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1.
Bol. latinoam. Caribe plantas med. aromát ; 23(4): 516-522, jul. 2024. graf, ilus
Article in English | LILACS | ID: biblio-1538029

ABSTRACT

This article aimed to discuss the protection of trans - nerolidol on vascular endothelial cells (ECs) injured by lipopolysac charides. ECs were divided into four groups: normal, model, low and high dose trans - nerolidol treatment groups. The cell survival rate and the contents of NO in the cell culture supernatant were determined. The protein expression and transcript level of pe roxisome proliferator - activated receptor - γ (PPARγ), endothelial nitric oxide synthase (eNOS), and inducible nitric oxide synthase (iNOS) were determined by western blotting and RT - PCR respectively. Compared with the normal group, cell livability, protein e xpression and mRNA transcript level of PPARγ and eNOS decreased, NO contents, protein expression and mRNA transcript tlevel of iNOS increased in model group significantly. Compared with model group, all the changes recovered in different degree in treatmen t groups. Hence, it was concluded that trans - nerolidol can alleviate the ECs injuryby the regulation of iNOS/eNOS through activating PPARγ in a dose - dependent manner


Este artículo tiene como objetivo discutir la protección del trans - nerolidol en las células endoteliales vasculares (CE) dañadas por lipopolisacáridos. Las CE se di vidieron en cuatro grupos: normal, modelo, grupos de tratamiento con trans - nerolidol de baja y alta dosis. Se determinó la tasa de supervivencia de las células y los contenidos de óxido nítrico (NO) en el sobrenadante del cultivo celular. La expresión de p roteínas y el nivel de transcripción del receptor activado por proliferadores de peroxisomas - γ (PPARγ), el óxido nítrico sint et asa endotelial (eNOS) y el óxido nítrico sint et asa inducible (iNOS) se determinaron mediante western blot y RT - PCR, respectivamen te. En comparación con el grupo normal, la viabilidad celular, la expresión de proteínas y el nivel de transcripción de PPARγ y eNOS disminuyeron, los contenidos de NO, la expresión de proteínas y el nivel de transcripción de iNOS aumentaron significativam ente en el grupo modelo. En comparación con el grupo modelo, todos los cambios se recuperaron en diferentes grados en los grupos de tratamiento. Por lo tanto, se concluyó que el trans - nerolidol puede aliviar el daño en las CE regulando iNOS/eNOS a través d e la activación de PPARγ de manera dependiente de la dosis.


Subject(s)
Sesquiterpenes/administration & dosage , Vascular Diseases/drug therapy , Endothelium, Vascular/drug effects , Endothelium, Vascular/injuries , Cell Survival , Lipopolysaccharides/toxicity , Blotting, Western , Nitric Oxide Synthase , Real-Time Polymerase Chain Reaction
2.
Recent Advances in Ophthalmology ; (6): 30-34,38, 2024.
Article in Chinese | WPRIM | ID: wpr-1022709

ABSTRACT

Objective To investigate the anti-inflammatory action of Celastrol in the ocular tissues of mice with ex-perimental autoimmune uveitis(EAU)and its effect on microglia polarization.Methods A total of 36 healthy B10.RⅢmice at 6-8 weeks of age were selected and randomly divided into the normal control group,EAU solvent control group and Celastrol intervention group,with 12 mice in each group.The interphotoreceptor retinoid-binding protein(IRBP)161-180 and Freund's complete adjuvant were mixed by thorough emulsification and injected subcutaneously into the bilateral thighs and tails of mice in the EAU solvent control group and the Celastrol intervention group with a total volume of 200 μL and 50 μg IRBP 161-180 in each mouse.On 7-14 days after immunization,mice in the Celastrol intervention group received a daily intraperitoneal injection of 0.5 mg·kg-1 Celastrol,and mice in the EAU solvent control group were injected with an equivalent dose of sterile Phosphate Buffered Saline solution.On the 14th day after immunization,the anterior segment of mice in each group was observed by slit-lamp microscope and Hematoxylin and Eosin(HE)staining of tissue sections was performed;the clinical and histopathological scores of mice in each group were obtained by reference to the Caspi grading standards;immunofluorescence staining was used to observe the activation of microglia in the eyes of mice;Western blot was used to detect the protein expression levels of inducible nitric oxide synthase(iNOS)and arginase-1(Argl)in the reti-na;quantitative real-time PCR was used to detect the relative mRNA expression of inflammatory factors in the retina,such as tumor necrosis factor(TNF)-α,interleukin(IL)-1β and IL-6.GraphPad Prism 9.0 was used for data analysis.Results On the 14th day after immunization,it was observed by the slit-lamp microscope that the anterior segment of mice in the EAU solvent control group was markedly congested with dilated iris blood vessels,corneal edema,and anterior chamber exudation;the inflammation in the anterior segment of mice in the Celastrol intervention group was markedly at-tenuated,and the iris blood vessels were seen to be mildly congested.Compared with the normal control group,the clini-cal scores of mice in the EAU solvent control group and the Celastrol intervention group were significantly elevated(both P<0.05);the clinical scores of mice in the Celastrol intervention group were lower than those in the EAU solvent control group(P<0.05).HE staining results showed that on the 14th day after immunization,mice in the EAU solvent control group showed severe retinal folds and detachment with diffuse infiltration of inflammatory cells,while mice in the Celastrol intervention group showed slight structural damage to the retina and a small amount of inflammatory cell infiltration.Com-pared with the normal control group,the histopathological scores of mice in the EAU solvent control group and the Celas-trol intervention group were significantly elevated(both P<0.05);the histopathological scores of mice in the Celastrol in-tervention group were lower than those in the EAU solvent control group(P<0.05).The intraocular Iba1+cell densities of mice in the normal control,EAU solvent control and Celastrol intervention groups were(1.00±0.12)%,(36.07± 4.57)%,and(1.83±0.36)%,respectively.Compared with the normal control group,the number of Iba1+cells in the eyes of mice in the EAU solvent control group and the Celastrol intervention group significantly increased(both P<0.05);compared with the EAU solvent control group,the number of Iba1+cells in the eyes of mice in the Celastrol intervention group was significantly reduced(P<0.05).Compared with the normal control group,the expression levels of iNOS and Arg1 proteins in the retinas of mice in the EAU solvent control group were significantly elevated(both P<0.01);compared with the EAU solvent control group,the expression of iNOS protein in the retinas of mice in the Celastrol intervention group was significantly reduced(P<0.01).Compared with the normal control group,the relative mRNA expressions of TNF-α.IL-1β,and IL-6 in the retinas of mice in the EAU solvent control group was significantly elevated(all P<0.05);compared with the EAU solvent control group,the relative mRNA expressions of TNF-α,IL-1 β,and IL-6 in the retinas of mice in the Celastrol intervention group significantly decreased(all P<0.05).Conclusion Celastrol inhibits Ml microglia activation and reduces the production of retinal inflammatory factors TNF-α,IL-1 β and IL-6 in EAU mice,thereby attenuating the in-flammatory reaction.

3.
Article in Chinese | WPRIM | ID: wpr-996153

ABSTRACT

Objective:To observe the effects of acupuncture at Houxi(SI3)and Huantiao(GB30)on the expression levels of nuclear factor kappa B(NF-κB),inducible nitric oxide synthase(iNOS),and nitric oxide(NO)of NF-κB inflammatory signaling pathway in L5 spinal nerve root of lumbar disc herniation(LDH)model rats and explore the mechanism of acupuncture in LDH treatment.Methods:Forty specific-pathogen-free healthy male Sprague-Dawley rats were randomly divided into a sham operation group,a model group,acupuncture group 1,and acupuncture group 2,with 10 rats in each group.The non-compression nucleus protrusion model was made by puncturing L4-L5 spinous process space and injecting autologous nucleus suspension.Acupuncture at bilateral Shenshu(BL23),Dachangshu(BL25),and Weizhong(BL40)was carried out in acupuncture group 1,and acupuncture at bilateral Houxi(SI3)and Huantiao(GB30)in acupuncture group 2.All rats were treated with balanced reinforcing and reducing needling manipulations,and the needles were retained for 30 min/time with one episode of needling manipulation every 10 min,once a day,14 times in total.The threshold value of paw withdrawal pain was measured by a thermal stimulation pain instrument;the serum NF-κB,iNOS,and NO levels were measured by enzyme-linked immunosorbent assay.The pathomorphological changes of spinal nerve roots were observed by hematoxylin-eosin(HE)staining;quantitative reverse transcription polymerase chain reaction was used to detect iNOS mRNA expression in spinal nerve roots;the NF-κB and iNOS protein expression in spinal nerve roots was detected by the immunofluorescence method.Results:Compared with the sham operation group,the threshold of paw withdrawal pain in the model group was decreased,and the expression levels of serum NF-κB,iNOS,and NO were increased;HE staining showed many degenerated and dissolved Schwann cells in spinal nerve roots with vacuolar changes;meanwhile,the expression levels of NF-κB and iNOS proteins,and the iNOS mRNA in spinal nerve roots were increased.Compared with the model group,the paw withdrawal pain thresholds in acupuncture group 1 and acupuncture group 2 were increased,and the increase in acupuncture group 2 was greater(P<0.05);the expression levels of serum NF-κB,iNOS,and NO in acupuncture group 1 and acupuncture group 2 were decreased,especially in acupuncture group 2(P<0.01);the vacuolar changes of spinal nerve roots,and the degeneration and lysis of Schwann cells in acupuncture group 1 and acupuncture group 2 were decreased,which were more obvious in acupuncture group 2;the NF-κB and iNOS protein expression and the iNOS mRNA expression levels in spinal nerve roots of acupuncture group 1 and acupuncture group 2 were decreased,especially in acupuncture group 2(P<0.01).Conclusion:Acupuncture at Houxi(SI3)and Huantiao(GB30)can improve the morphology of spinal nerve roots,inhibit the NF-κB and iNOS protein expression levels in spinal nerve roots and the serum NO level,and relieve the pain caused by inflammation of spinal nerve roots,which may be one of the mechanisms of acupuncture in LDH treatment.

4.
Article in Chinese | WPRIM | ID: wpr-996811

ABSTRACT

ObjectiveTo decipher the mechanism of modified Sanpiantang in the treatment of nitroglycerin-induced migraine in rats. MethodSeventy-two Wistar rats were randomized into the control, model (nitroglycerin, 10 mg·kg-1), positive control (rizatriptan, 0.89 mg·kg-1), and high- (12.96 g·kg-1), medium- (6.48 g·kg-1), and low-dose (3.24 g·kg-1) modified Sanpiantang groups. The rat model of migraine was established by intraperitoneal injection of 10 mg·kg-1 nitroglycerin. The behavioral test was carried out to measure the mechanical pain thresholds (MPT) of the periorbital region and hindpaw after successful modeling. The serum levels of nitric oxide (NO), tumor necrosis factor-α (TNF-α), interferon-γ (IFN-γ), and interleukin-1β (IL-1β) in rats were determined by enzyme-linked immunosorbent assay (ELISA). Immunohistochemistry (IHC) was employed to determine the expression of inducible nitric oxide synthase (iNOS) in the trigeminal nucleus caudalis (TNC). Western blot was employed to determine the protein levels of iNOS and phospho-p38 mitogen-activated protein kinase (p-p38 MAPK) in the TNC. Real-time fluorescence quantitative polymerase chain reaction(Real-time PCR) was performed to measure the mRNA levels of iNOS, p38 MAPK, and IL-1β in the TNC. ResultCompared with the control group, the model group showed decreased MPT (P<0.01), elevated serum levels of NO, TNF-α, IFN-γ, and IL-1β (P<0.01), and up-regulated expression levels of p38 MAPK, iNOS, and IL-1β in the TNC (P<0.05, P<0.01). Compared with the model group, modified Sanpiantang increased the MPT (P<0.05), and the medium-dose group showed the most significant effect (P<0.01). In addition, modified Sanpiantang down-regulated the mRNA levels of iNOS, p38 MAPK, and IL-1β and the protein levels of p-p38 MAPK and iNOS in the TNC of migraine rats (P<0.05, P<0.01) and lowered the serum levels of NO, TNF-α, IFN-γ, and IL-1β (P<0.05, P<0.01). ConclusionModified Sanpiantang may treat migraine by down-regulating the expression of pro-inflammatory factors such as NO, TNF-α, IFN-γ, and IL-1β in the p38 MAPK/iNOS signaling pathway to reduce the neurogenic inflammation.

5.
Article in Chinese | WPRIM | ID: wpr-978450

ABSTRACT

ObjectiveTo observe the glucose-lowering, insulin resistance-improving, and anti-inflammatory effects of flavonoids from mulberry leaves (FML) and explore their underlying mechanism. MethodMale db/db mice aged 6-7 weeks were randomly divided into a model group, a high-dose FML group (1.00 g·kg·d-1), and a low-dose FML group (0.50 g·kg-1·d-1). C57BL mice of the same age were assigned to the normal group. After six weeks of intervention, fasting blood glucose (FBG), serum fasting insulin levels (Fins), interleukin-6 (IL-6), tumor necrosis factor-α (TNF-α), free fatty acid (FFA), blood creatinine (SCr), blood urea nitrogen (BUN), and aspartate aminotransferase (AST) levels were measured, and the homeostasis model assessment of insulin resistance (HOMA-IR) was calculated. Superoxide dismutase (SOD), glutathione peroxidase (GSH-Px), and catalase activities in the liver were measured. Morphological changes in the liver were assessed by hematoxylin-eosin (HE) staining. The protein expression of cyclooxygenase-2 (COX-2), inducible nitric oxide synthase (iNOS), and nuclear factor-κB (NF-κB) in the liver was detected by Western blot. ResultCompared with the model group, the high-dose and low-dose FML groups showed significant reductions in FBG, Fins, HOMA-IR, IL-6, TNF-α, and FFA levels (P<0.05, P<0.01), and increased levels of SOD, GSH-Px, and catalase in the liver (P<0.05, P<0.01). HE staining of the liver in the FML groups showed improved arrangement of hepatocytes, reduced inflammatory cell infiltration, and alleviated cellular steatosis compared with the model group. The protein expression of COX-2, iNOS, and NF-κB in the liver significantly decreased in the FML groups as compared with that in the model group (P<0.05, P<0.01). ConclusionFML have glucose-lowering and insulin resistance-improving effect, which may be attributed to their regulation of the NF-κB pathway in the liver of diabetic mice, leading to the suppression of the release of COX-2, iNOS, and inflammatory cytokines, thereby improving the inflammatory state.

6.
Article in Chinese | WPRIM | ID: wpr-991110

ABSTRACT

Glycyrrhizae Radix et Rhizoma,a traditional Chinese medicine also known as Gan Cao(GC),is frequently included in clinical prescriptions for the treatment of pneumonia.However,the pharmacological com-ponents of GC for pneumonia treatment are rarely explored.Gan An He Ji oral liquid(GAHJ)has a simple composition and contains GC liquid extracts and paregoric,and has been used clinically for many years.Therefore,GAHJ was selected as a compound preparation for the study of GC in the treatment of pneumonia.We conducted an in vivo study of patients with pneumonia undergoing GAHJ treatments for three days.Using the intelligent mass spectrometry data-processing technologies to analyze the meta-bolism of GC in vivo,we obtained 168 related components of GC in humans,consisting of 24 prototype components and 144 metabolites,with 135 compounds screened in plasma and 82 in urine.After analysis of the metabolic transformation relationship and relative exposure,six components(liquiritin,liquiritigenin,glycyrrhizin,glycyrrhetinic acid,daidzin,and formononetin)were selected as potential effective components.The experimental results based on two animal pneumonia models and the in-flammatory cell model showed that the mixture of these six components was effective in the treatment of pneumonia and lung injury and could effectively downregulate the level of inducible nitric oxide synthase(iNOS).Interestingly,glycyrrhetinic acid exhibited the strongest inhibition on iNOS and the highest exposure in vivo.The following molecular dynamic simulations indicated a strong bond between glycyrrhetinic acid and iNOS.Thus,the current study provides a pharmaceutical basis for GC and reveals the possible corresponding mechanisms in pneumonia treatment.

7.
China Tropical Medicine ; (12): 1179-2022.
Article in Chinese | WPRIM | ID: wpr-972143

ABSTRACT

@#Abstract: Objective To explore the correlation between the levels of silent information regulator 1 (SIRT1) and forkhead box protein O3 (FOXO3) in peripheral blood mononuclear cells of patients with active pulmonary tuberculosis (APTB) and macrophage-related cytokines-inducible nitric oxide synthase (iNOS) and arginase-1 (Arg-1). Methods A total of 64 APTB patients who were treated in Yubei Hospital, the First Affiliated Hospital of Chongqing Medical University from January 2020 to December 2021 were gathered as the APTB group, 59 people with latent tuberculosis infection (LTBI) were gathered as the LTBI group, and 62 healthy people were gathered as the control group. Quantitative real-time PCR (qPCR) method was performed to measure the levels of SIRT1 mRNA and FOXO3 mRNA in peripheral blood mononuclear cells. The enzyme-linked immunosorbent assay (ELISA) was performed to measure serum iNOS and Arg-1 levels; ROC curve was used to analyze the value of SIRT1 mRNA and FOXO3 mRNA levels in the differential diagnosis of LTBI and APTB; Pearson correlation was performed to analyze the correlation of SIRT1 mRNA and FOXO3 mRNA in peripheral blood mononuclear cells of APTB patients with serum iNOS and Arg-1 levels. Results The levels of SIRT1 mRNA, FOXO3 mRNA and serum iNOS in peripheral blood mononuclear cells decreased in control group, LTBI group and APTB group, and the level of serum Arg-1 increased in turn (P<0.05). The AUCs of SIRT1 mRNA and FOXO3 mRNA in differential diagnosis of LTBI and APTB were 0.876 and 0.887, respectively, the sensitivity was 71.2% and 76.3%, and the specificity was 96.9% and 90.6% respectively. The levels of SIRT1 mRNA and FOXO3 mRNA in peripheral blood mononuclear cells of APTB patients were positively correlated (r=0.500, P<0.05), and they were positively correlated with serum iNOS and negatively correlated with serum Arg-1 (P<0.05). The SIRT1 mRNA, FOXO3 mRNA and serum iNOS in peripheral blood mononuclear cells of APTB patients after 6 months of treatment were higher than those before treatment, and serum Arg-1 was lower than before treatment (P<0.05). Conclusions The levels of SIRT1 mRNA and FOXO3 mRNA in peripheral blood mononuclear cells of APTB patients are low, and they are positively correlated with macrophage-related cytokine iNOS and negatively correlated with Arg-1.

8.
Article in Chinese | WPRIM | ID: wpr-930228

ABSTRACT

Objective:To observe the relationship between inducible carbon monoxide synthase (iNOS) and delayed encephalopathy after acute carbon monoxide poisoning (DEACMP), and explore its mechanism of action in DEACMP.Methods:This study was designed as prospective cohort study. Patients with acute carbon monoxide poisoning who met the diagnostic criteria and were admitted to Emergency Intensive Care Unit(EICU) of our hospital from June 2019 to June 2021 were selected as subjects. Patients were divided into the DEACMP group and non-DEACMP group according to the occurrence of DEACMP. Serum samples were collected on the first 24 h after admission and on day 7 and 14 after admission, and the serum nitric oxide (NO), neuronal nitric oxide synthase (nNOS), inducible carbon monoxide synthase (iNOS), and endothelial nitric oxide synthase (eNOS) level were measured by enzyme-linked immunosorbent assay. The generalized estimating equation was used to estimate the difference of NO, nNOS, iNOS and eNOS between DEACMP and non-DEACMP patients.Results:A total of 78 patients with carbon monoxide poisoning were included in our study finally, including 49 (62.82%) males and 29 (37.18%) females, with an average age of (53.96±14.95) years, 20 (25.64%) patients with DEACMP, and 1 (1.28%) death. Univariate analysis showed that patients with DEACMP had an average increase of 3 h (95% CI: 1.00, 5.00) in carbon monoxide exposure time and a 5-point decrease in GCS score (95% CI: 1.00, 6.00) than the patients without DEACMP, and the proportion of patients with severe carbon monoxide poisoning in the DEACMP group was higher than that of the non-DEACMP group (90.00% vs. 32.76%). According to the analysis of generalized estimation equation, on day 7 and 14 after admission, Compared with non-DEACMP patients, neither by performing unadjusted nor adjusted analysis with the iNOS of DEACMP patients was significantly higher than that in non-DEACMP patients regardless of whether exposure time, GCS score, coma time or severity of carbon monoxide poisoning were adjusted or not ( P <0.01 or P <0.05). Except for the level of nNOS in the GEE model adjusted with carbon monoxide exposure time, the levels of NO, nNOS and eNOS showed no significant difference between DEACMP and non-DEACMP patients ( P >0.05). Conclusions:The expression of iNOS level is increased in DEACMP patients, and its continuous expression may be involved in the pathogenesis of DEACMP.

9.
Acta Anatomica Sinica ; (6): 340-346, 2022.
Article in Chinese | WPRIM | ID: wpr-1015322

ABSTRACT

Objective To explore the relationship between the content of neuropeptide Y (NPY), calcitonin gene-related peptide (CGRP), inducible nitric oxide synthase (iNOS) and the changes of the corresponding receptors in myocardial tissue and the development of heart failure. Methods Doxorubicin was used to prepare rat heart failure models(10 rats), control group (10 rats), Cardiac ultrasound testing evaluated its cardiac function, the distribution of nerve fibers was detected by nerve staining kit method, the tissue expression levels of NPY, CGRP, iNOS and neuropeptide Y receptor Y1 (NPY1R), receptor activity modifying protein 1 (RAMP1), guanylate cyclase beta 1(GCYB1) were observed by immunofluorescence and immunohistochemical staining method. Western blotting was used to detect the protein expression levels of NPY, CGRP, iNOS and NPY1R, RAMP1, GCYB1 receptors in myocardial tissue. Results Compared with the control group, the protein expression level and tissue expression distribution of CGRP in the myocardial tissue of the heart failure group were significantly decreased (P<0.05), while the density of nerve fibers, the protein expression level and tissue expression distribution of NPY and iNOS were significantly increased (P<0.05). The protein expression levels of GCYB1 and RAMP1 in myocardial tissue of the heart failure group were significantly increased, while the protein expression levels of NPY1R were significantly decreased (P<0.05). Conclusion Heart failure can lead to remodeling of the content and distribution of nerve fibers, CGRP, NPY, iNOS and their receptors in myocardial tissue, which may be one of the causes of cardiac innervation disorders.

10.
Acta Anatomica Sinica ; (6): 288-294, 2022.
Article in Chinese | WPRIM | ID: wpr-1015337

ABSTRACT

Objective To testify the link between the increase of inducible nitric oxide synthase (iNOS) and monocarboxylate transporter 1 (MCT1) expression in the M1 phenotype microglia under ischemic condition. Methods Photothrombosic ischemia stroke model was applied in 6 mice. BV2 cells were treated with low glucose medium contained 5mmol/L glucose for 2 hours. Immunofluorescent staining and Western blotting were used to check the responses from microglia in the mouse brains subjected to ischemia and BV2 cells were treated with low-glucose treatment. Application of RNA interference or plasmid transfection were used to regulate the MCT1 expression in BV2 cells. Results The expression levels of iNOS, MCT1 and arginase-1 (ARG1) increased in the ischemic side compared to the non-ischemic side of mice brain(P<0.01). In the BV2 cells exposed to low-glucose condition, iNOS and MCT1 levels increased(P<0.001), whereas ARG1 level decreased(P<0.01). RNA interference interfered the expression of MCT1 and then decreased the iNOS expression(P<0.01), while overexpression of MCT1 through plasmid transfection increased iNOS expression(P<0.01), while the ARG1 expressions in both conditions were not changed significantly. Conclusion After microglia polarized into inflammatory phenotype during ischemia period, iNOS production is related with MCT1 expression, and MCT1 is in the upstream of iNOS pathway.

11.
Mem. Inst. Oswaldo Cruz ; 116: e200417, 2021. tab, graf
Article in English | LILACS | ID: biblio-1154880

ABSTRACT

BACKGROUND Toxoplasma gondii causes toxoplasmosis and is controlled by activated macrophages. However, infection of macrophages by tachyzoites induces TGF-β signaling (TGF-s) inhibiting nitric oxide (NO) production. NO inhibition may be a general escape mechanism of distinct T. gondii strains. OBJECTIVES To evaluate in activated macrophages the capacity of T. gondii strains of different virulence and genetics (RH, type I; ME-49, type II; VEG, type III; P-Br, recombinant) to evade the NO microbicidal defense system and determine LC3 loading to the parasitophorous vacuole. METHODS Activated peritoneal macrophages were infected with the different T. gondii strains, NO-production was evaluated by the Griess reagent, and inducible nitric oxide synthase expression, TGF-s, and LC3 localisation assayed by immunofluorescence. FINDINGS Only RH persisted in macrophages, while VEG was more resistant than P-Br and ME-49. All strains induced TGF-s, degradation of inducible nitric oxide synthase, and NO-production inhibition from 2 to 24 h of infection, but only RH sustained these alterations for 48 h. By 24 h of infection, TGF-s lowered in macrophages infected by ME-49, and P-Br, and NO-production recovered, while VEG sustained TGF-s and NO-production inhibition longer. LC3 loading to parasitophorous vacuole was strain-dependent: higher for ME-49, P-Br and VEG, lower for RH. All strains inhibited NO-production, but only RH sustained this effect probably because it persisted in macrophages due to additional evasive mechanisms as lower LC3 loading to parasitophorous vacuole. MAIN CONCLUSIONS These results support that T. gondii can escape the NO microbicidal defense system at the initial phase of the infection, but only the virulent strain sustain this evasion mechanism.


Subject(s)
Animals , Mice , Toxoplasma/physiology , Macrophages, Peritoneal/parasitology , Nitric Oxide Synthase/metabolism , Macrophages/parasitology , Nitric Oxide/biosynthesis , Toxoplasmosis, Animal/parasitology , Macrophages/metabolism
12.
Article in Chinese | WPRIM | ID: wpr-905900

ABSTRACT

Objective:To illustrate the effect of M1/M2 polarization of macrophages on gouty arthritis models induced with monosodium urate and reveal the molecular mechanism of total saponins from Dioscoreae Nipponicae Rhizoma to treat gouty arthritis. Method:A total of 72 male SD rats were randomly divided into four groups: normal group, model group, total saponin group (160 mg·kg<sup>-1</sup>), celecoxib group (43.3 mg·kg<sup>-1</sup>), with 18 rats in each group. Gouty arthritis models were induced by injecting monosodium urate into ankle joints bilaterally. Histopathology changes of ankle joints were observed by hematoxylin-eosin(HE) staining. Immunohistochemistry method was used to detect the protein expression change of CD68, interleukin-4(IL-4), inducible nitric oxide synthase (iNOS) and transforming growth factor-<italic>β</italic><sub>1</sub>(TGF-<italic>β</italic><sub>1</sub>). Result:HE staining results showed that the inflammation of the model group was most obvious on the third day after modeling, and the disease was in the acute stage. On day 5, the inflammation was alleviated, and on day 8, the inflammation was still present but close to normal. The total saponin group and celecoxib group could improve the pathological changes of synovial tissue, and the effect of total saponin group was more obvious. Immunohistochemical results were as follows. Compared with the normal group. The expression of CD68 and iNOS in the model group increased on the 3rd,5th and 8th day of administration (<italic>P</italic><0.01). Compared with the model group, the total saponins group could reduce the expression of CD68 and iNOS (<italic>P</italic><0.05,<italic>P</italic><0.01)on the 3rd day of administration, and significantly reduced them expression on the 5th and 8th days (<italic>P</italic><0.01). Compared with the normal group, IL-4 and TGF-<italic>β</italic><sub>1</sub> expression were increased in the model group when the drug was given for three days(<italic>P</italic><0.01). Total saponin group could enhance IL-4 expression(<italic>P</italic><0.05)and decreased the TGF-<italic>β</italic><sub>1</sub> expression(<italic>P</italic><0.01). Compared with normal group, the expression of IL-4 in the model group decreased on the 5th and 8th day of administration (<italic>P</italic><0.01), and the expression of TGF-<italic>β</italic><sub>1</sub> in the model group decreased on the 5th day of administration(<italic>P</italic><0.01). Compared with the model group, the total saponins group could increase the expression of IL-4 and TGF-<italic>β</italic><sub>1</sub> at 5 d and 8 d after administration (<italic>P</italic><0.01). Conclusion:Total saponins from Dioscoreae Nipponicae Rhizoma has the potential effect to treat gouty arthritis by regulating M1/M2 polarization.

13.
Organ Transplantation ; (6): 421-2021.
Article in Chinese | WPRIM | ID: wpr-881526

ABSTRACT

Objective To evaluate the effect of interleukin (IL)-10 on donor lung function after ex vivo lung perfusion (EVLP) in rats of cardiac death. Methods Twenty adult male SD rats were randomly divided into the simple perfusion group (group A, n=10) and modified perfusion group (group B, n=10). Perfusate A (without IL-10) and perfusate B (supplemented with IL-10) was administered in group A and B, respectively. The EVLP rat models of cardiac death were established. The appearance of donor lung, dry-to-wet (D/W) ratio of donor lung tissues, the function and metabolism of donor lung, the morphology of donor lung and the levels of inflammatory markers of donor lung were statistically compared between two groups. Results After perfusion, evident edema of the whole donor lung, poor compliance and a large amount of edema fluid discharged from the airway were observed in group A, whereas no obvious edema and good compliance were found in group B. Compared with group A, the D/W ratio of lung tissues in group B was higher (P < 0.05). In both groups, the pulmonary vein partial pressure of oxygen reached the peak at 2 h after perfusion, which did not significantly differ between two groups (P > 0.05). In group B, the pulmonary artery pressure was increased at a lower speed and significantly lower after perfusion, and the lactic acid level in the perfusate was significantly lower than those in group A (all P < 0.05). In group A, the alveolar structure was largely destroyed and the cells was rare. In group B, the alveolar structure was relatively normal without evident cell edema. The incidence of cell apoptosis of donor lung was high in group A, whereas no obvious cell apoptosis of donor lung was noted in group B. After perfusion for 4 h, the levels of monocyte chemoattractant protein (MCP)-1 and IL-6 were significantly increased, the IL-4 levels were remarkably decreased (all P < 0.05), but the levels of tumor necrosis factor (TNF)-α, IL-1α and inducible nitric oxide synthase (iNOS) did not significantly change in both groups (all P > 0.05). Conclusions IL-10 may improve the function of donor lung after EVLP in rat of cardiac death by reducing cell apoptosis.

14.
Acta Anatomica Sinica ; (6): 554-560, 2021.
Article in Chinese | WPRIM | ID: wpr-1015435

ABSTRACT

Objective To explore the effects of Lycium barbarum soup on retina histological structure and related active substances in mice injured by heroin. Method Totally 120 Kunming mice were randomly divided into control group, heroin group, Lycium barbarum 1 group and Lycium barbarum 2 group. The mice were intraperitoneal

15.
Article in Chinese | WPRIM | ID: wpr-872950

ABSTRACT

Objective::To investigate the effect of betulic acid(BA) on steatosis LO2 cells. Method::LO2 cells were intervened with BA at different gradient concentrations (0, 10, 20, 40, 80, 160, 250 μmol·L-1) for 24 hours. methyl thiazolyl tetrazolium(MTT) staining was used to observed cell viability to determine the final concentration of BA. The cells were divided into control, model, dimethylsulfoxide (DMSO) and BA groups, as well as BA groups intervened with low, middle and high concentrations. First, model, DMSO and BA group's cells were cultured in 10% Lipid Mix 1 medium for 24 hours to establish a nonalcoholic fatty liver model. Then, DMSO group and low, medium and high-concentration groups were separately cultured with 0.1%DMSO medium and 20, 40, 80 μmol·L-1 BA medium for 24 hours. And control and model groups were cultured in drug-free medium for 24 hours. Oil red O staining and Nile red staining were used to observe the intracellular lipid droplets. Immunofluorescence was used to detect the protein expression of inducible nitric oxide synthase (iNOS). Western blot was used to detect the protein expression levels of receptor for advanced glycation end-products (RAGE), nuclear factor κB p65 (NF-κB p55) and iNOS. Result::BA within the concentration of 80 μmol·L-1 had no significant toxicity on LO2 cells. Compared with control group, the intracellular lipid droplets were significantly increased in the model group, and the expressions of oxidative stress-related proteins RAGE, NF-κB p65 and iNOS also increased significantly(P<0.05). Compared with model group, the intracellular lipid droplets in DMSO group were similar to those in model group, with no significant difference in the three protein expressions between the two groups. However, the intracellular lipid droplets deposition in the BA group was significantly decreased. And the expressions of RAGE, NF-κB p65 and iNOS proteins in high-concentration BA group were significantly decreased(P<0.05, P<0.01). Conclusion::BA can significantly improve the intracellular fat deposition in LO2 cells, which was probably related to the inhibition of the expressions of oxidative stress-related proteins RAGE, NF-κB p65 and iNOS.

16.
Organ Transplantation ; (6): 435-2020.
Article in Chinese | WPRIM | ID: wpr-822920

ABSTRACT

Myeloid-derived suppressor cell (MDSC) is a type of heterogeneous cell derived from bone marrow, which was first found in tumor. MDSC can inhibit the function of T cell with immunosuppressive effect. In recent years, more and more studies have shown that in the field of organ transplantation, MDSC can also regulate the host's immune function, induce specific immune tolerance, and play a protective role in transplant organs, which is expected to become a new target in clinical treatment of transplant rejection. The biological characteristics of MDSC and the mechanism of immune tolerance induced by MDSC were reviewed in this paper.

17.
Article in Chinese | WPRIM | ID: wpr-754170

ABSTRACT

Objective To investigate the effects of magnesium sulfate and magnesium L-sulfonate on the neurobehavioral response and the expression of induced nitric oxide synthase (iNOS) in neurons after acute cerebral ischemia in rats. Methods One hundred and twelve male Sprague-Dawley ( SD) rats were double-blinded randomly divided into sham-operated group, middle cerebral artery occlusion ( MCAO ) group,MgSO4 treatment group,L-MgT treatment group. Each group was further divided into 6 h,12 h and 24 h subgroups according to the different detection time points. Rat MCAO models were produced following the Longa's method. And the Longa score,limb-placing test,rotarod test,and sticky tape test were performed to evaluate the neurological damage,autonomous movement and coordinate perception in the 24 h subgroup. At the end of the experiment,2,3,5-triphenyltetrazolium chloride ( TTC) was used to evaluate the area of cerebral infarction at 24 h reperfusion. Immunohistochemical staining was employed to determine the altera-tions in iNOS expression in neurons 6 h,12 h and 24 h after reperfusion. Results In behavioral evaluation:Longa score:Normal performance was observed in sham-operated group. Compared with the MCAO group,the scores of MgSO4 treatment group(1. 71±0. 18) and L-MgT treatment group(1. 14±0. 14) were decreased (t=0. 548,3. 873,all P<0. 05),and the score of L-MgT treatment group was lower than that of the MgSO4 group(t=2. 828,P<0. 05). Limb symmetry score:There was no statistically significant difference between MgSO4 group and MCAO group,but there was a statistically significant difference between L-MgT group and MCAO group (t=7. 071,P<0. 05). The roding experiment:The time of MgSO4 group and the L-MgT group were significantly different from that of the MCAO group (t=9. 588,20. 776,P<0. 05),and the time of the L-MgT group was significantly higher than that of the MgSO4 group (t=4. 983,P<0. 05). The right limb strip removal experiment: The time of MgSO4 group and L-MgT group were statistically different from that of MCAO group (t=6. 135,5. 825,P<0. 05),and the time of L-MgT group was increased compared with that of MgSO4 group(t=4. 507,P<0. 05). TTC test:No infarction was formed in the sham group. Compared with MCAO group ((36. 82±1. 35)%),the cerebral infarction volume of MgSO4 group ((17. 39±1. 72)%) and L-MgT group ((10. 81 ± 1. 35)%) significantly decreased, with statistically significant differences ( t=8. 874,11. 105,P<0. 05). Compared with MgSO4 group,cerebral infarction volume in L-MgT group was sig-nificantly reduced,with statistical significance (t=2. 593,P<0. 05). HE staining:There was no statistically significant difference in cell morphology between MgSO4 group and MCAO group at each time point,but the cell morphology of L-MgT group was intact compared with that of MCAO group. INOS staining at 24 h:There was no statistically significant difference in the positive cell density between the MgSO4 group and the MCAO group,but the L-MgT group (cortex:(196. 7±8. 1);striatum:(153. 3±3. 8)) positive cell density was lower than that of the MCAO group (cortex:(375. 0±6. 7),striatum:(358. 3±4. 5)),and the difference was sta-tistically significant (t=11. 113,36. 231,P<0. 05). Conclusion L-MgT may have a significantly protective effect on MCAO rats,and its mechanism may be related to the level of iNOS in neurons.

18.
Article in Chinese | WPRIM | ID: wpr-742717

ABSTRACT

Objective:To observe the effect of CoCl2on the cisplatin sensitivity of human ovarian cancer SKOV3cells, and to clarify the possible mechanism.Methods:The SKOV3cells were cultured in vitro and randomly divided into control group, CoCl2 group, cisplatin (DDP) group and CoCl2 combined with DDP (combination) group.The cells in CoCl2group were cultured in normal cell medium for 20hafter cultured in 200μmol·L-1 CoCl2for 4h, the cells in DDP group were cultured in normal cell medium containing 10mg·L-1 DDP for 24h, and the cells in combination group were cultured in 10mg·L-1 DDP for 20hafter cultured in 200μmol·L-1 CoCl2for 4h.The survival rates of SKOV3cells in various groups were detected by MTT method, and the positive expression intensities of hypoxia-inducible factor-1α (HIF-1α) and inducible nitric oxide synthase (iNOS) in the cells in various groups were detected by immunofluorescence method.Rhod 2-AM fluorescence probe was used to observe the levels of Ca2+in mitochondria in the cells in various groups.Western blotting method was used to observe the expression levels of cytochrome C (cyto C) , cysteinyl aspartase 3 (caspase 3) and cleaved cysteinyl aspartase 3 (cleaved caspase 3) .Muse○R apoptosis assay kit was used to detect the apoptotic rates of cells in various groups.Results:Compared with control group, the survival rate of the cells in CoCl2group had no significant change (P>0.05) , and the survival rates of the cells in DDP and combination groups were decreased (P<0.05) ;the survival rate in combination group was higher than that in DDP group (P<0.05) .Compared with control group, the positive expression intensities of HIF-1αin CoCl2and combination groups were increased (P<0.05) .Compared with control group, the positive expressions of iNOS in DDP and combination groups were increased (P<0.05) .The Ca2+levels in the cells in DDP group and combination groups were higher than that in control group (P<0.05) and the Ca2+level in DDP group was higher than that in combination group (P<0.05) .Compared with control group, the expression levels of cyto C, caspase 3and cleaved caspase 3proteins in the SKOV3cells in CoCl2group had no significant changes (P>0.05) , and the expression levels of cyto C, caspase 3and cleaved caspase 3in DDP group were increased significantly (P<0.05) ;compared with DDP group, they were lower than those in combination group (P<0.05) .Compared with control group, the apoptotic rate of SKOV3cells in DDP group was increased significantly (P<0.05) ;the apoptotic rate of SKOV3cells in combination group was lower than that in DDP group (P<0.05) .Conclusion:CoCl2can redece the mitochondrial apoptosis of human ovarian cancer SKOV3cells by inhibiting the DDP-induced enhancement of iNOS expression and decrease the sensitivity of SKOV3cells to cisplatin.

19.
Article in Chinese | WPRIM | ID: wpr-743177

ABSTRACT

Objective To investigate the expression and significance of inducible nitric oxide synthase (iNOS), platelet-derived growth factor (PDGF)-B and lipopolysaccharide (LPS) in rat models with Budd-Chiari syndrome (BCS) . Methods BCS model was established by partial ligation of inferior vena cava in the posterior segment of the liver. The experimental rats were divided into control group (n=20), model group (n=20) and sham group (n=20) . Liver tissues were collected for immunohistochemistry, HE and Masson staining, and the expression levels of iNOS, PDGF-B and LPS were determined. Results The LPS value in model group was higher than that in both control group and sham group (P=0.001) . The mRNA and protein expressions of iNOS and PDGF-B in model group were higher than those in both control group and sham group (P=0.001) . Statistically significant differences in mRNA and protein expressions of iNOS and PDGF-B existed between each other among the subgroups (P=0.001) . In model group iNOS was positively correlated with PDGF-B and LPS; liver fibrosis was positively correlated with LPS and negatively correlated with PDGFB. Conclusion The damage and repair of BCS is a complicated process. The iNOS, PDGF-B and LPS may play different roles in different stages of BCS. How to regulate their balance in liver fibrosis may be a direction that deserves further study.

20.
Article in Chinese | WPRIM | ID: wpr-802419

ABSTRACT

Objective: To evaluate the effect of Houpu Sanwu Tang on the postoperative ileus (POI), and observe its underlying mechanisms of action on interstitial cells of cajal (ICC) and inducible nitric oxide synthase(iNOS) regulation of POI. Method: Totally 87 healthy adult male SD rats were randomly divided into sham operation group, saline control group and Houpu Sanwu Tang group at low, medium and high doses. Houpu Sanwu Tang low, middle and high dose groups received orally Houpu Sanwu Tang(2.25,4.5,9 g·kg-1); Sham operation group (Sham operation) and saline control group (Saline control) received orally normal saline. Surgical procedure was used to induce the postoperative ileus. Changes in intestinal propulsion rate, intestinal mucosal injury and small intestine expression of c-kit and iNOS among these groups were detected. Result: Intestinal propulsion rate was significantly higher in Houpo Sanwu Tang group than that in Saline control group (PPPPConclusion: Houpu Sanwu Tang can improve the intestinal propulsion rate and the recovery in POI rats. The mechanisms are related to the inhibition of the generation of iNOS, the alleviation of inflammatory response, and increase of the number of ICC, so as to ensure its normal function, and improve the intestinal dynamic disorder.

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