen species in periodontitis and periodontal tissue regeneration,thereby providing potential targets and treatment ideas for the clinical treatment of periodontitis and periodontal tissue regeneration. METHODS:Databases of CNKI and PubMed were searched for relevant articles published from April 1990 to April 2023 with the key words of"periodontal tissue engineering,periodontal defect,regeneration of periodontal tissue,chronic periodontitis,reactive oxygen species,oxidative stress,antioxidative stress,oxidative injuries,free radicals,reactive nitrogen species"in Chinese and English,respectively.By reading the titles and abstracts,repetitive studies or irrelevant literatures were excluded.Finally,77 articles were included for review. RESULTS AND CONCLUSION:Reactive oxygen species are a kind of free radicals with high reactivity.When bacteria invade,reactive oxygen species are released in large quantities by the respiratory explosion of neutrophils and play a double-edged sword role in the body through their redox reactions or as pleiotropic physiological signal transmitters.In periodontitis,low concentrations of reactive oxygen species can kill invading pathogenic bacteria,but high concentrations of reactive oxygen species promote the secretion of inflammatory factors through JNK,RANK,Wnt/β-Catenin and other pathways,promote immune damage or directly damage tissues through oxidative reactions or through other ways to aggravate periodontitis.In the process of periodontal tissue regeneration,low concentrations of reactive oxygen species can promote the proliferation and differentiation of periodontal ligament stem cells through Nrf2 and other pathways and can promote the secretion of vascular endothelial growth factor to promote vascular regeneration.This provides seeds and a nutrient environment for periodontal tissue regeneration,which is extremely important for promoting periodontal tissue regeneration.However,high concentrations of reactive oxygen species will reduce the activity of periodontal ligament stem cells and damage endothelial cells,which are not conducive to vascular regeneration.This will inhibit wound healing and periodontal tissue regeneration.Therefore,it is important to explore the role of reactive oxygen species in the development of periodontitis and periodontal tissue regeneration and to discover the potential mechanism of its action and to explore the appropriate concentration for its role in reducing periodontal inflammation and promoting periodontal tissue regeneration for the future treatment of periodontitis and periodontal tissue regeneration in clinical practice.Using reactive oxygen species as a target to explore ways to reduce periodontal inflammation and promote periodontal ligament stem cell activity and vascular regeneration may become a clinically effective method for treating periodontitis and promoting periodontal tissue regeneration.

Ferroptosis in bone diseases:therapeutic targets of osteoporosis / 中国组织工程研究

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.

Berberine promotes osteogenic differentiation of bone marrow mesenchymal stem cells in a high-glucose environment / 中国组织工程研究

BACKGROUND:The implant osseointegration rate of patients with diabetes is low,and the failure rate is high,which seriously affects the quality of life.It is urgent to improve the implant osseointegration of patients with diabetes by effective means to elevate the success rate.Exploring the effect of berberine on the osteogenic differentiation of bone marrow mesenchymal stem cells under a high-glucose environment and its specific mechanism will provide effective theoretical support for solving the above problems. OBJECTIVE:To explore the effect of natural extract berberine on the osteogenic differentiation of rat bone marrow mesenchymal stem cells under the high-glucose microenvironment. METHODS:Bone marrow mesenchymal stem cells of SD rats were cultured by the whole bone marrow adherence method.CCK-8 assay was used to detect the effects of different concentrations of berberine on the proliferation of bone marrow mesenchymal stem cells under the high-glucose environment and to screen out the optimal berberine concentration.The expressions of Runx2 and Osx were detected by alkaline phosphatase activity,alicarin red staining and PCR to determine the effect of berberine on osteogenic differentiation of bone marrow mesymal stem cells under the high-glucose environment.To further explore the underlying mechanism,we introduced the AMPK-specific inhibitor Dorsomorphin and used a DCFH-DA reactive oxygen species fluorescent probe to examine reactive oxygen species levels.The p-AMPK expression was also determined by western blot assay. RESULTS AND CONCLUSION:(1)10 μmol/L was the optimal concentration of berberine to promote bone marrow mesenchymal stem cell proliferation.(2)Alberberine promoted alkaline phosphatase viability of bone marrow mesenchymal stem cells and mineralized nodule formation in a high-glucose microenvironment.(3)Alberberine promoted the expression of Runx2 and OSx in a high-glucose microenvironment.(4)Alberensine effectively inhibited the reactive oxygen species level of bone marrow mesenchymal stem cells in a high-glucose environment.(5)The effects of berberine on promoting bone marrow mesenchymal stem cell osteogenesis and inhibition of reactive oxygen species were reversed by the AMPK inhibitor.(6)Berberine activated AMPK and promoted p-AMPK expression.(7)The above results indicate that berberine(10 μmol/L)promotes the osteogenic differentiation of bone marrow mesenchymal stem cells in a high-glucose environment by activating AMPK and reducing intracellular reactive oxygen species levels.

Vitamin D3 attenuates high-glucose exposure-induced oxidative stress to promote osteogenic differentiation of human umbilical cord mesenchymal stem cells / 中国组织工程研究

BACKGROUND:Diabetic osteoporosis is gaining public attention.However,few studies have reported the effect of a high-glucose environment on the osteogenic differentiation of human umbilical cord mesenchymal stem cells and the corresponding therapeutic strategies. OBJECTIVE:To investigate whether vitamin D3 can restore the osteogenic differentiation potential of human umbilical cord mesenchymal stem cells in a high-glucose environment. METHODS:The viability of human umbilical cord mesenchymal stem cells was detected by CCK-8 assay to screen the appropriate vitamin D3 intervention concentration.Under the high-glucose environment,RT-qPCR,western blot assay,immunofluorescence,JC-1 mitochondrial membrane potential,alizarin red staining,and β-galactosidase staining were used to evaluate the osteogenic differentiation potential,intracellular reactive oxygen species accumulation,mitochondrial membrane potential alteration,and cell senescence of human umbilical cord mesenchymal stem cells after vitamin D3 intervention.The underlying mechanism was also discussed. RESULTS AND CONCLUSION:(1)Vitamin D3 significantly promoted the proliferation of human umbilical cord mesenchymal stem cells in the range of 0.1 μmol/L to 1 mmol/L.(2)High-glucose environment down-regulated the mRNA and protein level expressions of osteogenic-related genes α1-I collagen,alkaline phosphatase,Runt-associated transcription factor 2,and osteocalcin in human umbilical cord mesenchymal stem cells,which induced oxidative stress and cellular senescence.(3)Vitamin D3 at an intervention concentration of 10 μmol/L significantly restored the osteogenic phenotype of human umbilical cord mesenchymal stem cells under high-glucose conditions and attenuated intracellular oxidative stress and cellular senescence by activating the Nrf2/HO-1 signaling pathway.(4)These findings suggested that the osteogenic differentiation ability of human umbilical cord mesenchymal stem cells was reduced in the high-glucose environment,and vitamin D3 could partially improve their osteogenic differentiation ability and reduce cell damage.